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 .
DETAILED ACTION
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
Election/Restrictions
Applicant’s election without traverse of Invention I, claims 1-69 and 158-164, in the reply filed on Apr. 17, 2026 is acknowledged.
Claims 1-69, 135 and 158-164 remain pending in the current application, claims 135 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention.
The requirement for the restriction of Inventions I and II is still deemed proper and is therefore made FINAL.
Claims 1-69 and 158-164 have been considered on the merits.
Status of the Claims
Claims 1-69, 135, and 158-164 are currently pending.
Claim 135 has been withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected Invention, there being no allowable generic or linking claim.
Claims 70-134 are cancelled.
Claims 1-69 and 158-164 have been considered on the merits.
Specification
The disclosure is objected to because of the following informalities: the use of trademarks.
The use of the terms Krytox™ GPL oil in 0072; Solvay® Galden® oil in 0072; Fluorinert™ FC-3283 and Fluorinert™ FC-40 in 0072, which are a trade names or a marks used in commerce, have been noted in this application. The terms should be accompanied by the generic terminology; furthermore the terms should be capitalized wherever they appear or, where appropriate, include a proper symbol indicating use in commerce such as ™, SM , or ® following the terms.
Although the use of trade names and marks used in commerce (i.e., trademarks, service marks, certification marks, and collective marks) are permissible in patent applications, the proprietary nature of the marks should be respected and every effort made to prevent their use in any manner which might adversely affect their validity as commercial marks.
Appropriate correction is required.
Claim Objections
The disclosure is objected to because of the following informalities:
Claim 158 is objected to because of the following informalities: the claim is missing a period at the end of it.
Appropriate correction is appreciated.
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 42-47 are 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.
In claim 42, the phrase “wherein a ratio between an average diameter of cells in the second ordered stream of cells and a width of the second microchannel is between 1 and 20”, renders the claim and its dependents indefinite, since it is unclear how the cell can be the same size or larger than the width of the microchannel. For the purposes of compact prosecution, the phrase in the claim will be interpreted to mean “wherein a ratio between a width of the second microchannel an average diameter of cells in the second ordered stream of cells
Since claims 43-47 depend from indefinite claim 42 and does not clarify the above points of confusion, claims 43-47 must also be rejected under 35 U.S.C. § 112, second paragraph.
Appropriate correction is required.
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
Claims 1-3, 6-7, 21-22, 36-38, 40-44, 46-47, 50-52, 57, 59, 61-62, 64, 68-69, 160 and 162 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Lagus et al. (RSC Advances, 2013) (ref. of record).
With respect to claims 1 and 69, Lagus teaches a method for encapsulating two cells in a single droplet (abstract). With respect to the first recited step of claims 1 and 69, Lagus teaches flowing a first aqueous suspension or phase containing a first ordered stream of cells, Cell Type A, in a first microchannel to a junction (Fig. 1, pg. 20513-20514 last para.). With respect to the second recited step of claims 1 and 69, Lagus teaches flowing a second aqueous suspension or phase containing a second ordered stream of cells, Cell Type B, in a second microchannel to a junction (Fig. 1, pg. 20513-20514 last para.). With respect to the third recited step of claims 1 and 69, Lagus teaches flowing an oil phase in a third microchannel to a junction (Fig. 1, pg. 20513-20514 last para.). With respect to the fourth recited step of claims 1 and 69, Lagus teaches at the junction generating the single droplet formed from the first aqueous phase, the second aqueous phase and the oil drop where the droplet contains a first stream and a second stream cell. Lagus teaches flowing together the first aqueous phase, the second aqueous phase, and the oil phase to generate the plurality of droplets (Fig. 1, pg. 20513-20514 last para.).
With respect to claim 2, Lagus teaches the fraction of droplets containing both a cell from the first stream and the second stream is higher than what would be predicted based on a Poisson distribution (abstract). With respect to claims 3 and 69, Lagus teaches the co-encapsulation of 64% in the droplets which is a nearly fivefold improvement to Poisson co-encapsulation and which is at least 20% of droplets in the plurality of droplets include a single cell from the first ordered stream of cells and at least one cell from the second ordered stream of cells (abstract).
With respect to claims 6 and 21, Lagus teaches the cells of the first and second ordered stream of cells are aligned along a central axis or edge of the first microchannel (Fig. 1 and 4 and pg. 20513-20415 bridging para.). With respect to claims 7 and 22, Lagus teaches that inertial effects in microfluidic channels allow for the formation of self-ordered trains of cells and teaches ordering channels placed upstream of the droplet-generating nozzle (pg. 20513 Col. 1 para. 2-4 and Fig. 1). Accordingly, Lagus teaches the first and second cells are aligned through inertial focusing while being flowed through the first microchannel and second microchannel, respectively.
With respect to claims 36-38 and 42-44, Lagus teaches channels have a width of 44 µm and the average cell diameter was 7.7±1.1 µm (pg. 20514 Col. 1 last para. and pg. 20517-20518 bridging para.). This gives ratio of a ratio between a width of the first microchannel and an average diameter of cells in the first ordered stream of cells of 5.7. With respect to claims 40 and 46, Lagus teaches the average cell diameter was 7.7±1.1 µm ( pg. 20517-20518 bridging para.). With respect to claims 41 and 47, Lagus teaches channels have a width of 44 µm (pg. 20514 Col. 1 last para.).
With respect to claims 50 and 51, Lagus teaches the method where generating the single droplet comprises: contacting the flowing first aqueous phase and the second aqueous phase with one another, where the contacting creates a single aqueous phase comprising the first ordered stream of cells and the second ordered stream of cells and where the contacting of the flowing first aqueous phase and the second aqueous phase to create the single aqueous phase occurs at a location at or prior to the junction (Fig. 1). With respect to claim 52, Lagus teaches generating the single droplet further comprises: contacting the flowing oil phase with the single aqueous phase to form a cone configuration within the junction, wherein the single droplet is generated at a tip of the cone configuration (Fig. 1).
With respect to claims 57 and 59, Lagus teaches the method where the aqueous flow rates in both channels 10 µL/min (pg. 20515 Col. 1 para. 2). With respect to claims 61 and 62, Lagus teaches the method the oil phase is flowed at a third rate of 45 µL/min (pg. 20515 Col. 1 para. 2). With respect to claims 64 and 68, Lagus teaches detecting the mating reaction between the algae cells in the droplets (the method further detecting an interaction between the cell from the first ordered stream of cells and the cell from the second ordered stream of cells and the detecting the interaction comprises detecting the interaction within the single droplet (pg. 20518 last para. to pg. 20519 para. 3 and Fig. 7).
With respect to claim 160, Lagus teaches the width of the aqueous channel is 75 µm, this is the junction of the channels (pg. 20514 Col. 1 last para. and Fig. 1). With respect to claim 162, Lagus teaches flowing the single droplet away from the junction through a nozzle region and flowing the single droplet through a post-nozzle region (Fig. 1 and Fig. 3a).
Therefore, the reference anticipates the claimed subject matter.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
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 1-3, 6-7, 15-17, 21-22, 30-32, 36-52, 57-62, 64, 68-69, and 160-164 are rejected under 35 U.S.C. 103 as being unpatentable over Lagus et al. (RSC Advances, 2013) (ref. of record) as evidenced by Amini et al. (US 2017/0128940 A1).
With respect to claims 1 and 69, Lagus teaches a method for encapsulating two cells in a single droplet (abstract). With respect to the first recited step of claims 1 and 69, Lagus teaches flowing a first aqueous suspension or phase containing a first ordered stream of cells, Cell Type A, in a first microchannel to a junction (Fig. 1, pg. 20513-20514 last para.). With respect to the second recited step of claims 1 and 69, Lagus teaches flowing a second aqueous suspension or phase containing a second ordered stream of cells, Cell Type B, in a second microchannel to a junction (Fig. 1, pg. 20513-20514 last para.). With respect to the third recited step of claims 1 and 69, Lagus teaches flowing an oil phase in a third microchannel to a junction (Fig. 1, pg. 20513-20514 last para.). With respect to the fourth recited step of claims 1 and 69, Lagus teaches at the junction generating the single droplet formed from the first aqueous phase, the second aqueous phase and the oil drop where the droplet contains a first stream and a second stream cell. Lagus teaches flowing together the first aqueous phase, the second aqueous phase, and the oil phase to generate the plurality of droplets (Fig. 1, pg. 20513-20514 last para.).
With respect to claim 2, Lagus teaches the fraction of droplets containing both a cell from the first stream and the second stream is higher than what would be predicted based on a Poisson distribution (abstract). With respect to claims 3 and 69, Lagus teaches the co-encapsulation of 64% in the droplets which is a nearly fivefold improvement to Poisson co-encapsulation and which is at least 20% of droplets in the plurality of droplets include a single cell from the first ordered stream of cells and at least one cell from the second ordered stream of cells (abstract).
With respect to claims 6 and 21, Lagus teaches the cells of the first and second ordered stream of cells are aligned along a central axis or edge of the first microchannel (Fig. 1 and 4 and pg. 20513-20415 bridging para.). With respect to claims 7 and 22, Lagus teaches that inertial effects in microfluidic channels allow for the formation of self-ordered trains of cells and teaches ordering channels placed upstream of the droplet-generating nozzle (pg. 20513 Col. 1 para. 2-4 and Fig. 1). Accordingly, Lagus teaches the first and second cells are aligned through inertial focusing while being flowed through the first microchannel and second microchannel, respectively.
With respect to claims 15-17 and 30-32, Lagus teaches the particle spacing was 24.9±5.3 µm in channel A and 23.7±4.3 µm in channel B (pg. 20516 Col. 2 para. 2). Lagus teaches the average cell diameter was 7.7±1.1 µm and the cell spacing was 25.7±8.8 µm in channel A and 26.8±12.2 µm in channel B (pg. 20517-20518 bridging para.). This gives the cell spacing about 3.3 times and 3.5 times an average cell diameter for channels A and B, respectively, and a standard deviation of a standard deviation of less than 10 µm inter-cell spacing between pairs of successive cells.
Lagus is silent with respect to whether these averages are representative for at least 80% of the cells in the first ordered and second order streams as recited in claims 15 and 30. Lagus does not teach an inter-cell spacing for at least 60% of cells in the first ordered and second order streams is between 1.5 times an average cell diameter and 3 times an average cell diameter as recited in claims 16 and 31. Similarly, Lagus is silent with the number of pairs of adjacent cells in the first ordered and second stream of cells which were measure to achieve the resulting average cell spacing and standard deviation and does not teach the spacing between pairs of successive cells is less than 10 µm when measured over 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 pairs of adjacent cells in the first ordered and second streams of cells as recited in claims 17 and 32.
Although Lagus does not teach the percentages of inter-cell spacing having the claimed ratios as recited in claims 15-16 and 30-31, the inter-cell spacing is between 1.5 times an average cell diameter and 3 times an average cell diameter as recited in claims 16 and 30, and the standard deviation of the cell spacing is less than to 10 µm when measured over 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 pairs of adjacent cells in the first ordered and second ordered streams of cells, one of ordinary skill in the art would recognize that the percentage of inter-cell spacings having a particular ratio and a certain standard deviation and inter-cell spacing are result effective variables and that these parameters would be matter of routine optimization as evidenced by Lagus. Lagus reports that at sufficiently high mean flow velocity and cell diameter, cells migrate to lateral equilibrium positions and form self-ordered trains of equal longitudinal spacing (pg. 20513 Col. 1 para. 1). Lagus reports train spacing (inter-cell spacing) can be increased or decreased by using lower or higher particle concentration, respectively (pg. 20516 Col. 1 last para. to Col. 2 para. 2). Accordingly, one of ordinary skill in the art would adjust the flow velocity, diameter of channels (depending on the cell diameter) and concentration of the cell solution to control for the inter-cell spacing within the channel.
With respect to claims 36-38 and as claims 42-44 are being interpreted (see rejections under 35 U.S.C §112(b)), Lagus teaches channels have a width of 44 µm and the average cell diameter was 7.7±1.1 µm (pg. 20514 Col. 1 last para. and pg. 20517-20518 bridging para.). This gives ratio of a ratio between a width of the first microchannel and an average diameter of cells in the first and second ordered streams of cells of 5.7.
Lagus does not teach a ratio between an average width of the first or second microchannel and an average diameter of cells in the first or second ordered stream of cells is between 2.5 and 5.0 as recited in claim 39 and as claim 45 is being interpreted (see rejections under 35 U.S.C §112(b)). Although Lagus does not teach the claimed range of the ratio between the width of microchannel and the diameter of the cell, one of ordinary skill in the art would recognize that this ratio is a result effective variable and would be matter of routine optimization as evidenced by Amini. Amini reports that in methods of ordering particles such as cells in microfluidic devices that the closer the size of the channel in is to the bead size or particle size, the faster and more efficient the separating, focusing and ordering (abstract and 0042).
With respect to claims 40 and 46, Lagus teaches the average cell diameter was 7.7±1.1 µm (pg. 20517-20518 bridging para.). With respect to claims 41 and 47, Lagus teaches channels have a width of 44 µm (pg. 20514 Col. 1 last para.).
Although Lagus does not explicitly teach the maximum concentration of the first order stream of cells or the second ordered stream cells is defined by the formal recited in claims 48 and 49, these equations would be readily recognized by one of ordinary skill in the art to be adapted depending on the rate of the fluid movement through the channels to be determine the maximum concentration of cells.
With respect to claims 50 and 51, Lagus teaches the method where generating the single droplet comprises: contacting the flowing first aqueous phase and the second aqueous phase with one another, where the contacting creates a single aqueous phase comprising the first ordered stream of cells and the second ordered stream of cells and where the contacting of the flowing first aqueous phase and the second aqueous phase to create the single aqueous phase occurs at a location at or prior to the junction (Fig. 1). With respect to claim 52, Lagus teaches generating the single droplet further comprises: contacting the flowing oil phase with the single aqueous phase to form a cone configuration within the junction, wherein the single droplet is generated at a tip of the cone configuration (Fig. 1).
With respect to claims 57 and 59, Lagus teaches the method where the aqueous flow rates in both channels 10 µL/min (pg. 20515 Col. 1 para. 2). With respect to claims 61 and 62, Lagus teaches the method the oil phase is flowed at a third rate of 45 µL/min (pg. 20515 Col. 1 para. 2).
Lagus does not teach method where the first aqueous phase or where the second aqueous phase is flowed at a first rate of about 45 µL/min as recited in claims 58 and 60. Although Lagus does not teach the claimed flow rate for the first and second aqueous phases, one of ordinary skill in the art would recognize that flow rate is a result effective variable and would be matter of routine optimization as evidenced by Amini. Amini reports that inter-bead spacing is determined by fluid and flow parameters, geometric parameters such as bead diameter, channel width and height (0054). Amini teaches “different channel geometries and flow rates can be used for the streams of cell flow and cell fluid to ensure desired focusing and ordering in each stream prior to droplet generation” (0072).
With respect to claims 64 and 68, Lagus teaches detecting the mating reaction between the algae cells in the droplets (the method further detecting an interaction between the cell from the first ordered stream of cells and the cell from the second ordered stream of cells and the detecting the interaction comprises detecting the interaction within the single droplet (pg. 20518 last para. to pg. 20519 para. 3 and Fig. 7).
With respect to claim 160, Lagus teaches the width of the aqueous channel is 75 µm, this is the junction of the channels (pg. 20514 Col. 1 last para. and Fig. 1).
Lagus is silent with respect to the width of the third channel and does not teach that it is between 5 to 500 µm as recited in claim 161. However, one of ordinary skill in the art would recognize width of the third channel is a result effective variable and would be matter of routine optimization as evidenced by Amini. Amini reports that oil inlet is configured for droplet generation at the junction and the flow is matched with the frequency of the stream aqueous cell fluid (0075). Additionally, Amini teaches “different channel geometries and flow rates can be used for the streams of cell flow and cell fluid to ensure desired focusing and ordering in each stream prior to droplet generation” (0072).
With respect to claim 162, Lagus teaches flowing the single droplet away from the junction through a nozzle region and flowing the single droplet through a post-nozzle region (Fig. 1 and Fig. 3a).
With respect to claim 163, Lagus teaches the width of the nozzle is 44 µm (pg. 20514 Col. 1 last para. and Fig. 1).
However, Lagus is silent with respect to the length of the nozzle and does not teach that it is between 20 to 500 µm as recited in claim 163. Similarly Lagus does not teach the method where the width of the post-nozzle region is between 50 pm to 1000 µm as recited in claim 164. However, one of ordinary skill in the art would recognize width of the length of the nozzle and the width of the post-nozzle region are result effective variables and would be matter of routine optimization as evidenced by Amini. Amini reports that the output region or post-nozzle region shape can be altered to have a straight region depending on the function such as having a region for analysis (0046). Additionally, one of ordinary skill in the art would readily recognize that the width of the nozzle would be optimized for the size of the droplets being produced.
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective time of filing of the invention, especially in the absence of evidence to the contrary.
Claims 4-5, 53-56, and 65-67 are rejected under 35 U.S.C. 103(a) as being unpatentable over Lagus as evidenced by Amini (as applied to claims 1-3, 6-7, 15-17, 21-22, 30-32, 36-52, 57-62, 64, 68-69, and 160-164 above), and further in view of Weitz et al. (US 2016/0201129 A1) (ref. of record).
The teachings of Lagus can be found in the previous rejection above.
Lagus is silent with respect to the rate of generation of the droplets and does not teach the method generates single droplets at a rate of at least 5,000 droplets per second as recited in claim 4, or at a rate of at least 8,000 droplets per second as recited in claim 5.
However, Weitz teaches a similar method of encapsulating two or more cells in a single droplets using microfluidic devices (abstract and 0027-0028). With respect to claim 4, Weitz teaches droplets can be generated at relatively high rates and are generated at rate of at least 5000 droplets per second (0047). With respect to claim 5, Weitz teaches droplets are generated at rate of at least 10,000 droplets per second (0047).
Accordingly, at the effective time of filing of the claimed invention, one of ordinary skill in the art would have been motivated to modify the method of Lagus so that the rate of droplet formation is at least 5,000 or 8,000 droplets per second for the benefit of producing droplets containing cells at a relatively high rate as taught by Weitz. It would have been obvious to one of ordinary skill to modify the method of Lagus so that the rate of droplet formation a rate known in the art for forming droplets containing cells such as at least 5,000 or 8,000 droplets per second as taught by Weitz. For reason that the rates of at least 5,000 or 8,000 droplets per second was known to successfully to form droplets containing cells in the art, one of ordinary skill in the art would have a reasonable expectation of success.
Lagus does not teach the method where the cells from the first ordered stream of cells and the cells from the second ordered stream of cells are different cells as recited in claim 53. Lagus does not teach the method where the cell from the first ordered stream of cells is a T-cell as recited in claim 54. Lagus does not teach the method where the cell from the second ordered stream of cells is an antigen presenting cell (APC) as recited in claim 55. Lagus does not teach the method where the single droplet further comprises at least a second cell from the second ordered stream of cells as recited in claim 56.
However, Weitz teaches a similar method of encapsulating two or more cells in a single droplets using microfluidic devices (abstract and 0027-0028). With respect to claim 53, Weitz teaches the droplets containing two different cells (0030 and 0096). With respect to claim 54, Weitz teaches one of the cells is a T-cell (0030 and 0096). With respect to claim 55, Weitz teaches one of the cells is a B-cell or macrophages (antigen presenting cells (APC)) (0031 and 0096). With respect to claim 56, Weitz teaches the droplets can contain two or more cells where some of the cells may be substantially the same and/or there may be three, four, or more types of cells contained within the droplet (0029)
At the time of the claimed invention, one of ordinary skill in the art would have been motivated to modify the teachings of Lagus in such a way that the first and second streams contain different cells, that the first ordered stream contains T cells, the second order stream contains antigen presenting cells, and where there is at least of one of the cell types for the purpose being able form a droplet containing at least 2 different cells types, to form a droplet where one of the cells is a T cell and to form a droplet where one of the cells is an APC. Furthermore, it would have been obvious to one skilled in the art to have further modified the method of Lagus to form a droplet containing at least two cells such that the first and second streams contain different cells, that the first ordered stream contains T cells and the second order stream contains antigen presenting cells for the purpose being able form a droplet containing 2 different cells types, contain a T cell, contain an APC, or where there is at least of one of the cell types, since methods forming droplets with at least two cells were known to have two different cells types, contain T cells and contain APCs as taught by Weitz. Such a modification merely involves the substitution of one known type of configuration for another for the forming of droplets containing cells and for that reason there would be a reasonable expectation of success.
Lagus does not teach the method where the first aqueous phase or the second aqueous phase further comprise reagents for detecting the interaction as recited in claim 65. Similarly, Lagus does not teach the method where the reagents comprise any of fluorescent markers, beads, or nucleic acid barcodes as recited in claim 66. Lagus does not teach the method where detecting the interaction comprises detecting a biomarker analyte indicative of the interaction as recited in claim 67.
However, Weitz teaches similar method of encapsulating two cells in a single droplet and determining the effect immune cells have on target cells in the droplets (the method further detecting an interaction between the cell from the first ordered stream of cells and the cell from the second ordered stream of cells and the detecting the interaction comprises detecting the interaction within the single droplet) (abstract, Fig. 2, Fig. 5A and 0016). With respect to claims 65 and 66, Weitz teaches the method where the droplets comprise reagents that affect the interaction between the cells such as fluorescent species, dyes and nanoparticles (beads) (0049 and 105-0107) for detecting the interaction. With respect to claims 67, Weitz teaches the method where detecting the interaction comprises detecting an antibody (a biomarker analyte indicative of the interaction) (0108-0110).
Accordingly, at the effective time of filing of the claimed invention, one of ordinary skill in the art would have been motivated to modify the method of Lagus so that the first aqueous phase or the second aqueous phase further comprise reagents such as fluorescent markers, beads, or nucleic acid barcodes for detecting the interaction and detecting a biomarker analyte indicative of the interaction for the benefit of analyzing the reaction between the cells in the droplet as taught by Weitz. It would have been obvious to one of ordinary skill to modify the method of Lagus so that to include having either the first aqueous phase or the second aqueous phase further contain reagents such as fluorescent markers, beads, or nucleic acid barcodes for detecting the interaction between the cells and detecting a biomarker analyte indicative of the interaction, since similar methods of encapsulating at least two cells in a droplet and detecting an interaction between the cells include adding reagents to the aqueous phases and detecting an analyte to determine the interaction between the cells as taught by Weitz. Additionally, for this reason, one of ordinary skill in the art would have a reasonable expectation of success.
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective time of filing of the invention, especially in the absence of evidence to the contrary.
Claims 8-14 and 23-29 are rejected under 35 U.S.C. 103(a) as being unpatentable over Lagus as evidenced by Amini (as applied to claims 1-3, 6-7, 15-17, 21-22, 30-32, 36-52, 57-62, 64, 68-69, and 160-164 above), and further in view of Martel et al. (Annual review of biomedical engineering, 2014) (ref. of record).
The teachings of Lagus can be found in the previous rejection above.
Lagus does not teach the method where the inertial focusing is generated by flowing the first aqueous phase or a second aqueous phase through a curved region of the first microchannel or second microchannel, respectively, as recited in claims 8 and 23. Likewise, Lagus does not teach the method where wherein the curved region is between 150-300 mm in length as recited in claims 9 and 24, wherein the curved region is between 50-150 mm in length as recited in claims 10 and 25, or wherein the curved region is about 100 mm in length as recited in claims 11 and 26.
However, Martel teaches a method of ordering particles in microfluidic devices and where there is an addition of curvature to a microchannel pathway to generate secondary flows (pg. 372 para. 1-2 and pg. 375 para. 4). Martel further teaches that Dean flow allows for separation based on size due to the dependence of the drag force on particle diameter and the balance of this drag force with the shear gradient lift (pg. 380). In further support, Lagus teaches that curved channels, which induced secondary Dean flows that pushed cells to one side of the ordering channel (pg. 20513 Col. 1 para. 3).
Accordingly, at the effect of time of filing of the claimed invention one of ordinary skill in the art would have been motivated to modify the method of Lagus to include inertial focusing by flowing the aqueous phases through a curved region for the benefit of ordering the cells in microfluidic channels as taught by Martel. It would have been obvious to one of ordinary skill in the art to include curves in the microfluid device used in the method of Lagus, since curved microfluidic channels were known for providing inertial focusing and order of particle flowing the channel as taught by Martel. Furthermore, one of ordinary skill in the art would have had a reasonable expectation of succuss in making such a modification to the method of Lagus, since inertial focusing by flowing an aqueous phase containing particles was known to use curved channels as taught by Martel.
Martel does not teach that the curved region is between 150-300 mm in length as recited in claims 9 and 24, wherein the curved region is between 50-150 mm in length as recited in claims 10 and 25, or wherein the curved region is about 100 mm in length as recited in claims 11 and 26. However, one of ordinary skill in the art would recognize that the length of the curved region is a result effective variable and that the length of the curved region would be matter of routine optimization as evidenced by Martel. Martel teaches channel length needs to be adjusted to control or achieve equilibrium positions in inertial focusing (pg. 382-383 bridging para.).
Lagus does not teach the method where the curved region comprises at least one undulating portion comprising at least a 45 degree change in a flow vector across a length of the undulating portion as recited in claims 12 and 27, where the curved region comprises at least one undulating portion comprising at least a 60 degree change, at least a 90 degree change, at least a 120 degree change, at least a 150 degree change, or at least a 180 degree change in a flow vector across a length of the undulating portion as recited in claims 13 and 28, where the curved region comprises between 60-120 undulating portions as recited in claims 14 and 29.
However, Martel teaches a method of ordering particles in microfluidic devices and where there is an addition of curvature to a microchannel pathway to generate secondary flows (pg. 372 para. 1-2 and pg. 375 para. 4). With respect to claims 12 and 27, Martel teaches a region of the microchannel pathway containing at least one asymmetrically curved or undulating portion with at least a 45 degree change in a flow vector across a length of the asymmetrically curve or undulating portion (Fig. 1A). With respect to claims 13 and 28, Martel teaches the asymmetrically curve or undulating portion where there is nearly a 180 degree change and a 90 degree change in the flow vector across the length of the asymmetrically curve or undulating portion (with at least one undulating portion comprising at least a 60 degree change, at least a 90 degree change, at least a 120 degree change, at least a 150 degree change, or at least a 180 degree change in a flow vector across a length of the undulating portion) (Fig. 1A).
Accordingly, at the effect of time of filing of the claimed invention one of ordinary skill in the art would have been motivated to modify the method of Lagus to include inertial focusing by flowing the aqueous phases through a curved region containing undulating portions for the benefit of inertial focusing or ordering of the cells in microfluidic channels as taught by Martel. It would have been obvious to one of ordinary skill in the art to include asymmetrically curves or undulating portions in the microfluid channels used in the method of Lagus, since asymmetrically curves or undulating portions in microfluidic channels were known for providing inertial focusing and order of particle flowing the channel as taught by Martel. Furthermore, one of ordinary skill in the art would have had a reasonable expectation of succuss in making such a modification to the method of Lagus, since inertial focusing by flowing an aqueous phase containing particles was known to use asymmetrically curves or undulating portions as taught by Martel.
Martel is silent with respect to the number of undulating portions and does not teach the method where the curved region comprises between 60-120 undulating portions as recited in claims 14 and 29. However, one of ordinary skill in the art would recognize that the number of undulating portions is a result effective variable and that the number of undulating portions would be matter of routine optimization as evidenced by Martel. Martel teaches combining asymmetrical curves as well as spiral systems are all cases in which the forces change as the geometry changes along the length of the channel (pg. 382 para. 2).
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective time of filing of the invention, especially in the absence of evidence to the contrary.
Claims 18-20 and 33-35 are rejected under 35 U.S.C. 103(a) as being unpatentable over Lagus as evidenced by Amini (as applied to claims 1-3, 6-7, 15-17, 21-22, 30-32, 36-52, 57-62, 64, 68-69, and 160-164 above), and further in view of Zhang et al. (Lab on a Chip, 2016).
The teachings of Lagus can be found in the previous rejection above.
Lagus does not teach the method where the inter-cell spacing between the pairs of cells in the first ordered stream of cells or the second stream of cells is modulated by passing the pairs of cells through a set of pillars as recited in claims 18 and 33, where the set of pillars is positioned at an entrance of the first microchannel or the second microchannel as recited in claims 19 and 34, or where the set of pillars at the entrance of the first microchannel or second channel comprise 5 to 40 µm gaps between pillars as recited in claims 20 and 35.
However, Zhang teaches methods of inertial microfluidics to focus, concentrate and separate particles and for cellular sample processing (abstract). Zhang reviews microchannel structures and the mechanisms and under lying physics in inertial microfluidic systems (abstract). With respect to claims 18 and 19, Zhang teaches a method of ordering particles in microfluidic device where the channel contains pillars (Fig. 9d). Zhang teaches the introduction of obstacles in straight channels induces convective secondary flow (pg. 25 Col. 1 last para.). Zhang teaches that micropillars within a micro-channel will induce irreversible twisted flows at a finite inertial flow (pg. 27 Col. 2 para. 2). Zhang teaches that “the lateral position of the pillar can be used to tune the position and shape of the net recirculating flows across the channel” (pg. 27 Col. 2 para. 2). Zhang teaches that sequenced micropillars can be used to modify the inertial migration progress and reports that sequential micropillars have been used aid the single-stream focusing of microparticles (pg. 27 Col. 2 para. 3). Zhang teaches a device where blood cells are flowed through an array of circular pillars with specific gaps so that deterministic lateral displacement (DLD) size-based hydrodynamic force filters smaller red blood cells from white blood cells and circulating tumor cells (CTCs) with much larger size (pg. 30 Col. 1 para. 2). With respect to claims 19 and 34, Zhang teaches this method of flowing blood cells where the pillars are at the entrance of the microchannel to cause the separation of different cell types (Fig. 10b).
Accordingly, at the effect of time of filing of the claimed invention one of ordinary skill in the art would have been motivated to modify the method of Lagus to include a set of pillars at the entrance of the first and second microchannels for the benefit of separating different cells if the fluid contains multiple cell types and to further order the cells in the streams as taught by Zhang. It would have been obvious to one of ordinary skill in the art to include a set of pillars at the entrance of the first and second microchannels in the microfluid channels used in the method of Lagus, since pillars were known for providing separation of cells and further ordering of cells and particles flowing through channels as taught by Zhang. Furthermore, one of ordinary skill in the art would have had a reasonable expectation of succuss in making such a modification to the method of Lagus, since inertial focusing and separation by flowing an aqueous phase containing cells or particles was known to use pillars at the entrance of the microchannel as taught by Zhang.
Although, Zhang is silent with respect to the set of pillars at the entrance of the first microchannel or second channel comprise 5 to 40 µm gaps between pillars as recited in claims 20 and 35, one of ordinary skill in the art would recognize that spacing between the pillars within the channel is a result effective variable and would be matter of routine optimization as evidenced by Zhang. Zhang teaches a device where blood cells are flowed through an array of circular pillars with specific gaps so that deterministic lateral displacement (DLD) size-based hydrodynamic force filters smaller red blood cells from white blood cells and circulating tumor cells (CTCs) with much larger size (pg. 30 Col. 1 para. 2). Additionally, Zhang teaches that “the lateral position of the pillar can be used to tune the position and shape of the net recirculating flows across the channel” (pg. 27 Col. 2 para. 2).
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective time of filing of the invention, especially in the absence of evidence to the contrary.
Claims 63, 158 and 159 are rejected under 35 U.S.C. 103(a) as being unpatentable over Lagus as evidenced by Amini (as applied to claims 1-3, 6-7, 15-17, 21-22, 30-32, 36-52, 57-62, 64, 68-69, and 160-164 above), and further in view of Amini et al. (US 2017/0128940 A1).
The teachings of Lagus can be found in the previous rejection above.
Lagus does not teach the method where the second aqueous phase is flowed at a second rate that is faster than a first rate of the first aqueous phase, such that the single droplet comprises only a single cell from the first ordered stream of cells and two or more cells from the second ordered stream of cells as recited in claim 63.
However, Amini teaches a similar method of encapsulating cells in a single droplet using microfluidic devices (abstract). With respect to claim 63, Amini teaches that the flow rates for the first and second particle streams can be the same or different (0166). Amini further teaches the average number of cells and beads in each droplet is related to the flow rate of the different channels (0009). Amini teaches that “different channel geometries and flow rates can be used for the streams of cell flow and cell fluid to ensure desired focusing and ordering in each stream prior to droplet generation” (0072).
Accordingly, at the effect of time of filing of the claimed invention one of ordinary skill in the art would have been motivated to modify the method of Lagus so that the flow rate of the second ordered stream is faster than the first ordered stream for the benefit of producing droplets containing only a single cell from the first ordered stream of cells and two or more cells from the second ordered stream of cells as taught by Amini. It would have been obvious to one of ordinary skill in the art to adjust the flow rate of the channels to control the proportion of the cells from in each channel in the method of Lagus so that the second aqueous phase is flowed at a second rate that is faster than a first rate of the first aqueous phase, such that the single droplet comprises only a single cell from the first ordered stream of cells and two or more cells from the second ordered stream of cells, since this was well known in art to control the flow rate of the channels to control the number of beads or cells in the droplets as taught by Amini. Furthermore, one of ordinary skill in the art would have had a reasonable expectation of succuss in making such a modification to the method of Lagus, since the relationship of flow rate proportion of cells from each channel was known and having channels with different flow rates was known as taught by Amini.
Lagus does not teach the method where the first microchannel tapers or the second microchannel tapers down from a first width between 40 µm to 100 µm to a second width between 10 µm to 40 µm as the first microchannel approaches the junction as recited in claims 158 and 159, respectively.
However, Amini teaches the method where the microchannel dimensions decrease over the length to facilitate the filtering of the sample or to create fluidic resistance (0063 and 0072).
Accordingly, at the effect of time of filing of the claimed invention one of ordinary skill in the art would have been motivated to modify the method of Lagus so that microchannels taper down as they approach the junction for the benefit of filtering of the sample or to create fluidic resistance in the system as taught by Amini. It would have been obvious to one of ordinary skill in the art to modify the channels to taper as they approach the junction in the method of Lagus, since similar microfluidic systems for forming droplets of cells were known to have channels that taper as they approach as they approach the junction as taught by Amini. Furthermore, one of ordinary skill in the art would have had a reasonable expectation of succuss in making such a modification to the method of Amini, since such dimensional changes were known to be used as taught by Amini.
Although, Amini is silent with respect to the final width of the channels at the junction and does not teach the method where the first microchannel tapers or the second microchannel tapers down from a first width between 40 µm to 100 µm to a second width between 10 µm to 40 µm as the first microchannel approaches the junction as recited in claims 158 and 159, one of ordinary skill in the art would recognize that final width of the channels is a result effective variable and would be matter of routine optimization as evidenced by Amini. Amini teaches that the channel height to width aspect ratio may be select to optimize particle ordering (0071). Amini reports that inter-bead spacing is determined by fluid and flow parameters, geometric parameters such as bead diameter, channel width and height (0054). Additionally, Amini teaches the channels can have different geometries including varying diameter with an expansion/contraction region after a cell focusing region to enable the adjustment of the spacing between cells inside the channel (0072).
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective time of filing of the invention, especially in the absence of evidence to the contrary.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the claims at issue are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); and In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on a nonstatutory double patenting ground provided the reference application or patent either is shown to be commonly owned with this application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The USPTO internet Web site contains terminal disclaimer forms which may be used. Please visit http://www.uspto.gov/forms/. The filing date of the application will determine what form should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to http://www.uspto.gov/patents/process/file/efs/guidance/eTD-info-I.jsp
Claims 1-17, 21-32, 36-55, 57-62, 69 and 158-164 are rejected on the ground of nonstatutory obviousness-type double patenting as being unpatentable over claims 1-6, 8, 10, 11, 15, 17 and 18 of U.S. Patent No. 12,391,914. Although the conflicting claims are not identical, they are not patentably distinct from each other because the instant claims encompass those of the issue patent. In addition, both claim methods for encapsulating two cells in a single droplet.
The limitations of instant claims 1 and 69, are recited by claim 1 of Pat. No. 12,391,914. Specifically, claim 1 of Pat. No. 12,391,914 recites flowing a first aqueous suspension or phase containing a first ordered stream of cells in a first microchannel to a junction; flowing a second aqueous suspension or phase containing a second ordered stream of cells in a second microchannel to a junction; flowing an oil phase in a third microchannel to a junction; and at the junction generating the single droplet formed from the first aqueous phase, the second aqueous phase and the oil drop where the droplet contains a first stream and a second stream cell.
The limitations of instant claim 2, are recited by claim 1 of Pat. No. 12,391,914 which recites the fraction of droplets containing both a cell from the first stream and the second stream is higher than what would be predicted based on a Poisson distribution (abstract). Although the claims of Pat. No. 12,391,914 do not recited the method wherein the fraction exceeds the predicted fraction by a factor ranging from 2-3 as recited in instant claim 3, one of ordinary skill in the art would recognize that this fraction of cells produced containing one of each cell type is a result effective variable which can be adjusted by controlling different factor in the system such as the flow rates of the fluids in the different microchannels.
The limitations of instant claim 4, are recited by claim 2 of Pat. No. 12,391,914 where the method generates single droplets at a rate of at least 5,000 droplets per second. Although the claims of Pat. No. 12,391,914 do not recited the method generates single droplets at a rate of at least 8,000 droplets per second as recited in instant claim 5, one of ordinary skill in the art would recognize the rate of droplet formation would be a matter of routine optimization depending on different factors such as the rate at which the individual fluids flow, the composition of the fluids and the desired production rate.
The limitations of instant claim 7, is recited by claim 10 of Pat. No. 12,391,914 where the cells of the first ordered stream are aligned through inertial focusing while flowing through the channels. Although the claims of Pat. No. 12,391,914 do not recited the method where the cells of the first and second ordered stream of cells are aligned along a central axis or edge of the first and second microchannel as recited in instant claims 6 and 21, or where the cells of the second ordered stream are aligned through inertial focusing while flowing through the channels as recited in instant claim 22, by practicing the method of claim 1 of Pat. No. 12,391,914 where the cells are flowed through the undulating regions this would inherently occur.
The limitations of instant claims 8, 12-14, 23, and 27-29, are recited by claims 1 3, and 4 of Pat. No. 12,391,914 where the first and second microchannels comprises between 60-120 undulating portions where the portions comprising at least a 90 degree change in a flow vector across a length of the undulating portion. These undulating portions are curves. Although the claims of Pat. No. 12,391,914 do not recited the method where the wherein the curved region is between 150-300 mm in length as recited in instant claims 9 and 24, wherein the curved region is between 50-150 mm in length as recited in instant claims 10 and 25, or wherein the curved region is about 100 mm in length as recited in instant claims 11 and 26, one of ordinary skill in the art would recognize the length of the curved region would be a matter of routine optimization depending on different factors such as the cell size and to control or achieve equilibrium positions in inertial focusing.
The limitations of instant claims 15, 16, 30 and 31, are recited by claim 18 of Pat. No. 12,391,914 where an inter-cell spacing for at least 80% of cells in the first and second ordered streams is between 1.5 times an average cell diameter and 3.5 times an average cell diameter. Although the claims of Pat. No. 12,391,914 do not recited the method where a standard deviation of inter-cell spacing between pairs of successive cells is less than 10 µm when measured over 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 pairs of adjacent cells in the first ordered stream of cells as recited in instant claims 17 and 32, would recognize the standard deviation of the spacing between cells would be a matter of routine optimization depending on different factors such as cell size, flow rate, channel dimensions and shape and cell concentration.
Although the claims of Pat. No. 12,391,914 do not recited the method with the claimed ranges of the ratio between the width of microchannel and the diameter of the cells as recited in claims 36-39 and 42-45, the range of the diameter of the cells as recited in claims 40 and 46, or the range of the diameter of the cells as recited in claims 41 and 47, one of ordinary skill in the art would recognize that this ratio, the size of the cells and the width of the channel are all result effective variables and would be matter of routine optimization as evidenced by Amini and dependent on the cell types used. Amini reports that in methods of ordering particles such as cells in microfluidic devices that the closer the size of the channel in is to the bead size or particle size, the faster and more efficient the separating, focusing and ordering (abstract and 0042).
Although the claims of Pat. No. 12,391,914 do not recite that the maximum concentration of the first order stream of cells or the second ordered stream cells is defined by the formal recited in claims 48 and 49, these equations would be readily recognized by one of ordinary skill in the art to be adapted depending on the rate of the fluid movement through the channels to be determine the maximum concentration of cells.
The limitations of instant claims 50 and 51, are recited by claims 1 and 5 of Pat. No. 12,391,914 where generating the single droplet comprises: contacting the flowing first aqueous phase and the second aqueous phase with one another, where the contacting creates a single aqueous phase comprising the first ordered stream of cells and the second ordered stream of cells and where the contacting of the flowing first aqueous phase and the second aqueous phase to create the single aqueous phase occurs at a location at or prior to the junction. The limitations of instant claim 52, are recited by claims 1 and 5 of Pat. No. 12,391,914 where generating the single droplet further comprises: contacting the flowing oil phase with the single aqueous phase to form a cone configuration within the junction, wherein the single droplet is generated at a tip of the cone configuration.
The limitations of instant claims 53-55, are recited by claims 1 and 5 of Pat. No. 12,391,914 where the cells from the first ordered stream of cells and the cells from the second ordered stream of cells are different cells, where the cell from the first ordered stream of cells is a T-cell, and where the cell from the second ordered stream of cells is an antigen presenting cell (APC).
Although the claims of Pat. No. 12,391,914 do not recited the method where the flow rate of the first or second aqueous phase flow rate is between 10-60 µL/min or 45 µL/min, or where the flow rate of the oil phase between 10-60 µL/min or 45 µL/min as recited in instant claims 57-62, would recognize the flow rate for all of the fluids in a method of encapsulating cells in a droplet is a result effective variable and would be matter of routine optimization depending on such factors as channel size, cell size, desired droplet size and rate of production.
The limitations of instant claims 158 and 159, are recited by claim 11 of Pat. No. 12,391,914, where the first microchannel tapers or the second microchannel tapers down from a first width between 40 µm to 100 µm to a second width between 10 µm to 40 µm as the first microchannel approaches the junction.
The limitations of instant claim 160, are recited by claims 5 and 15 of Pat. No. 12,391,914, where the width of the junction is between 40 to 125 µm.
The limitations of instant claim 161, are recited by claim 17 of Pat. No. 12,391,914, where the width of the width of the third channel between 5 to 500 µm.
The limitations of instant claim 162, are recited by claim 5 of Pat. No. 12,391,914, where the method includes flowing the single droplet away from the junction through a nozzle region and flowing the single droplet through a post-nozzle region.
The limitations of instant claim 163, are recited by claim 6 of Pat. No. 12,391,914, where the width of the nozzle is between 10-150 µm and the length of the nozzle is between 20 to 500 µm.
The limitations of instant claim 164, are recited by claim 8 of Pat. No. 12,391,914, where the a width of the post-nozzle region is between 50 pm to 1000 µm.
Claims 1-17, 21-32, 36-49, 53-55, 57-62, and 69 are provisionally rejected on the ground of nonstatutory obviousness-type double patenting as being unpatentable over claims 1-5 and 1-14 of copending Application No. 19/272338. Although the conflicting claims are not identical, they are not patentably distinct from each other because the instant claims encompass those of the issue patent. In addition, both claim methods for encapsulating two cells in a single droplet.
The limitations of instant claims 1 and 69, are recited by claim 1 of Appl. No. 19/272338. Specifically, claim 1 of Appl. No. 19/272338 recites flowing a first aqueous suspension or phase containing a first ordered stream of cells in a first microchannel to a junction; flowing a second aqueous suspension or phase containing a second ordered stream of cells in a second microchannel to a junction; flowing an oil phase in a third microchannel to a junction; and at the junction generating the single droplet formed from the first aqueous phase, the second aqueous phase and the oil drop where the droplet contains a first stream and a second stream cell.
The limitations of instant claim 2, are recited by claim 1 of Appl. No. 19/272338 which recites the fraction of droplets containing both a cell from the first stream and the second stream is higher than what would be predicted based on a Poisson distribution (abstract). Although the claims of Appl. No. 19/272338 do not recited the method wherein the fraction exceeds the predicted fraction by a factor ranging from 2-3 as recited in instant claim 3, one of ordinary skill in the art would recognize that this fraction of cells produced containing one of each cell type is a result effective variable which can be adjusted by controlling different factor in the system such as the flow rates of the fluids in the different microchannels.
The limitations of instant claim 4, are recited by claim 10 of Appl. No. 19/272338 where the method generates single droplets at a rate of at least 5,000 droplets per second. Although the claims of Appl. No. 19/272338 do not recited the method generates single droplets at a rate of at least 8,000 droplets per second as recited in instant claim 5, one of ordinary skill in the art would recognize the rate of droplet formation would be a matter of routine optimization depending on different factors such as the rate at which the individual fluids flow, the composition of the fluids and the desired production rate.
Although the claims of Appl. No. 19/272338 do not recited the method where the cells of the first and second ordered stream of cells are aligned along a central axis or edge of the first and second microchannel as recited in instant claims 6 and 21, or where the cells of the second ordered stream are aligned through inertial focusing while flowing through the channels as recited in instant claims 7 and claim 22, by practicing the method of claim 1 of Appl. No. 19/272338 where the cells are flowed through the undulating regions this would inherently occur.
The limitations of instant claims 8, 12-14, 23, and 27-29, are recited by claims 1 and 11-14 of Appl. No. 19/272338 where the first and second microchannels comprises between 60-120 undulating portions where the portions comprising at least a 90 degree or a 180 degree change in a flow vector across a length of the undulating portion. These undulating portions are curves. Although the claims of Appl. No. 19/272338 do not recited the method where the wherein the curved region is between 150-300 mm in length as recited in instant claims 9 and 24, wherein the curved region is between 50-150 mm in length as recited in instant claims 10 and 25, or wherein the curved region is about 100 mm in length as recited in instant claims 11 and 26, one of ordinary skill in the art would recognize the length of the curved region would be a matter of routine optimization depending on different factors such as the cell size and to control or achieve equilibrium positions in inertial focusing. Similarly, claims 11 and 13 recite between 30-180 undulating portions Although claims 11 and 13 do not teach the exact ranges recited in the instant claims, the ranges overlap significantly with the ranges taught. Furthermore, one of ordinary skill in the art would recognize that the number of undulating portions is a result effective variable and that the number of undulating portions would be matter of routine optimization.
The limitations of instant claims 15, 16, 30 and 31, are recited by claims 2 and 3 of Appl. No. 19/272338 where an inter-cell spacing for at least 80% of cells in the first and second ordered streams is between 1.5 times an average cell diameter and 3.5 times an average cell diameter. The limitations of instant claims 17 and 32, are recited by claims 4 and 5 of Appl. No. 19/272338, where the method where a standard deviation of inter-cell spacing between pairs of successive cells is less than 10 µm when measured over 10 or 20 pairs of adjacent cells in the first ordered stream of cells as recited in instant claims 17 and 32, would recognize the standard deviation of the spacing between cells would be a matter of routine optimization depending on different factors such as cell size, flow rate, channel dimensions and shape and cell concentration.
Although the claims of Appl. No. 19/272338 do not recited the method with the claimed ranges of the ratio between the width of microchannel and the diameter of the cells as recited in claims 36-39 and 42-45, the range of the diameter of the cells as recited in claims 40 and 46, or the range of the diameter of the cells as recited in claims 41 and 47, one of ordinary skill in the art would recognize that this ratio, the size of the cells and the width of the channel are all result effective variables and would be matter of routine optimization as evidenced by Amini and dependent on the cell types used. Amini reports that in methods of ordering particles such as cells in microfluidic devices that the closer the size of the channel in is to the bead size or particle size, the faster and more efficient the separating, focusing and ordering (abstract and 0042).
Although the claims of Appl. No. 19/272338 do not recite that the maximum concentration of the first order stream of cells or the second ordered stream cells is defined by the formal recited in claims 48 and 49, these equations would be readily recognized by one of ordinary skill in the art to be adapted depending on the rate of the fluid movement through the channels to be determine the maximum concentration of cells.
The limitations of instant claims 53-55, are recited by claim 1 of Appl. No. 19/272338 where the cells from the first ordered stream of cells and the cells from the second ordered stream of cells are different cells, where the cell from the first ordered stream of cells is a T-cell, and where the cell from the second ordered stream of cells is an antigen presenting cell (APC).
Although the claims of Appl. No. 19/272338 do not recited the method where the flow rate of the first or second aqueous phase flow rate is between 10-60 µL/min or 45 µL/min, or where the flow rate of the oil phase between 10-60 µL/min or 45 µL/min as recited in instant claims 57-62, would recognize the flow rate for all of the fluids in a method of encapsulating cells in a droplet is a result effective variable and would be matter of routine optimization depending on such factors as channel size, cell size, desired droplet size and rate of production.
Conclusion
No claims are allowed.
Examiner Contact Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to EMILY ANN CORDAS whose telephone number is (571)272-2905. The examiner can normally be reached on M-F 9:00-5:30 EST.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Peter Paras can be reached on 571-272-4517. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of an application may be obtained from the Patent Application Information Retrieval (PAIR) system. Status information for published applications may be obtained from either Private PAIR or Public PAIR. Status information for unpublished applications is available through Private PAIR only. For more information about the PAIR system, see http://pair-direct.uspto.gov. Should you have questions on access to the Private PAIR system, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative or access to the automated information system, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/EMILY A CORDAS/Primary Examiner, Art Unit 1632
Prosecution Insights