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 .
Priority
This application, Serial No. 17/495,133 was filed on 06 October 2021 and claims foreign priority under 35 U.S.C. 119(b) and 37 CFR 1.55 to Korean Patent Application No. KR10-2020-0129915, filed on 08 October 2020, Korean Patent Application No. KR10-2021-0024191, filed on 23 February 2021, and Korean Patent Application No. KR10-2021-0094785, filed on 20 July 2021; the earliest effective filing date is 08 October 2020.
Status of the Claims
Claims 1-6, 10- 12 and 16 are cancelled. Claims 7-9, 13-15, 17-21 are pending. Claim 20 is withdrawn. Claims 7-9, 13-15, 17-19 and 21 are examined herein.
Withdrawn Rejections/Objections
The rejection of claims 7-9, 11-15, 17-19, and 21 under 35 U.S.C. 112(b) is withdrawn in response to Applicant’s amendment.
The rejection of claims 7-9, 11-15, 17-19, and 21 under 35 U.S.C. 103 is withdrawn in response to Applicant’s amendment.
New 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 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 7-9, 13-15, 17-19, and 21 are rejected under 35 U.S.C. 103 as being unpatentable over Fogelstrand et al (US Patent No.: 11913961 B2) in view of BIO RAD (Secondary Antibodies Guide: Secondary Antibodies, Making Your Secondary Detection Work for You; 07/19), Geiser et al (US Patent No.: 5804393), and Onuallain et al (WO 2009/048631 A1) as evidenced by Thermo (Pierce Centrifuge Columns, 0.8mL Instructions, 2007).
With respect to claim 7, Fogelstrand teaches throughout the publication various embodiments for preparing biological samples to use in an immunolabeling process. Specifically, Fogelstrand teaches that an indirect immunolabeling method may be used for signal amplification (column 1, lines 64-65), and involves reporter molecule of choice, a primary antibody bound to a target molecule (target protein) in a sample and a secondary antibody bound to the first antibody with the option of binding several second antibodies to each primary antibody for increased signal amplification (column 1, lines 65- column 2 line 3) in which the reporter molecule may be fluorescent (column 1, lines 47-49); Fogelstrand discloses embodiments to include multiple target proteins (FIG.3, column 7, lines 38-67), and further discloses that a negative effect of indirect immunolabeling includes a rise in unspecific cross-binding reactions between secondary antibodies to endogenous antibodies, between same primary antibody classes (species) as commercially available species are limited, as well as secondary antibodies cross-reacting to primary antibodies of the same class (species)- particularly when performing multi-immunolabeling in a sample (column 2, lines 1-50). Further, Fogelstrand specifies that a signal enhancer system for immunolabeling enables an unlimited number of amplification steps when cross reactions are not present in such an amplification scheme, to include possible cross reaction sources present in the biological sample, sample processing reagents or staining reagents (column 2, lines 63-67 and column 3, lines 1-11). Fogelstrand discloses, “… order of the steps may differ from what is depicted… steps may be performed concurrently or with partial concurrence. Such variation will depend on designer choice. All such variations are within the scope of the disclosure… even though the invention has been described with reference to specific exemplifying embodiments thereof, many different alterations, modifications and the like will become apparent for those skilled in the art; emphasis by examiner. This reads on fluorophore conjugation before and after the removal of cross-reactivity. The step of adding the fluorophore to the secondary antibodies at various points of the method of processing the secondary antibodies is considered to be prima facie obvious in the absence of new or unexpected results as a consequence of the particularly claimed order (See MPEP 2144.04(IV)(C) and See also In re Burhans, 154 F.2d 690, 69 USPQ 330 (CCPA 1946) (selection of any order of performing process steps is prima facie obvious in the absence of new or unexpected results).
Fogelstrand differs from the instant claim in failing to specifically teach the use of “prepared” antibodies and secondary antibodies in an amplification scheme as defined by the instant claim set, wherein a “prepared antibody” is defined as an antibody of choice, the antibody to be purified, which has had all undesired cross reactions with different antibodies removed in a single step by specifically using an NHS-activated agarose gel by performing a centrifugation process in the absence of an elution process.
However, BIO RAD teaches throughout the publication the intricacies of choosing secondary antibodies as applicable to certain applications and when additional purification steps are required to remove undesired cross-reactions, known as cross-adsorption (pg.5, Select Your Secondary Antibody, Tips; pg. 8, #5). Specifically, BIO RAD teaches multiplexing as an example application where the use of differentially labeled primary antibodies may be achieved by the careful choice of antibodies based on different host/target species, different antibody classes, and/or different antibody isotypes, noting that “…when mixing multiple secondary antibodies, ensure they are cross-adsorbed against unwanted targets to ensure specificity of your staining…” (pg.9, #9; ch.8, Multiplexing, pg.33-34). Additionally, BIO RAD discloses that cross-adsorbed secondary antibodies are generated by the use of affinity chromatography [AC] to bind undesired antibodies, which include antibodies from offending cross-reactive species, antibodies of a different class or antibodies of a different isotype, wherein, “…the unbound pool of antibodies is then considered cross-adsorbed against the immobilized antigen and should show no reactivity towards it. Efficient cross-adsorption may require multiple passes through a column and cross-adsorption against multiple antigens will require passes through multiple columns.” (pg.22, ¶3); emphasis by examiner. As evidenced in Thermo, the process of removing unwanted components by affinity chromatography is regularly performed with a centrifugation process in the absence of an elution step (Thermo, p. 1, step 4). BIO RAD is silent on teaching the specific use of agarose gel in the disclosed AC technique used when removing undesired cross-reactions between antibodies. See also Chapter 6, page 21.
Fogelstrand in view of BIO RAD does not teach that the processed secondary antibodies have had all undesired cross reactions with different antibodies removed in a single step by specifically using an NHS-activated agarose gel by performing a centrifugation process in the absence of an elution process.
However, Geiser teaches that, when processing cross-reaction-free antibodies, the target antibodies can be processed through an agarose-based column loaded with a plurality of cross-reacting ligands, (in the case of Geiser, a plurality of toxins), in order to remove cross-reactive antibodies to those ligands (col. 4, lines 35-44). Geiser further teaches that, consistent with the teachings of BIO RAD, the target antibodies lacking cross-reactivity to the ligands will be found in the flow-through, i.e. not eluted from the column (col. 4, lines 35-44).
It would have been obvious to one of skill in the art before the effective filing date of the claimed invention to modify the method of preparing the cross-reaction-free antibodies taught by Fogelstrand in view of BIO RAD by using an agarose-based column comprising a plurality of cross-reactive ligands as taught by Geiser, i.e. the multiple orthogonal antibodies taught by Fogelstrand. An artisan would have been motivated to use an agarose-based column comprising a plurality of cross-reactive ligands as taught by Geiser in order to reduce the number of steps necessary to prepare the cross-reaction-free antibodies compared to the multiple columns taught by BIO RAD. An artisan would have had a reasonable expectation in using an agarose-based column comprising a plurality of cross-reactive ligands as taught by Geiser because, as further taught by Geiser, these columns can effectively reduce cross-reactive antibodies from a pool of antibodies, which is the goal of the method taught by Fogelstrand in view of BIO RAD.
Fogelstrand in view of BIO RAD and Geiser does not teach that the agarose gel is an NHS-activated agarose gel or that the NHS-activated agarose gel is treated with a trisaminomethane solution after connecting the cross-reaction ligands.
However, Onuallain teaches a method of preparing an NHS-based agarose gel column with ligands (para. 00213, lines 1-14). Onuallain teaches having NHS groups on the surface of the agarose gel to react with amine groups on the ligands, then treating the gel with trisaminomethane, i.e. Tris-HCl, to terminate the coupling reaction, i.e. remove the remaining NHS groups (para. 00213, lines 1-14).
It would have been obvious to one of skill in the art before the effective filing date of the claimed invention to make the ligand-bound agarose gel taught by Fogelstrand in view of BIO RAD and Geiser using the NHS-based method taught by Onuallain. Doing so is considered to be Combining Prior Art Elements According to Known Methods To Yield Predictable Results (See MPEP 2143(I)(A)). An artisan wanting to use a ligand-bound agarose gel as taught by Fogelstrand in view of BIO RAD and Geiser would recognize that the NHS-activated method taught by Onuallain would predictably generate such a gel, as the function of the NHS-activated method taught by Onuallain performs the function of making such a ligand-bound agarose gel.
Regarding claim 8, Fogelstrand teaches antibodies conjugated with reporter molecules (column 1, lines 42-44), and specifies that reporter molecules are, “… typically selected from a group comprising a fluorochrome, an enzyme, a peptide, quantum dots, and a transition metal. Other known or future reporter molecules are possible and within the scope of the invention… reporter molecule(s) are typically used in a subsequent detection/analysis process, such as for example by illumination of the biological sample under a microscope to detect a light from a fluorochrome” emphasis by examiner (column 4, lines 33-41); this reads on the instant claim because Fogelstrand’s listed reporter molecules are non-limiting and substituting a fluorophore in place of a fluorochrome would have been obvious.
Regarding claim 9, in light of Fogelstrand and BIO RAD as applied above, Geiser teaches removing cross-reactions from antibodies while purifying said antibodies in the flow-through. For this reason, one of ordinary skill in the art would have been motivated to use antibodies as ligands because cross-adsorption technique may use antibodies of different species, class, or isotype as the ligand and the antigen in gel, therein removing cross-reactions while simultaneously purifying the antibody components that will be collected in the flow-through.
Regarding claims 13-15 and 17-18, in light of BIO RAD and Geiser it is reiterated that Fogelstrand teaches the same general component set-up for immunolabeled signal enhancing systems involving antibody manipulation which includes a target, reporter molecule(s), a primary antibody, and the desired amount of enhancing “chain” secondary antibodies wherein the total number of amplifying secondary antibody layers is dependent on user design, desired outcome, and reducing/eliminating cross-reactions when performing multi-immunolabeling (see claim 7 analysis herein), to include scheme components in complexes of primary and secondary antibodies (column 3, lines 27-30), as well as slight variations to order of steps (instant claim 15), singularity/plurality assigned, and included components (column 8, lines 45-60). This reads on instant claims 13 which recites a different [prepared] antibody pair comprising a first secondary antibody and a second secondary antibody, because the distinction of two secondary antibody components in the amplifying scheme (regardless of where they are present in terms of layer/level from the target) by labeling it a pair, does not diverge from the scope of Fogelstrand’s teaching; accordingly, the binding order of antibodies within an amplification scheme, as well as whether bound in pair, singular, or in layer format and number of times a step is repeatedly performed for increased amplification does not depart from Fogelstrand’s teachings. Fogelstrand further teaches the amplification layers created through a possible “… unlimited number of amplification steps” as disclosed in claims 14 and 17 (column 2, lines 63-67), as well as a fluorescence signal enhancing system achieved by the binding of a primary antibody to its respective target protein, the binding of a [prepared] first secondary antibody to its respective primary antibody, and the binding of a [prepared] second secondary antibody to its respective first secondary antibody as recited in instant claims 15 and 18 (column 1, lines 65- column 2 line 3).
Regarding claim 19, in light of BIO RAD and Geiser, Fogelstrand teaches the disclosed host/target relationship described wherein “…a target of the first secondary antibody is identical with a host of the second secondary antibody; and a host of the first secondary antibodies is identical with a target of the second secondary antibody”, by reciting, “The secondary antibody is usually raised against the immunoglobulin class of the animal species in which the primary antibody has been raised…” (column 2 lines 3-8). Since Fogelstrand makes clear that amplification steps may be unlimited as long as cross-reactions are removed (column 8, lines 27-44), it would have been obvious to repeat the disclosed species affinity logic between any subsequent secondary antibody layers to increase signal amplification further.
Regarding claim 21, BIO RAD outlines the relationship between affinity chromatography and creating secondary antibodies of interest to reduce unwanted cross reactions and discloses a variety of different secondary antibodies (pg. 6, Secondary Antibodies at a Glance) as well as multiplexing using a combination of different host species (pg. 33, last ¶). Therefore, a skilled artisan would have had a reasonable expectation of success in modifying the method of Fogelstrand by using the agarose gel column taught by Geiser to purify antibodies by cross-absorbing different populations of antibodies as taught by BIORAD because BIO RAD teaches throughout the publication the intricacies of choosing secondary antibodies as applicable to certain applications and when additional purification steps are required to remove undesired cross-reactions and their method is generic to different population of antibodies by from different species.
Response to Arguments
Applicant’s arguments with respect to claim(s) 7-9, 11-15, 17-19, and 21 have been considered but are moot because the new ground of rejection does not rely on any reference applied in the prior rejection of record for any teaching or matter specifically challenged in the argument.
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.
/C.E./Examiner, Art Unit 1677
/BAO-THUY L NGUYEN/Supervisory Patent Examiner, Art Unit 1677 September 23, 2026
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