CTNF 17/721,558 CTNF 95271 DETAILED ACTION Notice of Pre-AIA or AIA Status 07-03-aia AIA 15-10-aia The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA. Status of the Application The Amendment filed February 3, 2026 is acknowledged. Claims 1-6, 8-11, 14 and 16-17 were pending. Claims 1, 4-6, 8-11, 14, 16-17 and new claims 18-20 and 25-28 are being examined on the merits. Claims 2-3 are canceled. New claims 21-24 are newly withdrawn. Response to Arguments Applicant’s arguments filed February 3, 2026 have been fully considered. The following objections and rejections are WITHDRAWN in view of Applicant’s arguments and the instant amendments to the specification and claims: Objection to the specification – title Prior art rejections of claim 1 and its dependent claims The following rejections are MODIFIED in view of Applicant’s arguments: Prior art rejection of claim 11 Response to arguments regarding prior art rejections Claim 1 and dependent claims Applicant argues that the prior art rejections of claim 1 should be withdrawn because the cited art does not teach or suggest all of the limitations of instantly amended claim 1, including, for example, incorporating the alkaline phosphatase enzyme as the label in the detection complex (Remarks, pp. 15-17). The Examiner agrees, at least, that the cited art does not teach or suggest incorporating the alkaline phosphatase enzyme as the label in the detection complex. The prior art rejections based on Ikebukuro are withdrawn. However, in view of an additional search, new prior art rejections are added below. Claim 11 Applicant argues that the prior art rejection of claim 11 is unreasonable and should be withdrawn because … PNG media_image1.png 110 936 media_image1.png Greyscale PNG media_image2.png 229 821 media_image2.png Greyscale (Remarks, pp. 18-19). The Examiner agrees in part and disagrees in part. First, regarding Applicant’s statement that “[a] conjugate solution configured for measurement … with an … ISFET requires connecting the conjugate solution to an … ISFET …”, the Examiner notes that this is a method step (i.e., “connecting …”). However, claim 11 is product claim and does not comprise method steps. However, as noted below in the Claim Interpretation section, the limitation reciting that the conjugate solution is configured for measurement by an ISFET at least requires that the conjugate solution is capable of generating a phenomenon that is detectable by an ISFET, e.g., changes in hydrogen ion concentration (see Jaung, Fig. 1; cited below in the prior art rejections). Further, if a conjugate solution is attached to a detection system that is not an ISFET, then it does not appear to be reasonable to interpret that teaching as meeting the limitation of being configured for measurement by an ISFET. Thus, the teaching in Ikebukuro that the conjugate solution is attached to a device that is not an ISFET, does not, in and of itself, read on the “configured for measurement …” limitation. Rather, the Ikebukuro assay and corresponding components would have to be modified to substitute in an ISFET detection system in order to teach or suggest that limitation. The prior art rejection of claim 11 is modified in view of the above comments. Election/Restrictions 08-04 AIA Newly submitted claim s 21-24 are directed to an invention that is independent or distinct from the invention originally claimed for the following reasons: the subject matter of the group of independent claims 1 and 11 and the group of claims 21-24 are related as product and process of use. The inventions can be shown to be distinct if either or both of the following can be shown: (1) the process for using the product as claimed can be practiced with another materially different product or (2) the product as claimed can be used in a materially different process of using that product. See MPEP § 806.05(h). In the instant case, the product of claims 1 and 11 can be used in a materially different process, e.g., by using a different detection technique, such as a potentiometric electrochemical technique using a glass pH electrode . Since applicant has received an action on the merits for the originally presented invention, this invention has been constructively elected by original presentation for prosecution on the merits. Accordingly, claim s 21-24 are withdrawn from consideration as being directed to a non-elected invention. See 37 CFR 1.142(b) and MPEP § 821.03. To preserve a right to petition, the reply to this action must distinctly and specifically point out supposed errors in the restriction requirement. Otherwise, the election shall be treated as a final election without traverse. Traversal must be timely. Failure to timely traverse the requirement will result in the loss of right to petition under 37 CFR 1.144. If claims are subsequently added, applicant must indicate which of the subsequently added claims are readable upon the elected invention. Should applicant traverse on the ground that the inventions are not patentably distinct, applicant should submit evidence or identify such evidence now of record showing the inventions to be obvious variants or clearly admit on the record that this is the case. In either instance, if the examiner finds one of the inventions unpatentable over the prior art, the evidence or admission may be used in a rejection under 35 U.S.C. 103 or pre-AIA 35 U.S.C. 103(a) of the other invention. 07-30-03-h AIA Claim Interpretation Claims 11 and 18 each recite the limitation “the conjugate solution is configured for measurement of the detectable phenomenon with an [ISFET]”. An ISFET is an electrochemical detection device which can be used to measure changes in ion concentrations in solution. An example of an ISFET for detecting changes in proton concentrations in solution is shown in Juang Fig. 1 (cited below in conjunction with the prior art rejections). Thus, the “configured for measurement …” limitation is being interpreted as requiring, in part, that the conjugate solution generates, e.g., a change in hydrogen ion concentrations. In addition, if a conjugate solution is attached to a detection system that is not an ISFET, then it does not appear to be reasonable to interpret that teaching alone as meeting the limitation of being configured for measurement by an ISFET. Thus, the “configured for measurement …” limitation is being construed as requiring both that the conjugate solution is capable of generating a phenomenon which can be detected by an ISFET, and that the conjugate solution is not attached to a detection device that is not an ISFET. Claim Rejections - 35 USC § 112 07-30-02 AIA 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. 07-34-01 Claim 6 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. Lack of antecedent basis rejections Claim 6 recites the limitations "the chemical substituent" in l. 4 and “the nucleic acid binding protein” in l. 8. There is insufficient antecedent basis for each of these limitations in the claim. Claim 1, from which claim 6 depends, recites two “chemical substituents” – one for the binding substance and one for the label, and two “nucleic acid binding proteins” – one for the binding substance and one for the label. It is not clear which of each of these is being referred to in the claim 6 reference to “chemical substituent” and “nucleic acid binding protein”. 07-36 AIA The following is a quotation of 35 U.S.C. 112(d): (d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers. The following is a quotation of pre-AIA 35 U.S.C. 112, fourth paragraph: Subject to the following paragraph [i.e., the fifth paragraph of pre-AIA 35 U.S.C. 112], a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers. Claims 26 and 28 are rejected under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends. Claim 26 recites the limitation “the greater an amount of the target substance per unit volume in the sample, the greater an amount of change in pH caused by the phenomenon”. As noted below in the prior art rejections, such a configuration is a property of the claim 25 system. Since claim 26 does not recite any additional structural limitations to the claim 25 system, it does not further limit the claim 25 system and is in improper dependent form. Claim 28 similarly recites a property of the claim 27 system, and consequently claim 28 is rejected with reasoning corresponding to the claim 26 rejection. Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements. Claim Rejections - 35 USC § 103 07-20-aia AIA 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. 07-20-02-aia AIA 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. 07-21-aia AIA Claim s 1, 4-6, 8, 16 and 18-20 are rejected under 35 U.S.C. 103 as being unpatentable over Marshall (US Patent App. Pub. No. 2016/0223530) in view of Centi (Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich Assay, Electroanalysis , 21(11): 1309-1315, 2009), as evidenced by Hauer (WO 01/07573 A1; with English translation) and Alfa Chemistry (Understanding ALP Substrates: pNPP and BCIP/NBT in Biochemical Diagnostics, 2026) . Regarding independent claim 1 and dependent claims 4-6, 8 and 19-20 , Marshall teaches … A conjugate solution comprising: conjugates each having a binding substance fused with a label and configured to bind to a target substance, optionally a protein, nucleic acid, lipid or carbohydrate, and exhibit a detectable phenomenon of a change in concentration of an ion, optionally by the generation of a hydrogen ion when pNPP is the substrate of the label, the binding substance having an activity to bind to the target substance and being configured to form a target-substance-bound conjugate within a solution containing the conjugate and the target substance, and the conjugate solution containing target-substance-bound conjugates without any unbound conjugates, the label of the target-substance-bound conjugate being activated in the conjugate solution causing the detectable phenomenon to occur (claims 1, 14: a solution comprising a target substance and a reporter enzyme detection probe (i.e., a binding substance fused with a label), wherein the detection probe is configured to bind specifically with the target, and wherein a corresponding enzyme substrate is present which is capable of generating an ionizable product (i.e. which changes the concentration of an ion in solution) – further, the solid phase is washed to remove unbound conjugates, which are thus not present in the solution – further, the reporter enzyme detection probe comprises an aptamer and the reporter enzyme is alkaline phosphatase; also Fig. 1; paras. 31, 65, 157, 168, 225, 299); wherein the binding substances is a nucleic acid aptamer, the binding substance having a chemical substituent, optionally biotin (claims 1, 14; Fig. 1; paras. 19, 157, 168, 225, 405 – binding substance is biotinylated); the label is an alkaline phosphatase with a chemical substituent, optionally streptavidin (claims 1, 14; Fig. 1; paras. 19, 157, 168, 225, 405 – the label is conjugated to streptavidin); and the chemical substituent of the binding substance is conjugated with the chemical substituent of the label (claims 1, 14; Fig. 1; paras. 19, 157, 168, 225, 405 – the biotin is bound to the streptavidin). Marshall does not teach any particular length for the aptamer, however, Centi teaches that the aptamer has 10-100 bases (section 2.2: 44 nucleotides). Further, regarding the limitations relating to exhibiting a detectable phenomenon of a change in concentration of an ion, optionally by the generation of a hydrogen ion, these limitations are inherent properties of using alkaline phosphatase in the presence of pNPP at an alkaline pH, as evidenced by Alfa Chemistry and Hauer. Specifically, Alfa Chemistry teaches that pNPP is catalyzed to p-nitrophenol (p. 2, bottom para.), while Hauer teaches that, at pH 8.1-8.2, p-nitrophenol is deprotonated thus producing the yellow color which can be used for colorimetric detection (p. 114, para. 2). The deprotonation thus generates hydrogen ions in solution. Also see, MPEP 2112.01 which states that “when the structure recited in the references is substantially identical to that of the claims, claimed properties or functions are presumed to be inherent”. Prior to the effective filing date of the instant invention, it would have been prima facie obvious to modify the Marshall binding substance with the aptamer length of Centi. The ordinary artisan would have been motivated to do so to customize the assay as desired through routine optimization, and because aptamers of that length are recognized in the art as being suitable for that the intended purpose of being used in biosensor sandwich assays (e.g., Centi, Fig. 1). See MPEP 2144.07. The ordinary artisan would have had an expectation of success as the design and modification of nucleic acid assays is well-known the art. Regarding dependent claim 16 , Marshall teaches that the binding substance is bound to the target substance and that the target substance is not itself attached to/ immobilized on a solid phase (e.g., Fig. 1) . 07-22-aia AIA Claim 9 is rejected under 35 U.S.C. 103 as being unpatentable over Marshall (US Patent App. Pub. No. 2016/0223530) in view of Centi (Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich Assay, Electroanalysis , 21(11): 1309-1315, 2009), as evidenced by Hauer (WO 01/07573 A1; with English translation) and Alfa Chemistry (Understanding ALP Substrates: pNPP and BCIP/NBT in Biochemical Diagnostics, 2026) as applied to claim 1 above, and further in view of Sullenger 1 (US Patent App. No. 2020/0095636 A1) . Regarding dependent claim 9 , Sullenger teaches that the chemical substituent is N-hydroxysuccinimide (para. 55). Prior to the effective filing date of the instant invention, it would have been prima facie obvious to substitute the Sullenger substituent into the modified Marshall composition, discussed above. Sullenger teaches that such chemical substituents are useful for attaching labels to aptamers. The selection of a known material based on its suitability for its intended purpose supports a prima facie obviousness determination. MPEP 2144.07. Further, the ordinary artisan would have been motivated to optimize the chemical substituents through routine experimentation to customize the assay as needed. The ordinary artisan would have had an expectation of success as attaching various kinds of labeling moieties to nucleic acids is well-known in the art . 07-21-aia AIA Claim s 14 and 17 are rejected under 35 U.S.C. 103 as being unpatentable over Marshall (US Patent App. Pub. No. 2016/0223530) in view of Centi (Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich Assay, Electroanalysis , 21(11): 1309-1315, 2009), as evidenced by Hauer (WO 01/07573 A1; with English translation) and Alfa Chemistry (Understanding ALP Substrates: pNPP and BCIP/NBT in Biochemical Diagnostics, 2026) as applied to claim 1, and further in view of Samson 2 (Biosensors: Frontiers in rapid detection of COVID-19 , 3 Biotech , 10:385, 1-9, 2020) . Regarding dependent claim 14 , Samson teaches the use of aptamer-based biosensors to detect SARS-CoV-2 (e.g., abstract). Regarding dependent claim 17 , Marshall teaches that the binding substance is an aptamer (e.g., claims 1, 14; para. 168). Further, Samson teaches that the binding substance is configured to bind to the target substance without the target substance being separate (p. 2, left col., para. 1: detect intact viral particles), and teaches that the target substance is an envelope protein of SARS-CoV-2 (Fig. 1). Prior to the effective filing date of the instant invention, it would have been prima facie obvious to further modify the modified Marshall composition, discussed above, to detect SARS-CoV-2. Marshall teaches that the composition is useful for detecting a variety of targets, including viruses (paras. 299) and has high-sensitivity. Samson teaches that SARS-CoV-2 can be detected with aptamer-based biosensors. The ordinary artisan would have been motivated to try to make the composition capable of detecting the SARS-CoV-2 target with the Marshall assay, with the expectation that doing so would result in the advantage of a highly sensitive assay for detecting SARS-CoV-2. The ordinary artisan would have also been motivated to try to make the composition capable of detecting the SARS-CoV-2 envelope protein without the envelope protein being separated from the virus particle with the expectation that doing so would result in the advantage of an efficient and lower cost assay, as purification steps could be omitted. The ordinary artisan would have had an expectation of success as the design of compositions that are capable of detecting proteins from a variety of organisms is well-known in the art . 07-22-aia AIA Claim s 6 and 10 are rejected under 35 U.S.C. 103 as being unpatentable over Marshall (US Patent App. Pub. No. 2016/0223530) in view of Centi (Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich Assay, Electroanalysis , 21(11): 1309-1315, 2009), as evidenced by Hauer (WO 01/07573 A1; with English translation) and Alfa Chemistry (Understanding ALP Substrates: pNPP and BCIP/NBT in Biochemical Diagnostics, 2026) as applied to claim 1 above, and further in view of Ma 3 (Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow, Nature Biotechnology , 34(5): 528-531, 2016) . Regarding dependent claims 6 and 10, Marshall teaches that the binding substance and the label are fused with each other (as noted above in claim 1), while Ma teaches that the fusion is through a nucleic acid binding protein, specifically a CRISPR complex (abstract: system based on dCas9 combined with sgRNA; Fig. 1: sgRNAs are bound to labels; p. 528, left col., para. 2: discusses fusing sgRNAs to aptamers; thus, sgRNA can be used to fuse together aptamers and labels). Prior to the effective filing date of the instant invention, it would have been prima facie obvious to incorporate the Ma sgRNA linker into the Marshall composition, discussed above. Ma teaches that sgRNA is useful for attaching labels to aptamers. The selection of a known material based on its suitability for its intended purpose supports a prima facie obviousness determination. MPEP 2144.07. Further, the ordinary artisan would have been motivated to optimize the component linkages through routine experimentation to customize the assay as needed. The ordinary artisan would have had an expectation of success as attaching various kinds of labeling moieties to nucleic acids is well-known in the art . 07-21-aia AIA Claim 18 is rejected under 35 U.S.C. 103 as being unpatentable over Marshall (US Patent App. Pub. No. 2016/0223530) in view of Centi (Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich Assay, Electroanalysis , 21(11): 1309-1315, 2009), as evidenced by Hauer (WO 01/07573 A1; with English translation) and Alfa Chemistry (Understanding ALP Substrates: pNPP and BCIP/NBT in Biochemical Diagnostics, 2026) as applied to claim 1, and further in view of Pachauri (Biologically sensitive field-effect transistors: from ISFETs to NanoFETs, Essays Biochem , 60(1): 80-90, 2016) and Juang (Proton-ELISA: Electrochemical immunoassay on a dual-gated ISFET array, Biosensors and Bioelectronics , 117, 175-182, 2018) . Regarding dependent claim 18 , regarding the limitation requiring that the conjugate solution is configured for measurement of the detectable phenomenon with an ion-sensitive field effect transistor (ISFET), as noted, Marshall teaches a biosensor sandwich assay which generates hydrogen ions, and also teaches that the aptamers are attached to an detection device (Fig. 1), although the device is not an ISFET. Juang teaches a biosensor sandwich assay that generates hydrogen ions (Fig. 1), and also teaches that ISFETs are capable of detecting changes in solutions due to fluctuations resulting from changes in ion concentration, e.g., hydrogen ion concentration and pH (Fig. 1). Pachauri teaches that ISFETs have certain advantages compared to other electrochemical detection techniques when used in biosensor assays, including miniaturization and high sensitivity, both of which are useful in point-of-care devices (p. 85, last para. to p. 86, first para.). Prior to the effective filing date of the instant invention, it would have been prima facie obvious to modify the Marshall assay and the corresponding conjugate solution to incorporate the Juang ISFET and the corresponding conjugate solution that is configured for measurement by an ISFET. The ordinary artisan would have been motivated to do so to achieve the expected advantages of a conjugate solution which is capable of being used in a miniaturized POC device and which is capable of being use in a highly sensitive assay, as taught by Pachauri. The ordinary artisan would have had an expectation of success as ISFETs are recognized in the art as being useful for detecting conjugate solutions which generate hydrogen ion concentrations, and because the design and modification of nucleic acid assays is well-known in the art . 07-21-aia AIA Claim s 25-26 are rejected under 35 U.S.C. 103 as being unpatentable over Marshall (US Patent App. Pub. No. 2016/0223530) in view of Centi (Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich Assay, Electroanalysis , 21(11): 1309- 1315, 2009), Samson 4 (Biosensors: Frontiers in rapid detection of COVID-19 , 3 Biotech , 10:385, 1-9, 2020) and Lu 5 (US Patent App. Pub. No. 2015/0031014), and as evidenced by Hauer (WO 01/07573 A1; with English translation) and Alfa Chemistry (Understanding ALP Substrates: pNPP and BCIP/NBT in Biochemical Diagnostics, 2026) . Regarding independent claim 25 , Marshall teaches … A system for detecting a target substance, the system comprising: a sample containing conjugates, each conjugate including a binding substance and label conjugated with the binding substance; wherein the binding substance is a nucleic acid aptamer with biotin having activity to bind to the target substance; the label is alkaline phosphatase with streptavidin configured to cause a detectable phenomenon, and the biotin of the binding substance is conjugated with the streptavidin of the label (claims 1, 14: a sample solution comprising a target substance and a reporter enzyme detection probe (i.e., a binding substance fused with a label), wherein the detection probe is configured to bind specifically with the target, and wherein a corresponding enzyme substrate is present which is capable of generating an ionizable product (i.e. which changes the concentration of an ion in solution) – further, the solid phase is washed to remove unbound conjugates, which are thus not present in the solution – further, the reporter enzyme detection probe comprises an aptamer with biotin and the reporter enzyme is alkaline phosphatase; also Fig. 1; paras. 19, 31, 65, 157, 168, 225, 299, 405 – binding substance is biotinylated and the label is conjugated to streptavidin); the detectable phenomenon is generation of hydrogen ion in the sample, which is caused by metabolic activity using disodium 4-nitrophenyl phosphate hexahydrate as a substrate, induced by enzymatic activity of the alkaline phosphatase (paras. 31, 91, 157, 164: the generation of a hydrogen ion when a salt of pNPP is the substrate of the label; as evidenced by Alfa Labs, which teaches that pNPP is catalyzed to p-nitrophenol, while Hauer teaches that, at pH 8.1-8.2, p-nitrophenol is deprotonated thus producing the yellow color which can be used for colorimetric detection – the deprotonation thus generates hydrogen ions in solution). Marshall does not teach any particular length for the aptamer, however, Centi teaches that the aptamer has 10-100 bases (section 2.2: 44 nucleotides). Samson teaches the use of aptamer-based biosensors to detect SARS-CoV-2, specifically the spike glycoprotein (e.g., abstract, Fig. 1). Lu teaches using an enzyme to convert a substrate into a product plus an acid, thus acidifying the solution, and where the acidification is detected with a pH meter which is configured to detect the phenomenon (Figs. 5A-C, 9, 10A-B, 12A-12B, 18; paras. 3, 7-9, 74, 90, 93, 108-117, 118-120: the label is Gox, which can convert glucose into hydrogen peroxide and gluconic acid, thus acidifying the solution – this can be detected with a pH meter). Prior to the effective filing date of the instant invention, it would have been prima facie obvious to modify the Marshall binding substance with the aptamer length of Centi. The ordinary artisan would have been motivated to do so to customize the assay as desired through routine optimization, and because aptamers of that length are recognized in the art as being suitable for the intended purpose of being used in biosensor sandwich assays (e.g., Centi, Fig. 1). See MPEP 2144.07. The ordinary artisan would have had an expectation of success as the design and modification of nucleic acid assays is well-known the art. It would have been further obvious to use the modified Marshall composition to detect the SARS-CoV-2. Marshall teaches that the composition is useful for detecting a variety of proteins and is highly sensitive. Samson teaches that the spike protein of SARS-CoV-2 can be detected with aptamer-based biosensors. The ordinary artisan would have been motivated to try to make the composition capable of detecting the SARS-CoV-2 spike protein target with the Marshall assay, with the expectation that doing so would result in the advantage of a highly sensitive assay for detecting SARS-CoV-2. The ordinary artisan would have had an expectation of success as the design of compositions that are capable of detecting targets from a variety of organisms is well-known in the art. Finally, it would have been additionally obvious to incorporate a pH meter into the system with the Marshall composition. The ordinary artisan would have been motivated to do so to customize the assay as desired through routine optimization, and the use of pH meters is recognized in the art as being suitable for the intended purpose of measuring changes in hydrogen ion concentrations. See MPEP 2144.07. The ordinary artisan would have had an expectation of success as the design and modification of detection systems for nucleic acid assays is well-known the art. Regarding dependent claim 26 , the ordinary artisan would understand that it is an inherent property of the Marshall pNPP plus alkaline phosphatase detection system that the greater the amount of target substance in the sample, the greater amount of hydrogen ions generated and hence the greater the pH change, as evidenced by Alfa Chemistry and Hauer. Alfa Chemistry teaches that the amount of p-nitrophenol produced by alkaline phosphatase cleavage of pNPP is proportional to the amount of target in the sample, thus allowing for quantitative detection of the target (p. 4, Clinical Diagnostics section). Hauer, teaches that the same reaction produces hydrogen ions (p. 114, para. 2). Thus, the ordinary artisan would understand that greater the amount of target substance in the sample, the greater the amount of both p-nitrophenol and hydrogen ions are generated by the alkaline phosphatase reaction . 07-22-aia AIA Claim s 27-28 are rejected under 35 U.S.C. 103 as being unpatentable over Marshall (US Patent App. Pub. No. 2016/0223530) in view of Centi (Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich Assay, Electroanalysis , 21(11): 1309-1315, 2009), Samson (Biosensors: Frontiers in rapid detection of COVID-19 , 3 Biotech , 10:385, 1-9, 2020) and Lu (US Patent App. Pub. No. 2015/0031014), and as evidenced by Hauer (WO 01/07573 A1; with English translation) and Alfa Chemistry (Understanding ALP Substrates: pNPP and BCIP/NBT in Biochemical Diagnostics, 2026) as applied to claim 25 above, and further in view of Sigma (p-Nitrophenyl Phosphate Liquid Substrate System, 2019) . Regarding dependent claim 27 , Sigma, as evidenced by Hauer or Alfa Chemistry, teach that the phenomenon is caused by replacing solution of the sample with another solution in which disodium-4-nitrophenyl phosphate hexahydrate is dissolved to generate the colorimetric reaction. Specifically, Sigma teaches replacing the sample solution with a substrate solution (Procedure, steps 1 through 3), while Hauer teaches that the colorimetric reaction requires a pH of at least 8.1-8.2 (p. 114, para. 2), while Alfa Chemistry teaches a pH of 9.2-9.8 (p. 6, Table, top row). Thus, the ordinary artisan would understand that the Sigma reaction would inherently require a pH of at least 8 in order to generate the recited reaction. Prior to the effective filing date of the instant invention, it would have been prima facie obvious to further modify the modified Marshall system, discussed above, so that the substrate of label is included in the system. The ordinary artisan would have been motivated to do so to customize the system as desired through routine optimization, and would have had an expectation of success as the design and modification of nucleic acid assays and corresponding systems is well-known the art. Regarding dependent claim 28 , the ordinary artisan would understand that it is an inherent property of the Marshall pNPP plus alkaline phosphatase detection system that the amount of change in pH increases over time after replacement with the other (i.e., substrate) solution, as evidenced by Alfa Chemistry and Hauer. Alfa Chemistry teaches that the amount of change in absorbance increases over time after the solution replacement (Fig. 3). Hauer, teaches that the same reaction that produces the colorimetric change responsible for the change in absorbance also produces corresponding hydrogen ions (p. 114, para. 2). Thus, the ordinary artisan would understand that it is an inherent property of the Marshall pNPP plus alkaline phosphatase detection system that the amount of change in pH would increase over time just as the amount of absorbance increases over time . 07-21-aia AIA Claim 11 is rejected under 35 U.S.C. 103 as being unpatentable over Ikebukuro 6 (Electrochemical Detection of Protein Using a Double Aptamer Sandwich, Analytical Letters , 37(14): 2901-2909, 2004) in view of Pachauri (Biologically sensitive field-effect transistors: from ISFETs to NanoFETs, Essays Biochem , 60(1): 80-90, 2016) and Juang (Proton-ELISA: Electrochemical immunoassay on a dual-gated ISFET array, Biosensors and Bioelectronics , 117, 175-182, 2018), as evidenced by Rocchitta 7 (Enzyme Biosensors for Biomedical Applications: Strategies for Safeguarding Analytical Performances in Biological Fluids, Sensors , 16(6): 780, 1-52, 2016) . Regarding independent claim 11 , Ikebukuro teaches … A conjugate solution comprising: conjugates each having a binding substance fused with a label, the conjugates configured to bind to a target substance and exhibit a detectable phenomenon of a change in concentration of an ion, the binding substance is a nucleic acid aptamer having an ability to bind to the target substance which is a protein, the label is an enzyme causing a detectable phenomenon of a change in concentration of the hydrogen ion (Fig. 1; p. 2902, para. 4; p. 2903, para. 2; p. 2903, para. 3; Fig. 1: the label is glucose dehydrogenase; the molecule that is generated is gluconolactone; Rocchitta, which is directed to a biosensor using an aptamer labeled with glucose dehydrogenase, teaches that this reaction also generates hydrogen ions (p. 4, para. 2; p. 7, para. 1)); the binding substance and the label being fused with each other through a chemical substituent, the chemical substituent being avidin (Fig. 1: aptamer 2-GDH complex binds to target protein thrombin and converts glucose to gluconolactone; p. 2903, para. 3); and the binding substance is configured to bind to the target substance to form a target-substance bound conjugate within a solution containing the conjugate and the target substance (Fig. 1; p. 2902, para. 4; p. 2903, para. 2); wherein the conjugate solution contains target-substance bound conjugates without any unbound conjugates and the label of the target-substance bound conjugate is activated in the conjugate solution causing the detectable phenomenon of the change in concentration of a hydrogen ion to occur due to generation (p. 2902, para. 4; p. 2903, para. 2; p. 2905, para. 1: unbound conjugates were removed with washing; Fig. 1: the label is glucose dehydrogenase; the molecule that is generated is gluconolactone; Rocchitta: p. 4, para. 2; p. 7, para. 1). Finally, regarding the limitation requiring that the conjugate solution is configured for measurement of the detectable phenomenon with an ion-sensitive field effect transistor (ISFET), as noted, Ikebukuro teaches a biosensor sandwich assay which generates hydrogen ions, and also teaches that the aptamers are attached to an electrochemical detection device (Fig. 1), although the device is not an ISFET. Juang teaches a biosensor sandwich assay that generates hydrogen ions (Fig. 1), and also teaches that ISFETs are capable of detecting changes in solutions due to fluctuations resulting from changes in ion concentration, e.g., hydrogen ion concentration and pH (Fig. 1). Pachauri teaches that ISFETs have certain advantages compared to other electrochemical detection techniques when used in biosensor assays, including miniaturization and high sensitivity, both of which are useful in point-of-care devices (p. 85, last para. to p. 86, first para.). Prior to the effective filing date of the instant invention, it would have been prima facie obvious to modify the Ikebukuro assay and the corresponding conjugate solution to incorporate the Juang ISFET and the corresponding conjugate solution that is configured for measurement by an ISFET. The ordinary artisan would have been motivated to do so to achieve the expected advantages of a conjugate solution which is capable of being used in a miniaturized POC device and which is capable of being used in a highly sensitive assay, as taught by Pachauri. The ordinary artisan would have had an expectation of success as ISFETs are recognized in the art as being useful for detecting conjugate solutions which generate hydrogen ion concentrations, and because the design and modification of nucleic acid assays is well-known in the art . 07-21-aia AIA Claim 11 is rejected under 35 U.S.C. 103 as being unpatentable over Ikebukuro 8 (Electrochemical Detection of Protein Using a Double Aptamer Sandwich, Analytical Letters , 37(14): 2901-2909, 2004) in view of Pachauri (Biologically sensitive field-effect transistors: from ISFETs to NanoFETs, Essays Biochem , 60(1): 80-90, 2016) and Juang (Proton-ELISA: Electrochemical immunoassay on a dual-gated ISFET array, Biosensors and Bioelectronics , 117, 175-182, 2018) and Ma (Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow, Nature Biotechnology , 34(5): 528-531, 2016), as evidenced by Rocchitta 9 (Enzyme Biosensors for Biomedical Applications: Strategies for Safeguarding Analytical Performances in Biological Fluids, Sensors , 16(6): 780, 1-52, 2016) . Regarding independent claim 11 , Ikebukuro teaches … A conjugate solution comprising: conjugates each having a binding substance fused with a label, the conjugates configured to bind to a target substance and exhibit a detectable phenomenon of a change in concentration of an ion, the binding substance is a nucleic acid aptamer having an ability to bind to the target substance which is a protein, the label is an enzyme causing a detectable phenomenon of a change in concentration of the hydrogen ion (Fig. 1; p. 2902, para. 4; p. 2903, para. 2; p. 2903, para. 3; Fig. 1: the label is glucose dehydrogenase; the molecule that is generated is gluconolactone; Rocchitta, which is directed to a biosensor using an aptamer labeled with glucose dehydrogenase, teaches that this reaction also generates hydrogen ions (p. 4, para. 2; p. 7, para. 1)); and the binding substance is configured to bind to the target substance to form a target-substance bound conjugate within a solution containing the conjugate and the target substance (Fig. 1; p. 2902, para. 4; p. 2903, para. 2); wherein the conjugate solution contains target-substance bound conjugates without any unbound conjugates and the label of the target-substance bound conjugate is activated in the conjugate solution causing the detectable phenomenon of the change in concentration of a hydrogen ion to occur due to generation (p. 2902, para. 4; p. 2903, para. 2; p. 2905, para. 1: unbound conjugates were removed with washing; Fig. 1: the label is glucose dehydrogenase; the molecule that is generated is gluconolactone; Rocchitta: p. 4, para. 2; p. 7, para. 1). Finally, regarding the limitation requiring that the conjugate solution is configured for measurement of the detectable phenomenon with an ion-sensitive field effect transistor (ISFET), as noted, Ikebukuro teaches a biosensor sandwich assay which generates hydrogen ions, and also teaches that the aptamers are attached to an electrochemical detection device (Fig. 1), although the device is not an ISFET. Juang teaches a biosensor sandwich assay that generates hydrogen ions (Fig. 1), and also teaches that ISFETs are capable of detecting changes in solutions due to fluctuations resulting from changes in ion concentration, e.g., hydrogen ion concentration and pH (Fig. 1). Pachauri teaches that ISFETs have certain advantages compared to other electrochemical detection techniques when used in biosensor assays, including miniaturization and high sensitivity, both of which are useful in point-of-care devices (p. 85, last para. to p. 86, first para.). Further, Ikebukuro teaches the binding substance and the label being fused with each other (Fig. 1; p. 2903, para. 3), while Ma teaches that the fusion is through a nucleic acid binding protein, specifically a CRISPR complex (abstract: system based on dCas9 combined with sgRNA; Fig. 1: sgRNAs are bound to labels; p. 528, left col., para. 2: discusses fusing sgRNAs to aptamers; thus, sgRNA can be used to fuse together aptamers and labels). Prior to the effective filing date of the instant invention, it would have been prima facie obvious to modify the Ikebukuro assay and the corresponding conjugate solution to incorporate the Juang ISFET and the corresponding conjugate solution that is configured for measurement by an ISFET. The ordinary artisan would have been motivated to do so to achieve the expected advantages of a conjugate solution which is capable of being used in a miniaturized POC device and which is capable of being used in a highly sensitive assay, as taught by Pachauri. The ordinary artisan would have had an expectation of success as ISFETs are recognized in the art as being useful for detecting conjugate solutions which generate hydrogen ion concentrations, and because the design and modification of nucleic acid assays is well-known in the art. It would have been further obvious to incorporate the Ma sgRNA linker into the Ikebukuro composition, discussed above. Ma teaches that sgRNA is useful for attaching labels to aptamers. The selection of a known material based on its suitability for its intended purpose supports a prima facie obviousness determination. MPEP 2144.07. Further, the ordinary artisan would have been motivated to optimize the component linkages through routine experimentation to customize the assay as needed. The ordinary artisan would have had an expectation of success as attaching various kinds of labeling moieties to nucleic acids is well-known in the art . Conclusion 12-151-07 AIA 07-97 12-51-07 Claim s 1, 4-6, 8-11, 14, 16-20 and 25-28 are being examined, and are rejected. No claims are allowed. Any inquiry concerning this communication or earlier communications from the examiner should be directed to CAROLYN GREENE whose telephone number is (571)272-3240. The examiner can normally be reached M-Th 7:30-5:30 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, Gary Benzion can be reached at 571-272-0782. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300. Information regarding the status of published or unpublished applications may be obtained from Patent Center. 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If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000. /CAROLYN L GREENE/Primary Examiner, Art Unit 1681 Application/Control Number: 17/721,558 Page 2 Art Unit: 1681 Application/Control Number: 17/721,558 Page 3 Art Unit: 1681 Application/Control Number: 17/721,558 Page 4 Art Unit: 1681 Application/Control Number: 17/721,558 Page 5 Art Unit: 1681 Application/Control Number: 17/721,558 Page 6 Art Unit: 1681 Application/Control Number: 17/721,558 Page 7 Art Unit: 1681 Application/Control Number: 17/721,558 Page 8 Art Unit: 1681 Application/Control Number: 17/721,558 Page 9 Art Unit: 1681 Application/Control Number: 17/721,558 Page 10 Art Unit: 1681 Application/Control Number: 17/721,558 Page 11 Art Unit: 1681 Application/Control Number: 17/721,558 Page 12 Art Unit: 1681 Application/Control Number: 17/721,558 Page 13 Art Unit: 1681 Application/Control Number: 17/721,558 Page 14 Art Unit: 1681 Application/Control Number: 17/721,558 Page 15 Art Unit: 1681 Application/Control Number: 17/721,558 Page 16 Art Unit: 1681 Application/Control Number: 17/721,558 Page 17 Art Unit: 1681 Application/Control Number: 17/721,558 Page 18 Art Unit: 1681 Application/Control Number: 17/721,558 Page 19 Art Unit: 1681 Application/Control Number: 17/721,558 Page 20 Art Unit: 1681 Application/Control Number: 17/721,558 Page 21 Art Unit: 1681 Application/Control Number: 17/721,558 Page 22 Art Unit: 1681 Application/Control Number: 17/721,558 Page 23 Art Unit: 1681 Application/Control Number: 17/721,558 Page 24 Art Unit: 1681 Application/Control Number: 17/721,558 Page 25 Art Unit: 1681 1 Sullenger was cited in the PTO-892 Notice of References Cited mailed November 3, 2025. 2 Samson was cited in the PTO-892 Notice of References Cited mailed March 27, 2025. 3 Ma was cited in the PTO-892 Notice of References Cited mailed November 3, 2025. 4 Samson was cited in the PTO-892 Notice of References Cited mailed March 27, 2025. 5 Lu was cited in the Information Disclosure Statement submitted September 23, 2022. 6 Ikebukuro was cited in the PTO-892 Notice of References Cited mailed March 27, 2025. 7 Rocchitta was cited in the PTO-892 Notice of References Cited mailed January 29, 2025. 8 Ikebukuro was cited in the PTO-892 Notice of References Cited mailed March 27, 2025. 9 Rocchitta was cited in the PTO-892 Notice of References Cited mailed January 29, 2025.
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