DETAILED ACTION
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 .
Claims 1, 3-6, 10-11, and 13-24 are pending. Claims 1 and 23 are amended. Claim 24 is new. Claims 2, 7-9, and 12 are canceled. Claims 1, 3-6, 10-11, and 13-24 are under examination.
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 12/18/25 has been entered.
Priority
The instant application 17/721,686 filed on 4/15/22 claims foreign priority to JP 2021-071057 filed on 4/20/21 and JP 2021-213139 filed on 12/27/21. The priority date is determined to be 4/15/22 in the absence of a copy certified translation of JP 2021-071057 and/or JP 2021-213139.
Receipt is acknowledged of JP 2021-071057 and JP 2021-213139 certified copies of papers required by 37 CFR 1.55. Priority Documents were electronically retrieved by USPTO from participating IP office on 6/16/22.
Should applicant desire to obtain the benefit of foreign priority under 35 U.S.C. 119(a)-(d) prior to declaration of an interference, a certified English translation of the foreign application must be submitted in reply to this action. 37 CFR 41.154(b) and 41.202(e).
Failure to provide a certified translation may result in no benefit being accorded for the non-English application.
Response to Arguments
Applicant’s arguments, see pages 9-10, filed 12/18/25, with respect to the rejections of claims 1, 3-6, 10-11, and 13-23 under 35 USC 103 have been fully considered but they are not persuasive. The 35 USC 103 rejections documented in the Final Rejection mailed on 11/3/25 have been revised to address claim amendments and new claim 24 filed 12/18/25 in this Non-Final Office Action. More detailed responses to Applicant’s arguments are provided at the end of each maintained rejection.
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.
Claims 1, 3-5, 10, 16-18, and 21-23 remain/are rejected under 35 U.S.C. 103 as being unpatentable over Wu et al. (2016; NPL citation U in PTO-892 filed 11/3/25; "Screening and development of DNA aptamers as capture probes for colorimetric detection of patulin"; Analytical Biochemistry 508 (2016) 58-64; http://dx.doi.org/10.1016/j.ab.2016.05.024) in view of Schmitz et al. (2020; NPL citation U in PTO-892, page 2, filed 5/14/25; "A SARS-CoV-2 spike binding DNA aptamer that inhibits pseudovirus infection in vitro by an RBD independent mechanism". bioRxiv preprint. https://doi.org/10.1101/2020.12.23.424171) and Juang et al. (2018; NPL citation 1 on IDS filed on 4/15/22; “Proton-ELISA: Electrochemical immunoassay on a dual-gated ISFET array”; Biosensors and Bioelectronics 117 (2018) 175–182. https://doi.org/10.1016/j.bios.2018.06.012).
This rejection is necessitated by claim amendments filed 12/18/25.
(i) Wu et al. teaches limitations relevant to claims 1, 3-5, 18, and 21-23.
Relevant to claim 1, Wu et al. Abstract teaches "Patulin (PAT) is a kind of mycotoxin that has serious harmful impacts on both food quality and human health. A high-affinity ssDNA aptamer that specifically binds to patulin was generated using systemic evolution of ligands by exponential enrichment (SELEX) assisted by graphene oxide (GO)… the sequence PAT-11 bound to patulin with high affinity and excellent selectivity…"
Further relevant to claim 1, Wu et al. teaches that "In first stage, the capture probes (aptamer and glucose oxidase immobilized on AuNPs) hybridized with the separation probes (cDNA immobilized on MNPs) and formed the sandwich complex. In the second stage, glucose oxidase-catalyzed oxidization of glucose led to the formation of gluconic acid and hydrogen peroxide (H2O2). The latter can catalytically oxidize iron (II) to iron (III), which can rapidly coordinate with squaric acid (SQA). Formation of the iron squarate complex causes the color of the solution to change to brownish red accompanying the increasing absorbance (l ¼ 469 nm) of the generated SQA-iron (III) chelate [citation]. In the presence of patulin, the aptamer preferentially bonds with the target and causes the partial capture of probes and the dissociation of separation probes. With an external magnetic field, the remaining capture probes were resuspended in buffer after separation. Accompanying the capture probes, the carried glucose oxidase can trigger the enzymatic catalytic reaction to produce the colored product. The change in the color/absorbance indirectly depends on the concentration of target patulin in the sample. By monitoring the shift in absorbance, we can quantitatively determine the concentration of target patulin in the sample" (page 62, column 1 continued to first paragraph of column 2).
The Wu et al. magnetic field reads on the instant removing a remainder of the plurality of conjugates that is not bound to the target substance in a state where the target substance is not immobilized on the solid phase.
Further relevant to claim 1, Wu et al. teaches "the capture probes were assembled by aptamer PAT-11 and glucose oxidase immobilized on gold nanoparticles (AuNPs)" (page 60, first paragraph of Section "Aptamer-based colorimetric bioassay"). The Wu et al. capture probes with an aptamer (binding substance) and enzyme (label) read on the instant conjugates. Notably, the label is conjugated with the binding substance via the AuNPs.
Further relevant to claim 1, Wu et al. Table 1 teaches that the PAT-11 DNA aptamer contains 40 nucleic acid bases.
The Wu et al. PAT-11 aptamer has an activity to bind to the target substance, a molecular weight less than a molecular weight of an immunoglobulin, and is a nucleic acid aptamer.
Further relevant to claim 1, Wu et al. teaches that the capture probes and patulin were incubated together in solution, without the patulin target substance immobilized on a solid phase (page 60, second paragraph of Section "Aptamer-based colorimetric bioassay"). As noted, the capture probes contain both the binding substance (aptamer) and label (glucose oxidase). Thus, binding the target substance enables binding the label with the target substance, as both components are brought into proximity via capture probe.
Further relevant to claim 1, Wu et al. Figure 1 teaches that the glucose acid catalyzes a detectable phenomenon of a change in ion concentrations when the target-aptamer-capture probes come into contact.
Thus, the Wu et al. teachings read on claim 1 A method for detecting a target substance using a plurality of conjugates, each conjugate including a binding substance and a label conjugated with the binding substance… the binding substance having an activity to bind to the target substance, having a molecular weight less than a molecular weight of an immunoglobulin, and being a low-molecular weight protein formulation or a nucleic acid aptamer… a number of bases of the nucleic acid aptamer falling within a range between 10, inclusive, and 100, inclusive, and the label causing a detectable phenomenon, which is a change in a concentration of an ion, the method comprising: binding the label of at least some of the plurality of conjugates with the target substance through the binding substance while not immobilizing the target substance on a solid phase; removing a remainder of the plurality of conjugates that is not bound to the target substance in a state where the target substance is not immobilized on the solid phase.
Relevant to claim 3, Wu et al. teaches "the capture probes were assembled by aptamer PAT-11 and glucose oxidase immobilized on gold nanoparticles (AuNPs)" (page 60, first paragraph of Section "Aptamer-based colorimetric bioassay").
This teaching reads on claim 3 wherein the label is at least one selected from the group consisting of an enzyme… and a nanoparticle.
Relevant to claims 4-5, Wu et al. Figure 1 teaches that the ion is a hydrogen ion and that the concentration of the ion changes due to generation… of the ion.
This teaching reads on claim 4 wherein the ion is at least one selected from the group consisting of hydrogen ion… and the concentration of the ion changes due to generation… of the ion; and claim 5 wherein the label is an enzyme that induces generation… of the ion.
Relevant to claim 18, Wu et al. Title teaches "Screening and development of DNA aptamers as capture probes for colorimetric detection of patulin". This teaching reads on claim 18 wherein the nucleic acid aptamer is one of a DNA aptamer.
Relevant to claim 21, Wu et al. teaches that the capture probes and patulin were incubated together in solution, allowing the plurality of conjugates and the target substance to coexist in a solution (page 60, second paragraph of Section "Aptamer-based colorimetric bioassay").
This teaching reads on claim 21 wherein binding at least some of the plurality of conjugates with the target substance includes allowing the plurality of conjugates and the target substance to coexist in a solution.
Relevant to claim 22, Wu et al. teaches that "In first stage, the capture probes (aptamer and glucose oxidase immobilized on AuNPs) hybridized with the separation probes (cDNA immobilized on MNPs) and formed the sandwich complex. In the second stage, glucose oxidase-catalyzed oxidization of glucose led to the formation of gluconic acid and hydrogen peroxide (H2O2). The latter can catalytically oxidize iron (II) to iron (III), which can rapidly coordinate with squaric acid (SQA). Formation of the iron squarate complex causes the color of the solution to change to brownish red accompanying the increasing absorbance (l ¼ 469 nm) of the generated SQA-iron (III) chelate [citation]. In the presence of patulin, the aptamer preferentially bonds with the target and causes the partial capture of probes and the dissociation of separation probes. With an external magnetic field, the remaining capture probes were resuspended in buffer after separation. Accompanying the capture probes, the carried glucose oxidase can trigger the enzymatic catalytic reaction to produce the colored product. The change in the color/absorbance indirectly depends on the concentration of target patulin in the sample. By monitoring the shift in absorbance, we can quantitatively determine the concentration of target patulin in the sample" (page 62, column 1 continued to first paragraph of column 2).
This Wu et al. disclosure teaches that the target substance is captured by the AuNPs - not the magnetic bead separation probes - before removing the remainder of the plurality of conjugates. This teaching reads on claim 22 wherein the target substance is not captured by a magnetic bead before removing the remainder of the plurality of conjugates.
Relevant to claim 23, Wu et al. Abstract teaches "Patulin (PAT) is a kind of mycotoxin that has serious harmful impacts on both food quality and human health. A high-affinity ssDNA aptamer that specifically binds to patulin was generated using systemic evolution of ligands by exponential enrichment (SELEX) assisted by graphene oxide (GO)… the sequence PAT-11 bound to patulin with high affinity and excellent selectivity…"
Further relevant to claim 23, Wu et al. teaches that "In first stage, the capture probes (aptamer and glucose oxidase immobilized on AuNPs) hybridized with the separation probes (cDNA immobilized on MNPs) and formed the sandwich complex. In the second stage, glucose oxidase-catalyzed oxidization of glucose led to the formation of gluconic acid and hydrogen peroxide (H2O2). The latter can catalytically oxidize iron (II) to iron (III), which can rapidly coordinate with squaric acid (SQA). Formation of the iron squarate complex causes the color of the solution to change to brownish red accompanying the increasing absorbance (l ¼ 469 nm) of the generated SQA-iron (III) chelate [citation]. In the presence of patulin, the aptamer preferentially bonds with the target and causes the partial capture of probes and the dissociation of separation probes. With an external magnetic field, the remaining capture probes were resuspended in buffer after separation. Accompanying the capture probes, the carried glucose oxidase can trigger the enzymatic catalytic reaction to produce the colored product. The change in the color/absorbance indirectly depends on the concentration of target patulin in the sample. By monitoring the shift in absorbance, we can quantitatively determine the concentration of target patulin in the sample" (page 62, column 1 continued to first paragraph of column 2).
The Wu et al. magnetic field reads on the instant removing a remainder of the plurality of conjugates that is not bound to the target substance in a state where the target substance is not immobilized on the solid phase.
Further relevant to claim 23, Wu et al. teaches "the capture probes were assembled by aptamer PAT-11 and glucose oxidase immobilized on gold nanoparticles (AuNPs)" (page 60, first paragraph of Section "Aptamer-based colorimetric bioassay"). The Wu et al. capture probes with an aptamer (binding substance) and enzyme (label) read on the instant conjugates. Notably, the label is conjugated with the binding substance via the AuNPs.
Further relevant to claim 23, Wu et al. Table 1 teaches that the PAT-11 DNA aptamer contains 40 nucleic acid bases.
The Wu et al. PAT-11 aptamer has an activity to bind to the target substance, a molecular weight less than a molecular weight of an immunoglobulin, and is a nucleic acid aptamer.
Further relevant to claim 23, Wu et al. teaches that the capture probes and patulin were incubated together in solution, without the patulin target substance immobilized on a solid phase (page 60, second paragraph of Section "Aptamer-based colorimetric bioassay"). As noted, the capture probes contain both the binding substance (aptamer) and label (glucose oxidase). Thus, binding the target substance enables binding the label with the target substance, as both components are brought into proximity via capture probe.
Further relevant to claim 23, Wu et al. Figure 1 teaches that the glucose acid catalyzes a detectable phenomenon of a change in hydrogen ion concentrations when the target-aptamer-capture probes come into contact.
Thus, the Wu et al. teachings read on claim 23 A method for detecting a target substance using a plurality of conjugates, each conjugate including a binding substance and a label conjugated with the binding substance… the binding substance having an activity to bind to the target substance, having a molecular weight less than a molecular weight of an immunoglobulin, and consisting of a nucleic acid aptamer, a number of bases of the nucleic acid aptamer falls within a range between 10, inclusive, and 100, inclusive, and the label causing a detectable phenomenon, which is a generation of a hydrogen ion, the method comprising: binding the label of at least some of the plurality of conjugates with the target substance through the binding substance while not immobilizing the target substance on a solid phase; removing a remainder of the plurality of conjugates that is not bound to the target substance in a state where the target substance is not immobilized on the solid phase.
(ii) Although Wu et al. is silent to specifics regarding the target substance being a viral envelope protein (claims 1 and 23), a low-molecular weight protein formulation (claims 1 and 16-17), or SARS-CoV-2 (claim 10) these limitations are known in the prior art and taught by Schmitz et al.
Schmitz et al. teaches "A SARS-CoV-2 spike binding DNA aptamer that inhibits pseudovirus infection in vitro by an RBD independent mechanism" (Title).
The receptor binding domain (RBD) of the spike glycoprotein of the coronavirus SARS-CoV-2 (CoV2-S) is a component viral envelope protein.
Relevant to claims 1, 10, 16-17, and 23, Schmitz et al. teaches "Having shown SP6 interacts with CoV2-S without interfering with complex formation with its cellular receptor ACE2, we next studied the impact of SP6 on viral infection. To address this question, we used the established VSV-ΔG*-based pseudotype system [citations] and generated Cov2-S and VSV-G pseudotyped virus particles. The interaction of SP6 with the CoV2-S pseudotyped virus was verified by an enzyme-linked oligonucleotide assay (ELONA) [citation]. In this experiment, the CoV2-S protein or the CoV2-S pseudotyped virus were captured by a nanobody binding to the RBD of CoV2-S and after washing the bound protein or pseudovirus particles were detected by adding biotinylated SP6, streptavidin-horse radish peroxidase (HRP) conjugates and its substrate 2,2′-Azino-di(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) (Supporting Fig. 4). We observed a concentration dependent increase in signal when SP6 and SP6.34 were used for detection, but not when employing SP6C and SP6.34C (Supporting Fig. 4a). Likewise, SP6 but not SPC6C detected the CoV2-S pseudotyped virus"
The Schmitz et al. aptamer SP6 detects the claims 1 and 23 target substance viral envelope protein without the envelope protein being separated from the virus; wherein the virus is SARS-CoV-2 (claim 10); and wherein a low-molecular weight protein formulation of a nanobody (claim 1 and 16), which is neither a monoclonal antibody nor a polyclonal antibody (claim 17).
(iii) Although Wu et al. and Schmitz et al. are silent to specifics regarding detection via ion-sensitive field effect transistor (claims 1 and 23), this limitation is known in the prior art and taught by Juang et al.
Relevant to claims 1 and 23, Juang et al. Abstract teaches “an electrochemical immunoassay platform called Proton-ELISA (H-ELISA) for the detection of bioanalytes. H-ELISA uniquely utilizes protons as an immunoassay detection medium, generated by the enzyme glucose oxidase… A proton-sensitive dual-gated ion-sensitive field effect transistor (DG-ISFET) sensor was also developed for sensitive and accurate detection of the proton signal in H-ELISA.”
This teaching reads on claim 1 detecting the target substance by an ion-sensitive field effect transistor detecting the detectable phenomenon; and claim 23 detecting the target substance by an ion-sensitive field effect transistor detecting the detectable phenomenon.
(iv) Although Wu et al. does not include the Schmitz et al. limitations (viral envelope protein target substance, low-molecular weight protein formulation, SARS-CoV-2) or the Juang et al. limitation (ion-sensitive field effect transistor), they would have been prima facie obvious to the skilled artisan. It is noted that Wu et al., Schmitz et al., and Juang et al. are analogous disclosures to the instant method for detecting target substances.
The skilled artisan would have been motivated to combine the analogous disclosures. Schmitz et al. Abstract teaches “The receptor binding domain (RBD) of the spike glycoprotein of the coronavirus SARS-CoV-2 (CoV2-S) binds to the human angiotensin converting enzyme 2 (ACE2) representing the initial contact point for leveraging the infection cascade. We used an automated selection process and identified an aptamer that specifically interacts with CoV2-S. The aptamer does not bind to the RBD of CoV2-S and does not block the interaction of CoV2-S with ACE2. Notwithstanding, infection studies revealed potent and specific inhibition of pseudoviral infection by the aptamer. The present study opens up new vistas in developing SARS-CoV2 infection inhibitors, independent of blocking the ACE2 interaction of the virus and harnesses aptamers as potential drug candidates and tools to disentangle hitherto inaccessible infection modalities, which is of particular interest in light of the increasing number of escape mutants that are currently being reported.” Schmitz et al. teaches “The coronavirus SARS-CoV-2 binds via its spike protein (CoV2-S) to the extracellular domain of the human angiotensin-converting enzyme 2 (ACE2) initiating the entry process into target cells” (page 1 of Introduction).
Thus, the skilled artisan would have been motivated to include the Schmitz et al. viral envelope protein target substance, low-molecular weight protein formulation, and SARS-CoV-2 limitations within the Wu et al. detection methodology because Schmitz et al. teaches that the viral envelope protein is critical to the virus’s entry into target cells, and would thus be critical in the viral pathogenesis and disease progression. Additionally, the skilled artisan would be motivated to examine SARS-CoV-2 within the Wu et al. detection methodology because Schmitz et al. teaches that escape mutants are increasingly being reported and that there is “particular interest” amongst other skilled artisans. The skilled artisan would have been further motivated to use the Schmitz et al. low-molecular weight nanobody within the Wu et al. methodology because Schmitz et al. Supporting Figure 4 depicts successful nanobody-detection of the SARS-CoV-2 target.
The skilled artisan would have been motivated to use the Juang et al. ion-sensitive field effect transistor within the Wu et al. methodology because Juang et al. teaches that it is capable of providing a separate, non-colorimetric measurement of the GOx-mediated ionic changes that the Wu et al. methodology generates. The skilled artisan would be further motivated by the Juang et al. Abstract teaching that “the platform is compatible with complex biological sample conditions such as human serum, suggesting that the platform is sufficiently robust for potential diagnostic applications.”
The skilled artisan would have a reasonable expectation of success based on the disclosures of Wu et al. in view of Schmitz et al. and Juang et al., as discussed in the preceding paragraphs.
Applicant’s Arguments
Applicant argues that “In amended claims 1, the binding substance 9 and the label 11 are conjugated, i.e. directly bonded to each other. This allows the size of the conjugate to be reduced, suppresses interference from other substances, and facilitates binding to the target substance 1. On the other hand, in Wu et al., the aptamer and glucose are bound to gold nanoparticles (AuNPs) to form a capture probe. In other words, the aptamer and the glucose oxidase are connected via AuNPs… Therefore Wu et al. does not disclose the binding substance and the label being conjugated” (Remarks 12/18/25, last paragraph of page 9 continued to first paragraph of page 10).
Response to Applicant’s Arguments
The rejection of claims 1, 3-5, 10, 16-18, and 21-23 under 35 USC 103 as being unpatentable over Wu et al. in view of Schmitz et al. and Juang et al. has been revised as stated above in this Non-Final Office Action that specifically addresses the amended limitations of claims 1 and 23 recited in the Amendments filed 12/18/25.
The Examiner respectfully disagrees with the assertion that “Wu et al. does not disclose the binding substance and the label being conjugated.” The broadest reasonable interpretation of the amended claims 1 and 23 limitations of “a label conjugated with the binding substance” includes any chemical linkage of the label and binding substance. The broadest reasonable interpretation of “conjugated with” includes both indirect and direct linkages. The Wu et al. label and binding substance are conjugated via the AuNPs, serving as a single, connected conjugate molecule. Thus, this amended limitation is not restricted to the argued – and not claimed – direct bond between label and binding substance, as would be accomplished via direct covalent bonding or chemical linkage between the label and binding substance.
Claims 6, 11, and 24 remain/are rejected under 35 U.S.C. 103 as being unpatentable over Wu et al. (2016; NPL citation U in PTO-892 filed 11/3/25; "Screening and development of DNA aptamers as capture probes for colorimetric detection of patulin"; Analytical Biochemistry 508 (2016) 58-64; http://dx.doi.org/10.1016/j.ab.2016.05.024) in view of Schmitz et al. (2020; NPL citation U in PTO-892, page 2, filed 5/14/25; "A SARS-CoV-2 spike binding DNA aptamer that inhibits pseudovirus infection in vitro by an RBD independent mechanism". bioRxiv preprint. https://doi.org/10.1101/2020.12.23.424171) and Juang et al. (2018; NPL citation 1 on IDS filed on 4/15/22; “Proton-ELISA: Electrochemical immunoassay on a dual-gated ISFET array”; Biosensors and Bioelectronics 117 (2018) 175–182. https://doi.org/10.1016/j.bios.2018.06.012), as applied to claims 1, 3-5, 10, 16-18, and 21-23 above, and further in view of Kamiyama et al. (2016; NPL citation U in PTO-892, page 1, filed 5/14/25; "Versatile protein tagging in cells with split fluorescent protein". Nat Commun 7, 11046. https://doi.org/10.1038/ncomms11046).
The teachings of Wu et al. in view of Schmitz et al. and Juang et al. are applied to instantly rejected claims 6, 11, and 24 as they were applied to claims 1, 3-5, 10, 16-18, and 21-23 as rendering obvious a method for detecting a target substance.
Wu et al. in view of Schmitz et al. and Juang et al. is silent to specifics regarding fusing the binding substance and label with a nucleic acid binding protein (claims 6, 11, and 24). However, these limitations were known in the prior art and taught by Kamiyama et al.
Kamiyama et al. Figure 5A and associated caption teaches dCas9::GFP11x7+GFP1-10::VP64 and sgCXCR4 activation of CXCR4 gene, wherein the CXCR4 fluorescence is measured in Figure 5B, and visualized within Figure 5C fluorescently labelled cells.
This teaching reads on claim 6 the binding substance and the label are fused with each other through… a nucleic acid binding protein… the nucleic acid binding protein is at least one selected from the group consisting of… CRISPR; claim 11 the binding substance and the label are fused with each other through a nucleic acid binding protein, and the nucleic acid binding protein is CRISPR; and claim 24 a nucleic acid binding protein of the binding substance is conjugated with… a nucleic acid binding protein of the label.
Although Wu et al. in view of Schmitz et al. and Juang et al. does not include a CRISPR protein fusion of label and binding substance, it would have been prima facie obvious to the skilled artisan. It is noted that Wu et al., Schmitz et al., Juang et al., and Kamiyama et al. are all analogous disclosures to the instant method for detecting a target substance.
The skilled artisan would be motivated to include the Kamiyama et al. CRISPR protein fusion because, as seen in Kamiyama et al. Figure 5B, the dCas9::GFP11x7+GFP1-10::VP64 probe was able to selectively generate a signal for the protein detection, whereas the dCas9::VP64 probe (lacking the full CRISPR-fused probe design) was unable to result in comparable signal detection.
The skilled artisan would have a reasonable expectation of success based on the disclosures of Wu et al. in view of Schmitz et al. and Juang et al., and further in view of Kamiyama et al., as discussed in the preceding paragraphs.
Applicant’s Arguments
Applicant argues that “none of Schmitz, Juang, Kamiyama, Odeh, Moser, and Song cure the deficiencies of Wu. As such, the prior art fails to disclose every feature of amended independent claims 1 and 23. New dependent claim 24 is patentable over the prior art for the reasons noted with respect to independent claim 1” (Remarks 12/18/25, page 10, paragraph 3).
Response to Applicant’s Arguments
The rejection of claims 6, 11, and 24 under 35 USC 103 as being unpatentable over Wu et al. in view of Schmitz et al. and Juang et al., and further in view of Kamiyama et al. has been revised as stated above in this Non-Final Office Action that specifically addresses the amended limitations of claims 1 and 23 recited in the Amendments filed 12/18/25.
As discussed in the above Response to Applicant’s Arguments relevant to claims 1, 3-5, 10, 16-18, and 21-23, Wu et al. in view of Schmitz et al. and Juang et al. renders obvious a method for detecting a target substance. Thus, the only independent claims are obvious, and as such, all of the claims are obvious.
Claims 13, 15, and 24 remain/are rejected under 35 U.S.C. 103 as being unpatentable over Wu et al. (2016; NPL citation U in PTO-892 filed 11/3/25; "Screening and development of DNA aptamers as capture probes for colorimetric detection of patulin"; Analytical Biochemistry 508 (2016) 58-64; http://dx.doi.org/10.1016/j.ab.2016.05.024) in view of Schmitz et al. (2020; NPL citation U in PTO-892, page 2, filed 5/14/25; "A SARS-CoV-2 spike binding DNA aptamer that inhibits pseudovirus infection in vitro by an RBD independent mechanism". bioRxiv preprint. https://doi.org/10.1101/2020.12.23.424171) and Juang et al. (2018; NPL citation 1 on IDS filed on 4/15/22; “Proton-ELISA: Electrochemical immunoassay on a dual-gated ISFET array”; Biosensors and Bioelectronics 117 (2018) 175–182. https://doi.org/10.1016/j.bios.2018.06.012), as applied to claims 1, 3-5, 10, 16-18, and 21-23 above, and further in view of Odeh et al. (2020; NPL citation X in PTO-892, page 1, filed 5/14/25; "Aptamers Chemistry: Chemical Modifications and Conjugation Strategies". Molecules 2020, 25, 3; doi:10.3390/molecules25010003).
The teachings of Wu et al. in view of Schmitz et al. and Juang et al. are applied to instantly rejected claims 13, 15, and 24 as they were applied to claims 1, 3-5, 10, 16-18, and 21-23 as rendering obvious a method for detecting a target substance.
Wu et al. in view of Schmitz et al. and Juang et al. is silent to specifics regarding fusing the binding substance and label together with a chemical substituent (claims 13, 15, and 24). However, these limitations were known in the prior art and taught by Odeh et al.
Odeh et al. teaches “Aptamers Chemistry: Chemical Modifications and Conjugation Strategies” (Title).
Relevant to claims 13, 15, and 24, Odeh teaches a “click reaction between azide- and alkyne-bearing nucleosides (Figure 13)” (page 15, Section “3.3.2. Triazole Modification); “Thiol maleimide coupling chemistry or Michael addition of a thiol to a maleimide” (page 27, Section “4.3. Thiol Maleiimide and Related Chemistry”); and “N-hydroxysuccinimide-activated QD [quantum dot] was covalently linked to… aptamer” (last sentence of page 25 continued to first sentence of page 26).
These teachings read on claim 13 the binding substance and the label are fused with each other through a chemical substituent, and the chemical substituent is at least one selected from the group consisting of… azide, alkyne… thiol, N-hydroxysuccinimide, maleimide; claim 15 the binding substance and the label are fused with each other through a chemical substituent, and the chemical substituent is at least one selected from the group consisting of… N-hydroxysuccinimide; and claim 24 a chemical substituent… of the binding substance is conjugated with a chemical substituent… of the label.
Although Wu et al., Schmitz et al. and Juang et al. do not include these chemical substituent fusions, it would have been prima facie obvious to the skilled artisan. Wu et al. and Odeh et al. are analogous disclosures to the instant aptamer-enabled detection methodology.
The skilled artisan would find it obvious – and be motivated – to use the chemical substituents disclosed by Odeh et al., as Odeh et al. teaches that “introducing chemical modifications into nucleic acid libraries increases the interaction capabilities of aptamers and thereby their target spectrum [citation]. Modified aptamers may show improved chemical diversity relative to aptamers composed entirely of natural DNA or RNA nucleotides and expand their applications in diagnostics, therapeutics, and nanotechnology” (page 2, paragraph 2).
The skilled artisan would have a reasonable expectation of success based on the disclosures of Wu et al. in view of Schmitz et al. and Juang et al., and further in view of Odeh et al., as discussed in the preceding paragraphs.
Applicant’s Arguments
Applicant argues that “none of Schmitz, Juang, Kamiyama, Odeh, Moser, and Song cure the deficiencies of Wu. As such, the prior art fails to disclose every feature of amended independent claims 1 and 23. New dependent claim 24 is patentable over the prior art for the reasons noted with respect to independent claim 1” (Remarks 12/18/25, page 10, paragraph 3).
Response to Applicant’s Arguments
The rejection of claims 13, 15, and 24 under 35 USC 103 as being unpatentable over Wu et al. in view of Schmitz et al. and Juang et al., and further in view of Odeh et al. has been revised as stated above in this Non-Final Office Action that specifically addresses the amended limitations of claims 1 and 23 recited in the Amendments filed 12/18/25.
As discussed in the above Response to Applicant’s Arguments relevant to claims 1, 3-5, 10, 16-18, and 21-23, Wu et al. in view of Schmitz et al. and Juang et al. renders obvious a method for detecting a target substance. Thus, the only independent claims are obvious, and as such, all of the claims are obvious.
Claim 14 remains/is rejected under 35 U.S.C. 103 as being unpatentable over Wu et al. (2016; NPL citation U in PTO-892 filed 11/3/25; "Screening and development of DNA aptamers as capture probes for colorimetric detection of patulin"; Analytical Biochemistry 508 (2016) 58-64; http://dx.doi.org/10.1016/j.ab.2016.05.024) in view of Schmitz et al. (2020; NPL citation U in PTO-892, page 2, filed 5/14/25; "A SARS-CoV-2 spike binding DNA aptamer that inhibits pseudovirus infection in vitro by an RBD independent mechanism". bioRxiv preprint. https://doi.org/10.1101/2020.12.23.424171) and Juang et al. (2018; NPL citation 1 on IDS filed on 4/15/22; “Proton-ELISA: Electrochemical immunoassay on a dual-gated ISFET array”; Biosensors and Bioelectronics 117 (2018) 175–182. https://doi.org/10.1016/j.bios.2018.06.012), as applied to claims 1, 3-5, 10, 16-18, and 21-23 above, and further in view of Moser et al. (2020; NPL citation W in PTO-892, page 1, filed 5/14/25; “Complementary Metal-Oxide-Semiconductor Potentiometric Field-Effect Transistor Array Platform Using Sensor Learning for Multi-ion Imaging". Anal. Chem. 2020, 92, 5276-5285. https://dx.doi.org/10.1021/acs.analchem.9b05836).
The teachings of Wu et al. in view of Schmitz et al. and Juang et al. are applied to instantly rejected claim 14 as they were applied to claims 1, 3-5, 10, 16-18, and 21-23 as rendering obvious a method for detecting a target substance.
Wu et al. in view of Schmitz et al. and Juang et al. is silent to specifics regarding different ions detected by the ion-sensitive field-effect transistors (claim 14). However, these limitations were known in the prior art and taught by Moser et al.
Relevant to claim 14, Moser et al. teaches “This work describes an array of 1024 ion-sensitive field-effect transistors (ISFETs) using sensor-learning techniques to perform multi-ion imaging for concurrent detection of potassium, sodium, calcium, and hydrogen… The platform is then trained by inducing a change in single-ion concentration and measuring the responses of all pixels.” (Abstract). This teaching reads on claim 14 the ion is at least one selected from the group consisting of potassium ion, sodium ion, calcium ion… and the concentration of the ion changes due to generation, consumption or absorption of the ion.
Although Wu et al. in view of Schmitz et al. and Juang et al. does not measure the potassium, sodium, and calcium ionic changes of Moser et al., it would have been prima facie obvious to the skilled artisan. It is noted that Wu et al., Schmitz et al. and Juang et al., and Moser et al. are all analogous disclosures to the instant detection of a target substance.
The ion-sensitive field-effect transistor of Moser et al. would be able to measure the hydrogen ions generated by the Wu et al. methodology. Additionally, the skilled artisan would be motivated to measure the additional Moser et al. ions within an expanded Wu et al. methodology because Moser et al. teaches that “The platform is designed to be versatile, as the CMOS [complementary metal-oxide-semiconductor] sensors can be covered with polymeric ion-selective membranes to provide selectivity toward electrolytes of interest, which promotes the versatility of the solution to several applications… The platform can be extended to a wider range of species and other applications using the same hardware, but treating the surface with different polymeric coatings, and integrated as part of an end-to-end microfluidic device” (Conclusion, page 5284).
The skilled artisan would have a reasonable expectation of success based on the disclosures of Wu et al. in view of Schmitz et al. and Juang et al., and further in view of Moser et al., as discussed in the preceding paragraphs.
Applicant’s Arguments
Applicant argues that “none of Schmitz, Juang, Kamiyama, Odeh, Moser, and Song cure the deficiencies of Wu. As such, the prior art fails to disclose every feature of amended independent claims 1 and 23. New dependent claim 24 is patentable over the prior art for the reasons noted with respect to independent claim 1” (Remarks 12/18/25, page 10, paragraph 3).
Response to Applicant’s Arguments
The rejection of claim 14 under 35 USC 103 as being unpatentable over Wu et al. in view of Schmitz et al. and Juang et al., and further in view of Moser et al. has been revised as stated above in this Non-Final Office Action that specifically addresses the amended limitations of claims 1 and 23 recited in the Amendments filed 12/18/25.
As discussed in the above Response to Applicant’s Arguments relevant to claims 1, 3-5, 10, 16-18, and 21-23, Wu et al. in view of Schmitz et al. and Juang et al. renders obvious a method for detecting a target substance. Thus, the only independent claims are obvious, and as such, all of the claims are obvious.
Claims 19-20 remain/are rejected under 35 U.S.C. 103 as being unpatentable over Wu et al. (2016; NPL citation U in PTO-892 filed 11/3/25; "Screening and development of DNA aptamers as capture probes for colorimetric detection of patulin"; Analytical Biochemistry 508 (2016) 58-64; http://dx.doi.org/10.1016/j.ab.2016.05.024) in view of Schmitz et al. (2020; NPL citation U in PTO-892, page 2, filed 5/14/25; "A SARS-CoV-2 spike binding DNA aptamer that inhibits pseudovirus infection in vitro by an RBD independent mechanism". bioRxiv preprint. https://doi.org/10.1101/2020.12.23.424171) and Juang et al. (2018; NPL citation 1 on IDS filed on 4/15/22; “Proton-ELISA: Electrochemical immunoassay on a dual-gated ISFET array”; Biosensors and Bioelectronics 117 (2018) 175–182. https://doi.org/10.1016/j.bios.2018.06.012), as applied to claims 1, 3-5, 10, 16-18, and 21-23 above, and further in view of Song et al. (2020; NPL citation 2 on IDS filed on 4/15/22; “Discovery of Aptamers Targeting the Receptor-Binding Domain of the SARS-CoV-2 Spike Glycoprotein”. Anal. Chem. 2020, 92, 9895−9900. DOI: 10.1021/acs.analchem.0c01394).
The teachings of Wu et al. in view of Schmitz et al. and Juang et al. are applied to instantly rejected claims 19-20 as they were applied to claims 1, 3-5, 10, 16-18, and 21-23 as rendering obvious a method for detecting a target substance.
Wu et al. in view of Schmitz et al. and Juang et al. is silent to specifics regarding chemically synthesizing the binding substance (claims 19-20). However, these limitations were known in the prior art and taught by Song et al.
Relevant to claims 19-20, Song et al. teaches that “The smaller size of aptamers (about 2−3 nm in diameter), as compared to antibodies (about 12−15 nm in diameter), subjects them to less steric hindrance on the surface of coronavirus (about 100 nm in diameter). In theory, the smaller size allows for the binding of more recognition molecules on the same surface area of coronavirus. Due to the chemical nature of nucleic acid, aptamers can be chemically synthesized, precisely modified, and high thermally stable and possess little batch-to-batch variation. These traits make for convenient transportation, storage, and standardization. Additionally, aptamers can be combined with other technologies to expand their performance and applications” (page 9895, column 2, paragraph 1).
This teaching reads on claim 19 chemically synthesizing the binding substance; and claim 20 chemically synthesizing the binding substance is an in vitro process.
Although Wu et al. methodology does not include chemically synthesizing the binding substance, it would have been prima facie to the skilled artisan to include the Song et al. binding substance limitations within the detection methodology rendered obvious by Wu et al. in view of Schmitz et al. and Juang et al. It is noted that Wu et al., Schmitz et al., Juang et al., and Song et al. are all analogous disclosures to the instant method for detection.
The skilled artisan would be motivated by the Song et al. teachings regarding the benefits of nucleic acid aptamer syntheses (overcoming steric hindrance, stability, standardization, extensive application; see page 9895, column 2, paragraph 1). Thus, the skilled artisan would be motivated to include the Song et al.-taught limitations within the detection methodology rendered obvious by Wu et al. in view of Schmitz et al. and Juang et al. to include chemical syntheses in order to take advantage of the “chemical nature of the nucleic acid”.
The skilled artisan would have a reasonable expectation of success based on the disclosures of Wu et al. in view of Schmitz et al. and Juang et al., and further in view of Song et al.
Applicant’s Arguments
Applicant argues that “none of Schmitz, Juang, Kamiyama, Odeh, Moser, and Song cure the deficiencies of Wu. As such, the prior art fails to disclose every feature of amended independent claims 1 and 23. New dependent claim 24 is patentable over the prior art for the reasons noted with respect to independent claim 1” (Remarks 12/18/25, page 10, paragraph 3).
Response to Applicant’s Arguments
The rejection of claim 19-20 under 35 USC 103 as being unpatentable over Wu et al. in view of Schmitz et al. and Juang et al., and further in view of Song et al. has been revised as stated above in this Non-Final Office Action that specifically addresses the amended limitations of claims 1 and 23 recited in the Amendments filed 12/18/25.
As discussed in the above Response to Applicant’s Arguments relevant to claims 1, 3-5, 10, 16-18, and 21-23, Wu et al. in view of Schmitz et al. and Juang et al. renders obvious a method for detecting a target substance. Thus, the only independent claims are obvious, and as such, all of the claims are obvious.
Conclusion
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/SARAH JANE KENNEDY/Examiner, Art Unit 1682
/WU CHENG W SHEN/Supervisory Patent Examiner, Art Unit 1682