Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Response to Amendment
The Amendment filed 12/03/2025 has been entered. Claims 1-2 and 6-24 are pending in the application. Claims 3-5 were previously cancelled. Claims 1-2, 9, 11-12, 15, 18, 21-22 are currently amended and claims 23-24 are newly added. Applicant states in the remarks that claims 23-27 are newly added. However, claims 25-27 do not exist in the newly amended claims. Therefore, only claims 1-2 and 6-24 will be examined herein.
Status of Objections and Rejections
The objection to claims 12 and 21 from the previous office action is withdrawn in view of Applicant's amendment. The objection to claim 22 is withdrawn based on dependency of all of the limitations of claim 21.
The rejection of claim 2 under 35 U.S.C. 112(b) from the previous office action is withdrawn in view of Applicant's amendment.
The rejection of claims 1-2, 6-7, 12-15, 18-20 under 35 U.S.C. 103 from the previous office action is maintained. Claims 8, 9-11, 16-17, 21-23 22 are rejected based on dependency of all of the limitations of claims 1, 15, and 21, respectively.
New grounds of claim objections are necessitated by the amendments.
New grounds of rejection under 35 U.S.C. 112(b) are necessitated by the amendments.
New grounds of rejection under 35 U.S.C. 103 are necessitated by the amendments.
Response to Arguments
Applicant’s arguments, see pages 9-17, filed 12/03/2025, with respect to the rejection of claims 1-2, 6-7, 12-15, and 18-20 under 35 U.S.C. as being unpatentable over Menon et al. (US 20140273058 A1) with Pall Life Sciences (“Vivid™ Plasma Separation Membrane; 2009) incorporated by reference, in view of Marble (US 20130184446 A1), and in further view of Lee et al. (US 20150090674 A1) and Miki et al. (US 20170258977 A1), have been fully considered but they are not persuasive.
Applicant argues (pp. 10-11) that Menon uses adsorption (e.g. affinity-based filters) as opposed to “pore-size/structure based filters” as is therefore an unfit analogous art.
The Examiner respectfully disagrees.
In response to applicant's argument that Menon is nonanalogous art, it has been held that a prior art reference must either be in the field of the inventor’s endeavor or, if not, then be reasonably pertinent to the particular problem with which the inventor was concerned, in order to be relied upon as a basis for rejection of the claimed invention. See In re Oetiker, 977 F.2d 1443, 24 USPQ2d 1443 (Fed. Cir. 1992).
Even though Menon emphasizes adsorption, it still uses membranes that include a porous structure demonstrated to retain the desired cells and allow non-target cells to pass through. For example, Menon’s asymmetric membrane has varying pore size (large to small) across the flow direction of the sample that retains larger blood cell components upstream ([0077]). Menon is therefore analogous art since it is in the same field of endeavor as the claimed multi-stage porous filtration system.
Applicant argues (p. 11) that although Marble seems to be a pore-size based multi-layered filter system, it is an unfit analogous prior art reference as well. Marble aims to purify microorganism nucleic acid from a whole blood sample as opposed to retaining and collecting different blood components at each of the multiple filter units in the claimed invention.
The Examiner respectfully disagrees.
In response to applicant's argument that Menon is nonanalogous art, it has been held that a prior art reference must either be in the field of the inventor’s endeavor or, if not, then be reasonably pertinent to the particular problem with which the inventor was concerned, in order to be relied upon as a basis for rejection of the claimed invention. See In re Oetiker, 977 F.2d 1443, 24 USPQ2d 1443 (Fed. Cir. 1992).
Although Marble aims to purify microorganism nucleic acid from a whole blood sample, it also teaches the structure of a multi-stage column used “to accomplish the dual aims of retaining human cells in the upper stage(s) and lysing bacterial/fungal cells and viruses in the lower stage” ([0049]). Paragraph [0015] lists all of the different cell components that are collected at each stage using size exclusion filters (e.g., lymphocytes, monocytes, neutrophils, eosinophils, and basophils, platelets, DNA). Marble is therefore analogous art since it is in the same field of endeavor as the claimed multi-stage porous filtration system.
Applicant argues (pp. 11-12) that Marble’s first filter with a pore size of 5-15µm ([0022]) does not separate and retain all blood cells. Applicant also argues that Marble’s teachings are contradictory in stating that lymphocytes both pass through and are retained at the first filter ([0014]).
In response to applicant's argument that the references fail to show certain features of the invention, it is noted that the features upon which applicant relies (i.e., the first filter retains all blood cells) are not recited in the rejected claim(s). Although the claims are interpreted in light of the specification, limitations from the specification are not read into the claims. See In re Van Geuns, 988 F.2d 1181, 26 USPQ2d 1057 (Fed. Cir. 1993).
Applicant’s argument that Marble fails to retain all blood cells is not persuasive because the claims do not require this limitation. Marble’s teaching of both retaining and allowing the passage of lymphocytes does not render the reference inoperative. Different operating conditions can be accounted for in this statement as well as subpopulations of lymphocytes (e.g., B cells, T cells) which have different sizes. Additionally, Marble teaches a filter size of 10µm which is within the claimed ranged of 5-15µm which is sufficient to meet the structural limitation and is therefore capable of performing the claimed function of “separating and retaining a first set of blood components from the whole blood sample, letting through a first passing-through stream, wherein the first set of blood components include blood cells” (claims 1 and 15). Differences in the degree of separation (all blood cells) reflect routine optimization of a result-effect variable, pore size, and do not distinguish over the prior art (See MPEP 2144.05(II)).
Applicant argues (p. 12) that the prior art reference, Lee, discloses ranges for thickness and basis weight but does not specify which layer those ranges apply to. Therefore, if a person skilled in the art adopts ranges of the filter member disclosed in Lee, this range would need to be applied to all the filter units rather than only the first. As a result, applying those ranges to the first filter unit of the claim is based on hindsight. Applicant also notes that the multi-layered filter system of Lee is for collecting serum and plasma, an aim that is different from the claimed invention.
In response to applicant's argument that the examiner's conclusion of obviousness is based upon improper hindsight reasoning, it must be recognized that any judgment on obviousness is in a sense necessarily a reconstruction based upon hindsight reasoning. But so long as it takes into account only knowledge which was within the level of ordinary skill at the time the claimed invention was made, and does not include knowledge gleaned only from the applicant's disclosure, such a reconstruction is proper. See In re McLaughlin, 443 F.2d 1392, 170 USPQ 209 (CCPA 1971).
Lee teaches filter membranes in a multi-layer system having thickness and basis weight ranges that fall within the claimed ranges which are relevant to the claimed invention. A person of ordinary skill in the art would have understood such parameters to be applicable to individual filter layers. The rejection does not rely on Lee specifying the exact layer corresponding to the claimed first filter unit, but rather on Lee’s disclosure of filter layers having the claimed structural characteristics. Applying known ranges to a particular layer in a multilayer system constitutes no more than predictable use of prior art elements according to their established functions and does not involve hindsight (See MPEP 2141(I)).
In response to applicant's argument that Lee is nonanalogous art, it has been held that a prior art reference must either be in the field of the inventor’s endeavor or, if not, then be reasonably pertinent to the particular problem with which the inventor was concerned, in order to be relied upon as a basis for rejection of the claimed invention. See In re Oetiker, 977 F.2d 1443, 24 USPQ2d 1443 (Fed. Cir. 1992).
In this case, a person of ordinary skill in the art would have reasonably relied on Lee for these structural teachings regardless of its stated purpose, as references from analogous fields addressing similar structural teachings or problems are properly combinable. The difference in intended uses does not negate the applicability of Lee’s teaching. Regardless, the apparatus of Lee is used for separating plasma or serum from blood, which is consistent with that of the instant applicant since serum is a blood component (Lee, Abstract).
Applicant argues (p. 12-13) that the rejection improperly combines Marble and Lee by taking pore size (5-5µm) from Marble and taking thickness (500-1500 µm) from Lee. Lee’s disclosure ties thickness (0.8-1.2 mm) to pore size (1.25-2.75 µm). Therefore, selecting Lee’s thickness while ignoring its pore size is “cherry picking” and hindsight. The Applicant further argues that a person of ordinary skill in the art would not adopt one parameter from Lee while disregarding the rest.
In response to applicant's argument that selecting Lee’s thickness while ignoring its pore size is “cherry picking”, the test for obviousness is not whether the features of a secondary reference may be bodily incorporated into the structure of the primary reference; nor is it that the claimed invention must be expressly suggested in any one or all of the references. Rather, the test is what the combined teachings of the references would have suggested to those of ordinary skill in the art. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981).
Thickness and pore size are independent result-effective variables and Lee does not require that its thickness and pore size be used together exclusively. Lee even states that filters of different pore sizes can be used and that filters can even be stacked atop one another to meet thickness conditions ([0083]). As such, pore size and thickness are separate structural parameters that may be independently selected. A person of ordinary skill in the art would have recognized that the first filter stage is most susceptible to clogging and mechanical stress due to the initial load and concentration of the sample and would have been motivated to provide a greater thickness and larger pore size at the upstream filter to increase capacity and durability. Thickness is optimized for mechanical properties and pore size for separation characteristics. Selecting one does not require adopting the other. The rejection does not ignore Lee’s teachings but instead selects relevant features for their known purposes which is permitted use of prior art and not hindsight. Combining the two would yield predictable filtration performance especially in that Lee does not discourage from using different pore sizes and even discloses more than one embodiment. Selecting pore size from one reference and thickness from another is a matter of routine optimization of known parameters (See MPEP 2144.05).
Applicant argues (p. 13) that the rejection improperly combines Menon, Marble, and Lee by selecting isolated features from each reference, the references are loosely related with different goals, and there is no articulated reason to combine them. Therefore, the combination is “cherry picking” and based on hindsight.
The Examiner respectfully disagrees.
Menon, Marble, and Lee teach a multi-layer detachable blood filtration system, with the system of Menon being detachable; The cited references are all therefore analogous art. The rejection does not rely on arbitrary selection of features, but rather applies each reference for its respective teaching, including Menon for a multi-layer detachable blood filtration system, Marble for pore size selection, and Lee for filter thickness. The office action (pp. 8-12) clearly explains reasons for combining the different filter parameters from each reference. The intention of Menon is to first filter cells, and then proteins to avoid degradation of the nucleic acids and is relied upon to teach a second, third and fourth filter unit stacked atop each other. Reference Marble is relied upon to teach the addition of the first filter with a pore size greater than Menon’s second filter in order to reduce clogging downstream which is supporting by Menon ([0085]). Menon is silent to teaching the pore size of the third filter so Marble is sought to teach a pore size being within a range that is smaller than that of the previous filter, since the filtered components would naturally decrease in size after each layer. Reference Lee is used to cure the deficiency of the thickness and weight of the first filter by suggesting that filter elements may be substituted and layered to achieve desired properties, further supporting the combination ([0083]). A person of ordinary skill in the art would have been motivated to combine these teachings to achieve predictable filtration performance, as selection of pore size and thickness are well-known result-effective variables in filtration systems that can be optimized (See MPEP 2144.05). Accordingly, the rejection is based on the predictable use of prior art elements according to their established functions, and does not rely on impermissible hindsight (See MPEP 2141(I)).
Applicant argues (p. 14) that the cited combination of Menon, Marble, and Lee and Miki fails to provide guidance on which feature applies to which layer and that the rejection ignores a key feature of the claimed invention which is a particular order of distinctive filter units.
The Examiner respectfully disagrees.
The ordering of filter units in a multi-layer filtration system would have been understood by a person of ordinary skill in the art to be dictated by functional considerations, such as progressively removing larger to smaller particles. The cited references collectively teach multi-layer filtration systems and filter characteristics (e.g., pore size, thickness), and a person of ordinary skill in the art would have arranged such layers in a predictable order to achieve effective filtration. The references need not explicitly identify each layer as “first” or “second,” as the arrangement follows from well-known filtration principles and represents a matter of routine optimization. Accordingly, the ordering limitation does not distinguish the claimed invention from the prior art.
Claim Objections
Claims 1-2 and 6-24 are objected to because of the following informalities:
Regarding claim 1, lines 7, 11, and 14 state “include”. Applicant can amend the instant claim to recite “includes” since the subject “set” is singular.
Claims 2, 6-14, and 23 are objected to based on dependency of all of the limitations of claim 1.
Regarding claim 15, lines 8, 13, and 18 state “include”. Applicant can amend the instant claim to recite “includes” since the subject “set” is singular.
Claims 16-20 and 24 are objected to based on dependency of all of the limitations of claim 15.
Regarding claim 21, ll. 30 state “include”. Applicant can amend the instant claim to recite “includes” since the subject “stream” is singular.
Claim 21 is objected to based on dependency of all of the limitations of claim 21.
Regarding claim 23, lines 3 and 14 state “include”. Applicant can amend the instant claim to recite “includes” since the subject “set” is singular.
Regarding claim 24, lines 4 and 13 state “include”. Applicant can amend the instant claim to recite “includes” since the subjects “set” and “stream” are singular.
Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 21 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Regarding claim 21, line 36 recites “the affinity-facilitating material of the fourth filter unit is a silica material”. There is insufficient antecedent basis for this limitation in the claim. Applicant can correct this by stating “the fourth filter unit is made of an affinity-facilitating silica material”.
Claim 22 is rejected based upon dependency of all of the limitations of claim 21.
Claim Interpretation
The claims contain limitations which are directed to intended uses or capabilities of the claimed invention. These limitations are only given patentable weight to the extent which effects the structure of the claimed invention. Please see MPEP 2114. Note that functional limitations are emphasized in italics herein.
Claim Rejections - 35 USC § 103
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-2, 6-7, 12-15, 18-20 are rejected under 35 U.S.C. 103 as being unpatentable over Menon et al. (US 20140273058 A1) with Pall Life Sciences (“Vivid™ Plasma Separation Membrane; 2009) incorporated by reference, in view of Marble (US 20130184446 A1), and in further view of Lee et al. (US 20150090674 A1) and Miki et al. (US 20170258977 A1).
Regarding claim 1, Menon teaches a multi-layered filter (an integrated membrane assembly 30; [0087]; Fig. 2) for filtering and retaining a plurality of blood components of a whole blood sample when passing through a plurality of sequentially stacked filter units that constitute the filter (for separation and preservation of biomolecules. Classes of biomolecules, for example, those in blood, can be separated and preserved; Abstract)(The top side 20 is exposed to whole blood while the bottom side 22 is exposed to the flow-through processed sample; [0075]; Fig. 1)(the layers 12, 14, 16 are stacked and pressed together as a single unit ; [0085]; Fig. 1; filters 32, 34, and 36 of Fig. 2) comprising:
a second filter unit (a sample collection layer 32 (alternatively referred to as a cell collection layer; [0087]; Fig. 2), having a pore size of 0.05-1 um (“The sample collection layer 32 comprises an asymmetric membrane,” wherein the “asymmetric membrane [has a]…pore size of 0.2 .mu.m.; [0087][0082]; Fig. 3) separating and retaining a second set of blood components, letting through a second passing-through stream, wherein the second set of blood components include blood platelet or exosome (“The porosity of the membrane was carefully selected in order to trap and remove cell/cellular debris while allowing nucleic acid species and proteins to flow through,” wherein cell is defined to include “platelets”; [0100][0069]), having a thickness of 100-1000 pm, and a weight of 100-300 gram/m2 (Paragraph [0077] of Menon states the “asymmetric membrane is a VIVID Plasma Separation Membrane (Pall Corporation, Port Washington, N.Y.)” which has a thickness of 330 um according to the specification on page 1 of Pall’s product data sheet (see attached document). Additionally, a weight of 161 gram/m2 can be calculated using this thickness, the 1.24 g/cm3 density of polysulfone, and the void volume of 20 ul/cm2 of the membrane (see page 2, column 1, paragraph 2, lines 7-8 of the attached product data sheet for Pall Life sciences),
a third filter unit (“a second layer for protein adsorption,” or “a protein adsorption layer 34”; Abstract; [0087]; Fig. 2) separating and retaining a third set of blood components, letting through a third passing-through stream from the second passing-through stream, wherein the third set of blood components include proteins (a protein adsorption layer 84 designed to selectively remove proteins from the cell free flow through, wherein the third passing-through stream contained nucleic acid species; [0100]); and
wherein the second and third filter units are sequentially stacked in a particular sequential order (sequence the filtering process to first filter cells and then proteins to avoid degradation of the nucleic acids; [0026]; See order of layers 32, 34, and 36 of Figs. 2-3) in which the third filter unit is behind the second filter unit, in the direction of the whole blood sample passing through the plurality of sequentially stacked filter units (the layers 12, 14, 16 are stacked and pressed together as a single unit ; [0085]; Fig. 1; filters 32, 34, and 36 of Fig. 2)(The top side 20 is exposed to whole blood while the bottom side 22 is exposed to the flow-through processed sample; [0075]; Fig. 1), detachably connected to each other (each holder 64 is capable of removably engaging at least one adjacent holder 64; [0091]; Figs. 2-3) with predetermined spaces separating therebetween (Paragraph [0091] explains that each layer 32, 34, and 36 is encased within its own holder 64 which has an upper and lower portion 66 and 68, wherein the lower portion of one holder snaps into the upper portion of the holder beneath it. Fig. 3 shows this housing to have a circular opening on the upper and lower portions revealing the filters (e.g. 36) in between which naturally creates a void space and is predetermined in that the reference chose to encase the filters)(See spacing being layers in Fig. 8), and disassembled after filtering (This pie-shaped design of the membrane 32, 34, 36 allows a user to easily separate sections of each of the membrane 32, 34, 36 for easy partial sample analysis; [0092]; Fig. 3) and wherein after the plurality of sequentially stacked filter units are disassembled after filtering the whole blood sample, the second set of blood components retained at the second filter unit are collected for a second further test or experiment pertaining to the second set of blood components, and a third set of blood components retained at the third filter unit are collected for a third further test or experiment pertaining to the third set of blood components (The multi-layered filter taught by Menon is functionally capable of performing these analysis steps since the each of the layers can be disassembled for partial sample analysis ([0092]; Fig. 3)).
Menon fails to teach:
a first filter unit, having a pore size of 5-15 um, a thickness of 500-1500 um, and a weight of 100-400 gram/m2, separating blood cells from a whole blood sample
a third filter unit having a pore size of 0.003-0.01 um (emphasis added), and
the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes.
Menon, however, does teach that all of the filters have a porous structure ([0088]), and instead of an acrylic plate, Menon shows the stacked filters encased above a “bottom portion 50” ([0087; Figs. 2-3).
Marble teaches:
a first filter unit, having a pore size of 5-15 um, (the first size-exclusion filter has a pore size of about 10 µm; [0022]), separating blood cells (excludes passage therethrough of lymphocytes, monocytes, neutrophils, eosinophils, and basophils; [0015]) from a whole blood sample (the sample comprises whole blood; [0013])
a third filter unit having a pore size of 0.01 um (“the shearing filter has a pore size of about 0.01-0.4 µm; [0015]).
Marble is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon to incorporate the teachings of Marble by adding a first filter unitabove the second filter unit, having a pore size of 5-15 um, separating blood cells from a whole blood sample, in order to reduce clogging in the subsequent layers. Primary reference, Menon, teaches the second layer to be an asymmetric membrane comprising larger pores on an upstream portion and smaller pores on a downstream portion, where the larger upstream pores act as a pre-filter while the smaller downstream pores serve as an absolute cut-off layer ([0085]). Menon explains that this structure efficiently captures cells, including erythrocytes and platelets, while reducing clogging ([0085])(emphasis added). One of ordinary skill in the art would have been motivated to include a first, cell-separating filter above the asymmetric membrane in order to remove bulk whole blood cells early, protect finer downstream filters from fouling, and improve recovery of smaller analytes such as proteins and nucleic acids. The motivation is consistent with MPEP 2143(I)(A), as this is merely combining prior-art elements according to known methods to yield predictable results since both the first filter of Marble and the multilayer filter of Menon would each perform the same functions as they do separately.
Modified Menon fails to teach the first filter, having a thickness of 500-1500 um, and a weight of 100-400 gram/m2 , and the third filter unit having a pore size of 0.003-0.01 um (emphasis added), the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes.
Lee teaches a filter having a thickness of 500-1500 um (“The filter member may have a thickness in a range from about 0.8 to about 1.2 mm,” wherein this equates to a range of 800-1200um; [0029]), and a weight of 100-400 gram/m2 (The filter member may have a weight per surface area (or a basis weight) in a range from about…70 to about 120 g/m.sup.2; [0029]).
Lee is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon in view of Marble to incorporate the teachings of Lee by adding a first filter, having a thickness of 500-1500 um and a weight of 100-400 gram/m2, in order to “to reduce the occurrence of hemolysis of corpuscles including erythrocytes” (Lee, [0028]). Primary reference, Menon expresses a desire to collect blood cells without lysis ([0101]). Secondary reference Marble teaches staged size-exclusion filters with the first filter dimensioned for whole-cell capture ([0015]). Lee teaches that blood filter members having a thickness and weight per surface area in a particular range provide high plasma recovery while preventing hemolysis ([0028][0029]). It would have been advantageous to select Lee’s disclosed thickness and weight for the first filter of Menon in view of Marble to ensure durability, capacity, and reduced cell lysis, as both Lee and Menon stress the importance of avoiding red blood cell rupture. The overlapping filter weight range between the teaching of Lee and the instant claim is a matter of optimizing a routine result-effective variable to yield the predictable outcome of a lower amount of lysed cells (See MPEP 2144.05). It would have been obvious before the effective filing date of the claimed invention to have chosen these filter characteristics so as to achieve such a benefit.
Modified Menon fails to teach the third filter unit having a pore size of 0.003-0.01 um (emphasis added), the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes.
Modified Menon instead teaches that the third filter has a pore size of about 0.01 μm which is a partial overlap (Marble, [0015]) of the claimed range. Primary reference, Menon, teaches a protein-removal layer in a stacked filter assembly (See Fig. 2) and explicitly states that each layer is a porous membrane ([0088]) used to deplete proteins so nucleic acids are preserved for downstream analysis (Abstract). One of ordinary skill in the art seeking to ensure protein separation (the function of the third filter as stated in the abstract of the instant publication US20220168672A1) would naturally look to known pore-size implementations for fine cutoffs. Marble discloses a downstream filter stage with pores as small as about 0.01 um within the same kind of multi-stage, blood sample filtration stack, demonstrating the feasibility and predictability of using micro to sub-micron cutoffs in this context. Pore size is a result-effective variable governing what passes or what is retained. Because Marble touches the claimed band at 0.01 um, modifying the filter taught by Menon by selecting a 0.01 um filter, and then routinely optimizing this result-effective variable to slightly below (0.003-0.01) would have been obvious to one having an ordinary skill in the art before the effective filing date of the invention (MPEP 2144.05(I)).
Modified Menon fails to teach the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes in decreasing order.
Miki teaches filter layers stacked (see filter layers above 61 in Fig. 2) on an acrylic plate (second end plate 61; [0074]).
Miki is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon in view of Marble and Lee to incorporate the teachings of Miki by stacking the filter layers on an acrylic plate so as to provide additional rigid support using a lightweight material that is easily processed (Miki, [0074]). Substitution of the bottom portion 50 taught by Menon with the acrylic second endplate 61 of Miki would yield predictable results since they both serve the same purpose for providing a steady support for the device (See MPEP 2143(I)(B)).
Modified Menon fails to teach each of the filter units has different pore sizes.
Marble teaches that each filter unit has a different pore size (paragraphs [0006]-[0007] list pore size ranges that do not overlap for three different filters).
Marble is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon in view of Marble, Lee, and Miki to further incorporate the teachings of Marble by having filter units of different pore sizes “such that fluid passing from the proximal end to the distal end of the sample containment vessel passes through the first, second, and third filters” (Marble, [0006]). One of ordinary skill in the art before the effective filing date of the claimed invention would have recognized that staged, decreasing pore size is a routine and predictable design in multi-layer filters that would yield the predictable results of efficient fractionation of blood components (MPEP 2143(I)(B)).
Regarding claim 2, Modified Menon teaches the multi-layered filter according to claim 1, wherein the whole blood sample passed through the third filter unit includes at least one of amino acids, bases and various metabolites (the multi-layer filter taught by modified Menon is functionally capable of passing of amino acids, bases or various [metabolites] through the third filter. Menon states that the hydrophobic C18 filter of the protein adsorption layer allows nucleic acids to remain in the sample flow and are stabilized downstream in the fourth layer ([0079]). Molecules smaller than nucleic acids lacking the hydrophobic portions capable of interacting with the C18, such as those mentioned in the claim limitation, would naturally pass through the third filter)(See 112(b) rejection above for the interpretation of “various” to mean “various metabolites”).
Regarding claim 6, Modified Menon teaches the multi-layered filter according to claim 1, wherein the multi- layered filter is configured to separate the whole blood sampled administrated in an amount of 20- 100 uL (sample volumes as small as 5 .mu.l up to 150 .mu.l of whole blood.; [0106]).
Although modified Menon teaches a slightly broader range than the instant claim, the selection of a more narrow whole blood sample in an amount 20-100 ul would have been a matter of routine optimization of a result-effective variable (See MPEP 2144.05(I)). It would have been obvious before the effective filing date of the claimed invention to have chosen this range of whole blood volume so as to yield the predictable outcome of a better separation of blood sample components.
Regarding claim 7, Menon teaches a disease diagnosis kit (an integrated membrane assembly 30; [0087]; Fig. 2) configured to measure blood P-selectin by using the multi-layered filter according to claim 1 (The multi-layered filter taught by modified Menon is functionally capable of measuring blood P-selectin since the protein adsorption layer of Menon has a pore size small enough to filter proteins and is highly hydrophobic which can bind to hydrophobic P-selectin. Subsequent measuring can ensue by further observation).
Regarding claim 12, Modified Menon teaches the multi-layered filter of claim 1.
Modified Menon fails to teach the third filter unit further includes additional filter unit according to a size of the protein to be separated and retained. However primary reference, Menon, teaches the second layer to be an asymmetric membrane comprising larger pores on an upstream portion and smaller pores on a downstream portion, where the larger upstream pores act as a pre-filter while the smaller downstream pores serve as an absolute cut-off layer ([0085]). It would have been obvious for one of ordinary skill in the art before the effective filing date of the invention to have substituted the membrane used in the protein adsorption layer with an asymmetric membrane for the expectation of predictable results in order to reduce clogging in the subsequent layers, a technique that is well-known in the art ([0085]) (See MPEP 2143(I)(B)).
Regarding claim 13, Modified Menon teaches the multi-layered filter of claim 1, wherein the first filter unit is made of a material selected from a group of natural fibers (Any suitable size-exclusion filter can be used…an example…cellulose acetate size exclusion filter; Marble, [0038]).
Regarding claim 14, Modified Menon teaches the multi-layered filter of claim 1, wherein the third filter unit is made of a material with low affinity and adsorption for nucleic acids (The protein adsorption layer 14 comprises a membrane conjugated to an 18-mer hydrocarbon chain (C18)… enables hydrophilic molecules, such as RNA/DNA, to flow through the layer 14 for subsequent preservation of protein-free nucleic acids; Menon, [0083]) or proteins .
Regarding claim 15, Menon teaches a method of separating a plurality of blood components from a whole blood sample by passing the whole blood sample through a plurality of sequentially stacked filter units (separate of classes of biomolecules such as cells, proteins and nucleic acids (DNA/RNA) fully preserved from whole blood; [0019])(The top side 20 is exposed to whole blood while the bottom side 22 is exposed to the flow-through processed sample; [0075]; Fig. 1)(the layers 12, 14, 16 are stacked and pressed together as a single unit ; [0085]; Fig. 1; filters 32, 34, and 36 of Fig. 2) that constitute a multi-layered filter (an integrated membrane assembly 30; [0087]; Fig. 2), the method comprising:
separating and retaining a second set of blood components through a second filter unit of the plurality of sequentially stacked filter units, letting through a second passing-through stream, wherein the second filter unit has a pore size of 0.05-1 pm (“The sample collection layer 32 comprises an asymmetric membrane,” wherein the “asymmetric membrane [has a]…pore size of 0.2 .mu.m.; [0087][0082]; Fig. 3), a thickness of 100-1000 pm, and a weight of 100-300 gram/m (Menon states the “asymmetric membrane is a VIVID Plasma Separation Membrane (Pall Corporation, Port Washington, N.Y.)” which has a thickness of 330 um according to the specification on page 1 of Pall’s product data sheet (see attached document). Additionally, a weight of 161 gram/m2 can be calculated using this thickness, the 1.24 g/cm3 density of polysulfone, and the void volume of 20 ul/cm2 of the membrane (see page 2, column 1, paragraph 2, lines 7-8 of the attached product data sheet for Pall Life sciences), and wherein the second set of blood components include blood platelet or exosome (“The porosity of the membrane was carefully selected in order to trap and remove cell/cellular debris while allowing nucleic acid species and proteins to flow through,” wherein cell is defined to include “platelets”; [0100][0069]), and
separating and retaining a third set of blood components from the second passing-through stream through a third filter unit of the plurality of sequentially stacked filter units, letting through a third passing-through stream, and wherein the third set of blood components include proteins, (a protein adsorption layer 84 designed to selectively remove proteins from the cell free flow through, wherein the third passing-through stream contained nucleic acid species; [0100]); and
wherein the second and third filter units are sequentially stacked in a particular sequential order in which the third filter unit is behind the second filter unit, in a direction of the whole blood sample passing through the plurality of sequentially stacked filter units (sequence the filtering process to first filter cells and then proteins to avoid degradation of the nucleic acids; [0026]; See order of layers 32, 34, and 36 of Figs. 2-3)(the layers 12, 14, 16 are stacked and pressed together as a single unit ; [0085]; Fig. 1; filters 32, 34, and 36 of Fig. 2)(The top side 20 is exposed to whole blood while the bottom side 22 is exposed to the flow-through processed sample; [0075]; Fig. 1), detachably connected to each other (each holder 64 is capable of removably engaging at least one adjacent holder 64; [0091]; Figs. 2-3) with predetermined spaces separating therebetween (Paragraph [0091] explains that each layer 32, 34, and 36 is encased within its own holder 64 which has an upper and lower portion 66 and 68, wherein the lower portion of one holder snaps into the upper portion of the holder beneath it. Fig. 3 shows this housing to have a circular opening on the upper and lower portions revealing the filters (e.g. 36) in between which naturally creates a void space and is predetermined in that the reference chose to encase the filters)(See spacing being layers in Fig. 8), and disassembled after filtering (This pie-shaped design of the membrane 32, 34, 36 allows a user to easily separate sections of each of the membrane 32, 34, 36 for easy partial sample analysis; [0092]; Fig. 3) and
wherein after the plurality of sequentially stacked filter units are disassembled after filtering the whole blood sample, the second set of blood components retained at the second filter unit are collected for a second further test or experiment pertaining to the second set of blood components, and the third set of blood components retained at the third filter unit are collected for a third further test or experiment pertaining to the third set of blood components (See the results of total protein concentration and RNA post-loading in Figs. 9-12)(This pie-shaped design of the membrane 32, 34, 36 allows a user to easily separate sections of each of the membrane 32, 34, 36 for easy partial sample analysis, leaving other sections of the membrane available for analysis at a later time or for concurrent analysis; [0092]).
Menon fails to teach:
a first filter unit, having a pore size of 5-15 um, a thickness of 500-1500 um, and a weight of 100-400 gram/m2, separating blood cells from a whole blood sample
a third filter unit having a pore size of 0.003-0.01 um (emphasis added), and
the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes in decreasing order.
Menon, however, does teach that all of the filters have a porous structure ([0088]), and instead of an acrylic plate, Menon shows the stacked filters encased above a “bottom portion 50” ([0087; Figs. 2-3).
Marble teaches:
a first filter unit, having a pore size of 5-15 um, (the first size-exclusion filter has a pore size of about 10 µm; [0022]), separating blood cells (excludes passage therethrough of lymphocytes, monocytes, neutrophils, eosinophils, and basophils; [0015]) from a whole blood sample (the sample comprises whole blood; [0013])
a third filter unit having a pore size of 0.01 um (“the shearing filter has a pore size of about 0.01-0.4 µm; [0015]).
Marble is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon to incorporate the teachings of Marble by adding a first filter above the second filter, having a pore size of 5-15 um, separating blood cells from a whole blood sample, in order to reduce clogging in the subsequent layers. Primary reference, Menon, teaches the second layer to be an asymmetric membrane comprising larger pores on an upstream portion and smaller pores on a downstream portion, where the larger upstream pores act as a pre-filter while the smaller downstream pores serve as an absolute cut-off layer ([0085]). Menon explains that this structure efficiently captures cells, including erythrocytes and platelets, while reducing clogging ([0085])(emphasis added). One of ordinary skill in the art would have been motivated to include a first, cell-separating filter above the asymmetric membrane in order to remove bulk whole blood cells early, protect finer downstream filters from fouling, and improve recovery of smaller analytes such as proteins and nucleic acids. The motivation is consistent with MPEP 2143(I)(A), as this is merely combining prior-art elements according to known methods to yield predictable results since both the first filter of Marble and the multilayer filter of Menon would each perform the same functions as they do separately.
Modified Menon fails to teach the first filter, having a thickness of 500-1500 um, and a weight of 100-400 gram/m2 , the third filter unit having a pore size of 0.003-0.01 um (emphasis added), the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes.
Lee teaches a filter having a thickness of 500-1500 um (“The filter member may have a thickness in a range from about 0.8 to about 1.2 mm,” wherein this equates to a range of 800-1200um; [0029]), and a weight of 100-400 gram/m2 (The filter member may have a weight per surface area (or a basis weight) in a range from about…70 to about 120 g/m.sup.2; [0029]).
Lee is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon in view of Marble to incorporate the teachings of Lee by adding a first filter, having a thickness of 500-1500 um and a weight of 100-400 gram/m2, in order to “to reduce the occurrence of hemolysis of corpuscles including erythrocytes” (Lee, [0028]). Primary reference, Menon expresses a desire to collect blood cells without lysis ([0101]). Secondary reference Marble teaches staged size-exclusion filters with the first filter dimensioned for whole-cell capture ([0015]). Lee teaches that blood filter members having a thickness and weight per surface area in a particular range provide high plasma recovery while preventing hemolysis ([0028][0029]). It would have been advantageous to select Lee’s disclosed thickness and weight for the first filter of Menon in view of Marble to ensure durability, capacity, and reduced cell lysis, as both Lee and Menon stress the importance of avoiding red blood cell rupture. The overlapping filter weight range between the teaching of Lee and the instant claim is a matter of optimizing a routine result-effective variable to yield the predictable outcome of a lower amount of lysed cells (See MPEP 2144.05). It would have been obvious before the effective filing date of the claimed invention to have chosen these filter characteristics so as to achieve such a benefit.
Modified Menon fails to teach the third filter unit having a pore size of 0.003-0.01 um (emphasis added), the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes.
Modified Menon instead teaches that the third filter has a pore size of about 0.01 μm which is a partial overlap (Marble, [0015]) of the claimed range. Primary reference, Menon, teaches a protein-removal layer in a stacked filter assembly (See Fig. 2) and explicitly states that each layer is a porous membrane ([0088]) used to deplete proteins so nucleic acids are preserved for downstream analysis (Abstract). One of ordinary skill in the art seeking to ensure protein separation (the function of the third filter as stated in the abstract of the instant publication US20220168672A1) would naturally look to known pore-size implementations for fine cutoffs. Marble discloses a downstream filter stage with pores as small as about 0.01 um within the same kind of multi-stage, blood sample filtration stack, demonstrating the feasibility and predictability of using micro to sub-micron cutoffs in this context. Pore size is a result-effective variable governing what passes or what is retained. Because Marble touches the claimed band at 0.01 um, modifying the filter taught by Menon by selecting a 0.01 um filter, and then routinely optimizing this result-effective variable to slightly below (0.003-0.01) would have been obvious to one having an ordinary skill in the art before the effective filing date of the invention (MPEP 2144.05(I)).
Modified Menon fails to teach the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes.
Miki teaches filter layers stacked (see filter layers above 61 in Fig. 2) on an acrylic plate (second end plate 61; [0074]).
Miki is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon in view of Marble and Lee to incorporate the teachings of Miki by stacking the filter layers on an acrylic plate so as to provide additional rigid support using a lightweight material that is easily processed (Miki, [0074]). Substitution of the bottom portion 50 taught by Menon with the acrylic second endplate 61 of Miki would yield predictable results since they both serve the same purpose for providing a steady support for the device (See MPEP 2143(I)(B)).
Modified Menon fails to teach that each of the filter units has a different pore size.
Marble teaches that each filter unit has a different pore size (paragraphs [0006]-[0007] list pore size ranges that do not overlap for three different filters).
Marble is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon in view of Marble, Lee, and Miki to further incorporate the teachings of Marble by having filter units of different pore sizes “such that fluid passing from the proximal end to the distal end of the sample containment vessel passes through the first, second, and third filters” (Marble, [0006]). One of ordinary skill in the art before the effective filing date of the claimed invention would have recognized that staged, decreasing pore size is a routine and predictable design in multi-layer filters that would yield the predictable results of efficient fractionation of blood components (MPEP 2143(I)(B)).
Regarding claim 18, Modified Menon teaches the method of claim 15.
Modified Menon fails to teach the third filter unit further includes additional filter unit according to a size of the protein to be separated. However primary reference, Menon, teaches the second layer to be an asymmetric membrane comprising larger pores on an upstream portion and smaller pores on a downstream portion, where the larger upstream pores act as a pre-filter while the smaller downstream pores serve as an absolute cut-off layer ([0085]). It would have been obvious for one of ordinary skill in the art before the effective filing date of the invention to have substituted the membrane used in the protein adsorption layer with an asymmetric membrane for the expectation of predictable results in order to reduce clogging in the subsequent layers ([0085]).
Regarding claim 19, Modified Menon teaches the method of claim 15, wherein the first filter unit is made of a material selected from a group of natural fibers (Any suitable size-exclusion filter can be used…an example…cellulose acetate size exclusion filter; Marble, [0038).
Regarding claim 20, Modified Menon teaches the method of claim 15, wherein the third filter unit is made of a material with low affinity and adsorption for nucleic acids (The protein adsorption layer 14 comprises a membrane conjugated to an 18-mer hydrocarbon chain (C18)…enables hydrophilic molecules, such as RNA/DNA, to flow through the layer 14 for subsequent preservation of protein-free nucleic acids; [0083]) or proteins.
Claims 8, 10, 16-17 are rejected under 35 U.S.C. 103 as being unpatentable over Menon et al. (US 20140273058 A1) with Pall Life Sciences (“Vivid™ Plasma Separation Membrane; 2009) incorporated by reference, in view of Marble (US 20130184446 A1), and in further view of Lee et al. (US 20150090674 A1), Miki et al. (US 20170258977 A1), Hillman et al. (US 5135719 A, 1990), and Fisher Scientific (“A clarifying guide to Membrane Filtration”; 2017).
Regarding claim 8, Modified Menon teaches the multi-layered filter according to claim 1.
Modified Menon is silent to teaching the first filter unit has a micron rating of 2-5 um, and a hole-up-volume of 20-100 uL.
Hillman teaches a micron rating of 2-5 µm (Particle size retention is preferably from about 1.0 to 3.0 microns; column 3, lines 66-67).
Hillman is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components and explores the use of filters capable of separating red blood cells from plasma starting with a whole blood sample (column 1, lines 59-61) as performed by the first filter described by Marble. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee and Miki to incorporate the teachings of Hillman by specifying the first filter to have a micron rating of 2-5 µm. Hillman states “particle size retention, composition of…thickness, and density should be taken into consideration in order to provide adequate filtration without hemolysis” (column 3, lines 59-62) as well as “to allow plasma to pass more rapidly through the filter than the red blood cells” (column 2, lines 2-3). In short, using the ranges of thickness with the corresponding particle size retention described by Hillman will prevent red blood cells from being destroyed and ultimately result in the partial separation of blood components from the whole blood sample. In the case where the claim ranges “overlap or lie inside ranges disclosed by the prior art” a prima facie case of obviousness exists (MPEP 2144.05(I)). It would have been obvious to try to optimize these result-effective variables due to the finite number of predictable potential solutions so as to achieve the maximum separation without cell damage (See MPEP 2144.05(I) and MPEP 2143(I)(E)).
Modified Menon is silent to teaching a hole-up-volume of 20-100 µL.
Fisher Scientific teaches a hole-up-volume of 20-100 µL (Improving Throughput table shows hold up volume column ranging from 10-80 µL with corresponding values for sample volume and, filter size and filtration area; page 18).
Fisher Scientific is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components and explores the use of filters capable of separating red blood cells from other components (page 4) as performed by the first filter described by Marble. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee, Miki, and Hillman to further incorporate the teachings of Fisher Scientific by specifying the first filter to have a hole-up-volume of 20-100 µL. As shown on page 18 by Fisher Scientific, the hold up volume is a function of the sample volume, filter size, and filtration area. The hold up volume can also depend on other parameters such as the material composition of the filtration, pressure, volume and the other parameters of the claim: weight, thickness, and micron rating. It would have been obvious before the effective filing date of the claimed invention to have tried to optimize the holdup volume range along with the other variables due to the finite number of predictable potential solutions so as to achieve better separation of whole blood cell components with a reasonable expectation of success (See MPEP 2143(I)(E) and MPEP 2144.05(I)).
Regarding claim 10, Modified Menon teaches the multi-layered filter according to claim 1, wherein the second filter unit has a micron rating of 1-5pm, and a hole-up-volume of 2-10 uL.
Modified Menon is silent to teaching the first filter unit has a micron rating of 2-5 pm, and a hole-up-volume of 20-100 uL.
Modified Menon does teach a hole-up-volume (void volume) of the asymmetric membrane is ~20 ± 1 μL/cm2 (see page 2, column 1, paragraph 2, lines 7-8 of the attached product data sheet for Pall Life sciences).
Hillman teaches a micron rating of 2-5 µm (Particle size retention is preferably from about 1.0 to 3.0 microns; column 3, lines 66-67).
Hillman is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components and explores the use of glass microfiber filters capable of separating red blood cells from plasma starting with a whole blood sample (column 1, lines 59-61) as performed by the first filter described by Marble. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee and Miki to incorporate the teachings of Hillman by substituting the second filter unit for a filter unit with a micron rating of 2-5 µm. Hillman states “particle size retention, composition…glass thickness, and density should be taken into consideration in order to provide adequate filtration without hemolysis” (column 3, lines 59-62) as well as “to allow plasma to pass more rapidly through the filter than the red blood cells” (column 2, lines 2-3). In short, using the micron rating range described by Hillman will prevent red blood cells from being destroyed and ultimately the partial separation of blood components from the whole blood sample which is a shared goal of reference Menon. It would have been obvious before the effective filing date of the claimed invention to optimize the well-known variable of micron rating to be 2-5 um so as to obtain the desired predictable result of efficient separation. (See MPEP 2144.05(I)), MPEP 2143(I)B)).
Modified Menon is silent to teaching a hole-up-volume of 2-10 uL.
Fisher Scientific teaches a hole-up-volume of 10-80 µL (Improving Throughput table shows hold up volume column ranging from 10-80 µL with corresponding values for sample volume and, filter size and filtration area; page 18).
Fisher Scientific is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components and explores the use of filters capable of separating red blood cells from other components (page 4) as performed by the second filter described by Menon. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee, Miki, and Hillman to incorporate the teachings of Fisher Scientific by specifying the second filter to have a hole-up-volume of 20-100 µL. As shown on page 18 by Fisher Scientific, the hold up volume is a function of the sample volume, filter size, and filtration area. The hold up volume can also depend on other parameters such as the material composition of the filtration, pressure, volume and the other parameters of the claim: weight, thickness, and micron rating. It would have been obvious before the effective filing date of the claimed invention to have optimized this variable of hole-up-volume to be 2-10 uL so as to obtain the desired result of efficient separation. (See MPEP 2144.05(I)).
Regarding claim 16, Modified Menon teaches the method of claim 15.
Modified Menon is silent to teaching the first filter unit has a micron rating of 2-5 um, and a hole-up-volume of 20-100 uL.
Hillman teaches a micron rating of 2-5 µm (Particle size retention is preferably from about 1.0 to 3.0 microns; column 3, lines 66-67).
Hillman is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components and explores the use of filters capable of separating red blood cells from plasma starting with a whole blood sample (column 1, lines 59-61) as performed by the first filter described by Marble. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee and Miki to incorporate the teachings of Hillman by specifying the first filter to have a micron rating of 2-5 µm. Hillman states “particle size retention, composition of…thickness, and density should be taken into consideration in order to provide adequate filtration without hemolysis” (column 3, lines 59-62) as well as “to allow plasma to pass more rapidly through the filter than the red blood cells” (column 2, lines 2-3). In short, using the ranges of thickness with the corresponding particle size retention described by Hillman will prevent red blood cells from being destroyed and ultimately result in the partial separation of blood components from the whole blood sample. In the case where the claim ranges “overlap or lie inside ranges disclosed by the prior art” a prima facie case of obviousness exists (MPEP 2144.05(I)). It would have been obvious to try to optimize these result-effective variables due to the finite number of predictable potential solutions so as to achieve the maximum separation without cell damage (See MPEP 2144.05(I) and MPEP 2143(I)(E)).
Modified Menon is silent to teaching a hole-up-volume of 20-100 µL.
Fisher Scientific teaches a hole-up-volume of 20-100 µL (Improving Throughput table shows hold up volume column ranging from 10-80 µL with corresponding values for sample volume and, filter size and filtration area; page 18).
Fisher Scientific is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components and explores the use of filters capable of separating red blood cells from other components (page 4) as performed by the first filter described by Marble. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee, Miki, and Hillman to further incorporate the teachings of Fisher Scientific by specifying the first filter to have a hole-up-volume of 20-100 µL. As shown on page 18 by Fisher Scientific, the hold up volume is a function of the sample volume, filter size, and filtration area. The hold up volume can also depend on other parameters such as the material composition of the filtration, pressure, volume and the other parameters of the claim: weight, thickness, and micron rating. It would have been obvious before the effective filing date of the claimed invention to have tried to optimize the holdup volume range along with the other variables due to the finite number of predictable potential solutions so as to achieve better separation of whole blood cell components with a reasonable expectation of success (See MPEP 2143(I)(E) and MPEP 2144.05(I)).
Regarding claim 17, Modified Menon teaches the method of claim 15.
Modified Menon is silent to teaching the first filter unit has a micron rating of 2-5 um, and a hole-up-volume of 20-100 uL.
Modified Menon does teach a hole-up-volume (void volume) of the asymmetric membrane is ~20 ± 1 μL/cm2 (see page 2, column 1, paragraph 2, lines 7-8 of the attached product data sheet for Pall Life sciences).
Hillman teaches a micron rating of 2-5 µm (Particle size retention is preferably from about 1.0 to 3.0 microns; column 3, lines 66-67).
Hillman is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components and explores the use of glass microfiber filters capable of separating red blood cells from plasma starting with a whole blood sample (column 1, lines 59-61) as performed by the first filter described by Marble. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee and Miki to incorporate the teachings of Hillman by substituting the second filter unit for a filter unit with a micron rating of 2-5 µm. Hillman states “particle size retention, composition…glass thickness, and density should be taken into consideration in order to provide adequate filtration without hemolysis” (column 3, lines 59-62) as well as “to allow plasma to pass more rapidly through the filter than the red blood cells” (column 2, lines 2-3). In short, using the micron rating range described by Hillman will prevent red blood cells from being destroyed and ultimately the partial separation of blood components from the whole blood sample which is a shared goal of reference Menon. It would have been obvious before the effective filing date of the claimed invention to optimize the well-known variable of micron rating to be 2-5 um so as to obtain the desired predictable result of efficient separation. (See MPEP 2144.05(I)), MPEP 2143(I)B)).
Modified Menon is silent to teaching a hole-up-volume of 2-10 uL.
Fisher Scientific teaches a hole-up-volume of 10-80 µL (Improving Throughput table shows hold up volume column ranging from 10-80 µL with corresponding values for sample volume and, filter size and filtration area; page 18).
Fisher Scientific is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components and explores the use of filters capable of separating red blood cells from other components (page 4) as performed by the second filter described by Menon. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee, Miki, and Hillman to incorporate the teachings of Fisher Scientific by specifying the second filter to have a hole-up-volume of 20-100 µL. As shown on page 18 by Fisher Scientific, the hold up volume is a function of the sample volume, filter size, and filtration area. The hold up volume can also depend on other parameters such as the material composition of the filtration, pressure, volume and the other parameters of the claim: weight, thickness, and micron rating. It would have been obvious before the effective filing date of the claimed invention to have optimized this variable of hole-up-volume to be 2-10 uL so as to obtain the desired result of efficient separation. (See MPEP 2144.05(I)).
Claim 9 is rejected under 35 U.S.C. 103 as being unpatentable over Menon et al. (US 20140273058 A1) with Pall Life Sciences (“Vivid™ Plasma Separation Membrane; 2009) incorporated by reference, in view of Marble (US 20130184446 A1), and in further view of Lee et al. (US 20150090674 A1), Miki et al. (US 20170258977 A1), Hillman et al. (US 5135719 A, 1990), Fisher Scientific (“A clarifying guide to Membrane Filtration”; 2017), and Silin et al. (US 20150053627 A1).
Regarding claim 9, Modified Menon teaches the multi-layered filter according to claim 8, wherein the pore size of the first filter unit is 10 um (the first size-exclusion filter has a pore size of about 10 µm; Marble, [0022]), the thickness range of the first filter unit includes 1000 um (“The filter member may have a thickness in a range from about 0.8 to about 1.2 mm,” wherein this equates to a range of 800-1200um; Lee, [0029]), the micron rating range of the first filter unit includes 3 um (Particle size retention is preferably from about 1.0 to 3.0 microns; Hillman, column 3, lines 66-67), and the hole-up-volume range of the first filter unit includes 50 uL (Improving Throughput table shows hold up volume column ranging from 10-80 µL with corresponding values for sample volume and, filter size and filtration area; Fisher Scientific, page 18).
Selecting the claimed thickness of 1000 um, micron rating of 3 pm, and hole-up-volume of 50 ul is optimization of a routine result-effective variable to yield the predictable outcome of a better separation of blood sample components (See MPEP 2144.05(I)). It would have been obvious before the effective filing date of the claimed invention to have chosen these filter characteristics so as to achieve such a benefit.
Modified Menon is silent to the teaching of the weight being 250 gram/m2.
Silin teaches the weight being 250 gram/m2 (the filter media may have a basis weight…equal to about 250 g/m.sup.2; ([0040]).
Silin is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components by using a multilayer filter ([0024][0085]). It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee, Miki, Hillman, and Fisher Scientific to further incorporate the teachings of Silin by specifying the first filter weight to be 250 gram/m2. Doing so will help to achieve “enhanced filtration properties” which is the ultimate goal of Menon for the separation of whole blood components. It would have been obvious before the effective filing date of the claimed invention to have optimized this variable of filter weight to be 250 g/m.sup.2 so as to obtain the desired result of efficient separation. (See MPEP 2144.05(I)).
Claim 11 is rejected under 35 U.S.C. 103 as being unpatentable over Menon et al. (US 20140273058 A1) with Pall Life Sciences (“Vivid™ Plasma Separation Membrane; 2009) incorporated by reference, in view of Marble (US 20130184446 A1), and in further view of Lee et al. (US 20150090674 A1), Miki et al. (US 20170258977 A1), Hillman et al. (US 5135719 A, 1990), Fisher Scientific (“A clarifying guide to Membrane Filtration”; 2017), Streiff (US 20190382699 A1), and Leland et al. (US 20180369473 A1).
Regarding claim 11, Modified Menon teaches the multi-layered filter according to claim 10.
Modified Menon fails to teach the pore size of the second filter unit is 0.1 µm, the thickness of the second filter unit is 300 um, the weight of the second filter unit is 200 gram/m2, the micron rating of the second filter unit is 2 um, and the hole-up-volume is 5 uL.
Streiff teaches the pore size of a filter unit is 0.1 µm (the size selective filter comprises pores with a diameter..of 0.1 μm; [0011]).
Streiff is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified Menon in view of Marble, Lee, Miki, Hillman, and Fisher Scientific to incorporate the teachings of Streiff by substituting the second filter unit for a filter unit with a pore size of 0.1 μm. Choosing an even smaller pore size (from 0.2 µm 0.1 µm) is still not smaller than the pore size of the third filter that is 0.01 µm. Therefore, adjusting the second filter pore size to 0.1 µm would serve the same purpose by yielding the predictable result of separating cell fragments, a function that is well known in the art (See MPEP 2143(I)(B)).
Modified Menon fails to teach the hole-up-volume is 5 µL.
Leland teaches the hole-up-volume is 5 µL (the dead volume will be about 5 μL; [0076]).
Leland is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have Modified Menon in view of Hillman and in further view of Silin to incorporate the teachings of Leland by substituting the first filter unit for a filter unit having a hole-up-volume of 5 µL. Leland already teaches “for optimal blood filtration, the pore size is recommended to be less than 2 microns” ([0058]) and further explains that “A low dead volume take-off maximizes the plasma recovery efficiency, which is necessary when the available blood volumes are limited” ([0037]). Adjusting the hole-up-volume to be 5 µL would serve the same purpose by yielding the predictable result of maximizing cell fractionation, a function that is well known in the art (See MPEP 2143(I)(B)).
Claims 21-22 are rejected under 35 U.S.C. 103 as being unpatentable over Menon et al. (US 20140273058 A1) with Pall Life Sciences (“Vivid™ Plasma Separation Membrane; 2009) incorporated by reference, in view of Marble (US 20130184446 A1), and in further view of Lee et al. (US 20150090674 A1), Miki et al. (US 20170258977 A1), and Valkirs et al. (US 20040219509 A1).
Regarding claim 21, Menon teaches a method of measuring blood P-selectin (whole blood; [0019]; See measurements in Figs. 7, 9-11) by using a disease diagnosis kit (all equipment in Figure 2) including a multi-layered filter that is made of a plurality of sequentially stacked filter units ((an integrated membrane assembly 30; [0087]; Fig. 2)( the layers 12, 14, 16 are stacked and pressed together as a single unit ; [0085]; Fig. 1; filters 32, 34, and 36 of Fig. 2), the method comprising:
separating and retaining blood platelets including P-selectin, ]) letting through a second passing-
through stream (“The porosity of the membrane was carefully selected in order to trap and remove cell/cellular debris while allowing nucleic acid species and proteins to flow through,” wherein cell is defined to include “platelets,” and since P-selectin is localized to platelets and platelet debris then P-selectin would also be separated; [0100][0069 through a first layer of the second filter unit (The sample collection layer 32 comprises an asymmetric membrane that has varying pore size (large to small) across the flow direction of the sample,” wherein the depth of larger pore size is the first layer; [0087]);
separating and retaining proteins in plasma including sP-selectin from the second passing-
through stream through a second layer of the second filter unit having a smaller pore size than the first layer of the second filter unit, letting through a third passing-through stream (“The porosity of the membrane was carefully selected in order to trap and remove cell/cellular debris while allowing nucleic acid species and proteins to flow through,” wherein cell is defined to include “platelets,” and since sP-selectin is localized to platelets and platelet debris then sP-selectin would also be separated; [0100][0069])(“The sample collection layer 32 comprises an asymmetric membrane that has varying pore size (large to small) across the flow direction of the sample,” wherein the depth of larger pore size is the first layer and the depth of the smaller pore size is the second layer);
separating and retaining small protein (a protein adsorption layer 84 designed to selectively remove proteins from the cell free flow through; [0100]) or nucleic acid components from the third passing through stream through a third filter unit letting through a remaining passing-through stream (“a second layer for protein adsorption,” or “a protein adsorption layer 34”; Abstract; [0087]; Fig. 2) and a fourth filter unit (a highly adsorbent RNA/DNA preservation layer 36 (alternatively described as the RNA/DNA adsorption layer); [0087]),
wherein the second, third, and fourth filter units are stacked in a particular sequential order in which the second filter unit is behind the first filter unit, the third filter unit is behind the second filter unit, and the fourth filter unit is behind the third filter unit, in a direction of the whole blood sample passing through the plurality of sequentially stacked filter units (sequence the filtering process to first filter cells and then proteins to avoid degradation of the nucleic acids; [0026]; See order of layers 32, 34, and 36 of Figs. 2-3)(the layers 12, 14, 16 are stacked and pressed together as a single unit ; [0085]; Fig. 1; filters 32, 34, and 36 of Fig. 2)(The top side 20 is exposed to whole blood while the bottom side 22 is exposed to the flow-through processed sample; [0075]; Fig. 1), detachably connected to each other (each holder 64 is capable of removably engaging at least one adjacent holder 64; [0091]; Figs. 2-3) with predetermined spaces separating therebetween (Paragraph [0091] explains that each layer 32, 34, and 36 is encased within its own holder 64 which has an upper and lower portion 66 and 68, wherein the lower portion of one holder snaps into the upper portion of the holder beneath it. Fig. 3 shows this housing to have a circular opening on the upper and lower portions revealing the filters (e.g. 36) in between which naturally creates a void space and is predetermined in that the reference chose to encase the filters)(See spacing being layers in Fig. 8), and disassembled after filtering (This pie-shaped design of the membrane 32, 34, 36 allows a user to easily separate sections of each of the membrane 32, 34, 36 for easy partial sample analysis; [0092]; Fig. 3), wherein the whole blood sample is separated into blood components by passing through each of the second, third, and fourth filter units in sequential order (See order of stacked filters 32, 34, and 36 of Fig. 2),
and wherein after the plurality of sequentially stacked filter units are disassembled after filtering the whole blood sample, the blood platelets including P-selectin retained at the first layer of the second filter unit are collected for a second further test or experiment pertaining to the blood platelets, the proteins in plasma including sP- selectin retained at the second layer of the second filter unit are collected for a third further test or experiment pertaining to the proteins in plasma, and the small protein or nucleic acid components retained at the third filter unit and the fourth filter unit are collected for a fourth further test or experiment pertaining to the small protein or nucleic acid components, or for a purpose of preservation or transportation via one or more sample-preservation methods (See the results of total protein concentration and RNA post-loading in Figs. 9-12)(This pie-shaped design of the membrane 32, 34, 36 allows a user to easily separate sections of each of the membrane 32, 34, 36 for easy partial sample analysis, leaving other sections of the membrane available for analysis at a later time or for concurrent analysis; [0092]), and the remaining passing-through stream include pure plasma components (plasma would naturally flow through the bottom of the fourth filter), and wherein the affinity-facilitating material of the fourth filter unit (36; [0083]; Fig. 1) is a silica material capable of absorbing and retaining nucleic acids (Silica membranes are an alternative membrane for nucleic acid capture due to their strong nucleic acid binding capacity; [0088])(DNA and RNA bind to silica matrices with high affinity; [0088]) and adsorbs nucleic acids and the one or more sample-reservation methods include drying and freezing (This extraction/preservation (thaw/freeze) cycle; [0097]).
Menon fails to teach:
measuring P-selectin, a first filter, having a pore size of 5-15 um, a thickness of 500-1500 um, and a weight of 100-400 gram/m2, separating blood cells from a whole blood sample, the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes,
and the blood cells retained at the first filter unit are collected for a first further test or experiment pertaining to the blood cells,
Valkirs teaches measuring P-selectin (detecting the level of P-selectin glycoprotein ligand-1 (PSGL-1) expressing T cells in a sample from the subject; Abstract)
Valkirs is considered to be analogous to the claimed invention because it is in the same field of endeavor for separating proteins in a blood sample. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modified the method taught by Modified Menon to incorporate the teachings of Valkirs by utilizing the multilayer filter to measure P-selectin in a blood sample. Reference Menon teaches that “information derived from the analysis of proteins and nucleic acids derived from blood and other fluids and tissues has provided early detection of and diagnosis capability for many medical conditions,” and the multi-layer filter disclosed in the reference provides a means for gathering this information. Valkirs recognizes that “test samples would be more readily analyzed following a fractionation or purification procedure, for example, separation of whole blood into serum or plasma components” ([0031]) when using P-selectin as biomarker to identify particular diseases ([0016]); however, utilizing the multi-layer filter taught by modified Menon would help to optimize this separation process. It would have been obvious to try utilizing the multilayer filter to measure P-selectin in a blood sample in order to meet the need for a useful disease diagnostic method. There are a finite number of identified predictable potential solutions for isolating P-selectin in a blood sample, and one of ordinary skill in the art would expect a reasonable expectation of success in doing so (MPEP 2143(I)(E)).
Modified Menon fails to teach:
a first filter, having a pore size of 5-15 um, a thickness of 500-1500 um, and a weight of 100-400 gram/m2 separating blood cells from a whole blood sample, the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes, and the blood cells retained at the first filter unit are collected for a first further test or experiment pertaining to the blood cells.
Marble teaches:
a first filter unit, having a pore size of 5-15 um, (the first size-exclusion filter has a pore size of about 10 µm; [0022]), separating blood cells (excludes passage therethrough of lymphocytes, monocytes, neutrophils, eosinophils, and basophils; [0015]) from a whole blood sample (the sample comprises whole blood; [0013])
Marble is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon to incorporate the teachings of Marble by adding a first filter above the second filter, having a pore size of 5-15 um, separating blood cells from a whole blood sample subjected to the same analysis as the other filter units, in order to reduce clogging in the subsequent layers and obtain information about the sample as a whole. Primary reference, Menon, teaches the second layer to be an asymmetric membrane comprising larger pores on an upstream portion and smaller pores on a downstream portion, where the larger upstream pores act as a pre-filter while the smaller downstream pores serve as an absolute cut-off layer ([0085]). Menon explains that this structure efficiently captures cells, including erythrocytes and platelets, while reducing clogging ([0085])(emphasis added). One of ordinary skill in the art would have been motivated to include a first, cell-separating filter above the asymmetric membrane in order to remove bulk whole blood cells early, protect finer downstream filters from fouling, and improve recovery of smaller analytes such as proteins and nucleic acids. The motivation is consistent with MPEP 2143(I)(A), as this is merely combining prior-art elements according to known methods to yield predictable results since both the first filter of Marble and the multilayer filter of Menon would each perform the same functions as they do separately.
Modified Menon fails to teach:
a first filter, having a thickness of 500-1500 um, and a weight of 100-400 gram/m2, the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes.
Lee teaches a filter having a thickness of 500-1500 um (“The filter member may have a thickness in a range from about 0.8 to about 1.2 mm,” wherein this equates to a range of 800-1200um; [0029]), and a weight of 100-400 gram/m2 (The filter member may have a weight per surface area (or a basis weight) in a range from about…70 to about 120 g/m.sup.2; [0029]).
Lee is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon in view of Marble to incorporate the teachings of Lee by adding a first filter, having a thickness of 500-1500 um and a weight of 100-400 gram/m2, in order to “to reduce the occurrence of hemolysis of corpuscles including erythrocytes” (Lee, [0028]). Primary reference, Menon expresses a desire to collect blood cells without lysis ([0101]). Secondary reference Marble teaches staged size-exclusion filters with the first filter dimensioned for whole-cell capture ([0015]). Lee teaches that blood filter members having a thickness and weight per surface area in a particular range provide high plasma recovery while preventing hemolysis ([0028][0029]). It would have been advantageous to select Lee’s disclosed thickness and weight for the first filter of Menon in view of Marble to ensure durability, capacity, and reduced cell lysis, as both Lee and Menon stress the importance of avoiding red blood cell rupture. The overlapping filter weight range between the teaching of Lee and the instant claim is a matter of optimizing a routine result-effective variable to yield the predictable outcome of a lower amount of lysed cells (See MPEP 2144.05(I)). It would have been obvious before the effective filing date of the claimed invention to have chosen these filter characteristics so as to achieve such a benefit.
Modified Menon fails to teach:
the first, third, and fourth filter units are stacked on an acrylic plate (emphasis added), and each of the filter units having different pore sizes.
Miki teaches filter layers stacked (see filter layers above 61 in Fig. 2) on an acrylic plate (second end plate 61; [0074]).
Miki is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon in view of Marble and Lee to incorporate the teachings of Miki by stacking the filter layers on an acrylic plate so as to provide additional rigid support using a lightweight material that is easily processed (Miki, [0074]). Substitution of the bottom portion 50 taught by Menon with the acrylic second endplate 61 of Miki would yield predictable results since they both serve the same purpose for providing a steady support for the device (See MPEP 2143(I)(B)).
Modified Menon fails to teach that each of the filter units has a different pore size.
Marble teaches that each filter unit has a different pore size (paragraphs [0006]-[0007] list pore size ranges that do not overlap for three different filters).
Marble is considered to be analogous to the claimed invention because it is in the same field of endeavor for a multi-layered filter for separating blood components. It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the multi-layered filter taught by Menon in view of Marble, Lee, Miki, and Valkirs to further incorporate the teachings of Marble by having filter units of different pore sizes “such that fluid passing from the proximal end to the distal end of the sample containment vessel passes through the first, second, and third filters” (Marble, [0006]). One of ordinary skill in the art before the effective filing date of the claimed invention would have recognized that staged, decreasing pore size is a routine and predictable design in multi-layer filters that would yield the predictable results of efficient fractionation of blood components (MPEP 2143(I)(B)).
Regarding claim 22, Modified Menon teaches the method of claim 21, wherein the whole blood sample is collected from stroke patients, and sP-selectin is found in the small proteins “Blood samples from patients with stroke,” wherein “P-selectin [is] also called granule membrane protein-140,” wherein paragraph [0075] of the instant publication US 20220168672 A1 states that “but when exposed to diseases such as stroke, it is converted to sP-selectin”; Valkirs, [0276];[0137]), separated, retained, and collected via the fourth further test or experiment (“The nucleic acid species remaining in the sample flow through were stabilized and preserved in the RNA/DNA adsorption layer 86,” which would thereby separate any extra blood components including small proteins from the nucleic acids; [0100])
Regarding claim 23, The multi-layered filter of claim 1, further comprising: a fourth filter unit , made of an affinity-facilitating material is a silica material capable of absorbing and retaining nucleic acids (Silica membranes are an alternative membrane for nucleic acid capture due to their strong nucleic acid binding capacity; [0088])(DNA and RNA bind to silica matrices with high affinity; [0088]) and adsorbs nucleic acids and the one or more sample-reservation methods include drying and freezing (This extraction/preservation (thaw/freeze) cycle; [0097]), separating and retaining a fourth set of blood components from the third passing-through stream, letting through a remaining passing-through stream, wherein the fourth set of blood components include small protein or nucleic acid components (the multi-layer filter taught by modified Menon is functionally capable of passing of amino acids, bases or various [metabolites] through the third filter. Menon states that the hydrophobic C18 filter of the protein adsorption layer allows nucleic acids to remain in the sample flow and are stabilized downstream in the fourth layer ([0079]), wherein the fourth filter unit is stacked behind the third filter unit in the direction of the whole blood sample passes through the multi-layered filter, and is detachably connected to the third filter unit with a predetermined space separating therebetween (Paragraph [0091] explains that each layer 32, 34, and 36 is encased within its own holder 64 which has an upper and lower portion 66 and 68, wherein the lower portion of one holder snaps into the upper portion of the holder beneath it. Fig. 3 shows this housing to have a circular opening on the upper and lower portions revealing the filters (e.g. 36)), and disassembled after filtering, wherein after the fourth filter unit is disassembled after filtering the whole blood sample, the fourth set of blood components retained at the fourth filter unit are collected for a fourth further test or experiment pertaining to the fourth set of blood components (See the results of total protein concentration and RNA post-loading in Figs. 9-12)(This pie-shaped design of the membrane 32, 34, 36 allows a user to easily separate sections of each of the membrane 32, 34, 36 for easy partial sample analysis, leaving other sections of the membrane available for analysis at a later time or for concurrent analysis; [0092]), or for a purpose of preservation or transportation via one or more sample-preservation methods, and the remaining passing-through stream include pure plasma components (plasma would naturally flow through the bottom of the fourth filter), and wherein the affinity-facilitating material of the fourth filter unit is a silica material capable of absorbing and retaining nucleic acids (Silica membranes are an alternative membrane for nucleic acid capture due to their strong nucleic acid binding capacity; [0088])(DNA and RNA bind to silica matrices with high affinity; [0088]), and the one or more sample-reservation methods include drying and freezing (This extraction/preservation (thaw/freeze) cycle; [0097]).
Regarding claim 24, The multi-layered filter of claim 15, further comprising: a fourth filter unit , made of an affinity-facilitating material is a silica material capable of absorbing and retaining nucleic acids (Silica membranes are an alternative membrane for nucleic acid capture due to their strong nucleic acid binding capacity; [0088])(DNA and RNA bind to silica matrices with high affinity; [0088]) and adsorbs nucleic acids and the one or more sample-reservation methods include drying and freezing (This extraction/preservation (thaw/freeze) cycle; [0097]), separating and retaining a fourth set of blood components from the third passing-through stream, letting through a remaining passing-through stream, wherein the fourth set of blood components include small protein or nucleic acid components (the multi-layer filter taught by modified Menon is functionally capable of passing of amino acids, bases or various [metabolites] through the third filter. Menon states that the hydrophobic C18 filter of the protein adsorption layer allows nucleic acids to remain in the sample flow and are stabilized downstream in the fourth layer ([0079]), wherein the fourth filter unit is stacked behind the third filter unit in the direction of the whole blood sample passes through the multi-layered filter, and is detachably connected to the third filter unit with a predetermined space separating therebetween (Paragraph [0091] explains that each layer 32, 34, and 36 is encased within its own holder 64 which has an upper and lower portion 66 and 68, wherein the lower portion of one holder snaps into the upper portion of the holder beneath it. Fig. 3 shows this housing to have a circular opening on the upper and lower portions revealing the filters (e.g. 36)), and disassembled after filtering, wherein after the fourth filter unit is disassembled after filtering the whole blood sample, the fourth set of blood components retained at the fourth filter unit are collected for a fourth further test or experiment pertaining to the fourth set of blood components (See the results of total protein concentration and RNA post-loading in Figs. 9-12)(This pie-shaped design of the membrane 32, 34, 36 allows a user to easily separate sections of each of the membrane 32, 34, 36 for easy partial sample analysis, leaving other sections of the membrane available for analysis at a later time or for concurrent analysis; [0092]), or for a purpose of preservation or transportation via one or more sample-preservation methods, and the remaining passing-through stream include pure plasma components (plasma would naturally flow through the bottom of the fourth filter), and wherein the affinity-facilitating material of the fourth filter unit is a silica material capable of absorbing and retaining nucleic acids (Silica membranes are an alternative membrane for nucleic acid capture due to their strong nucleic acid binding capacity; [0088])(DNA and RNA bind to silica matrices with high affinity; [0088]), and the one or more sample-reservation methods include drying and freezing (This extraction/preservation (thaw/freeze) cycle; [0097])
Conclusion
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/V.S./Examiner, Art Unit 1758
/MARIS R KESSEL/Supervisory Patent Examiner, Art Unit 1758
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