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 .
Priority
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 5/16/2025 has been considered by the examiner.
Claim Objection
Claims 1, 7, 9, and 12-13 are objected to because of the following informalities:
Claim 1: please amend “the surface of a working electrode” to – [[the]] a surface of a working electrode of the one or more interdigitated planar working electrodes--.
Claim 7: please amend “each type of antibody” to -- each type of antibody of the one or more types of antibodies--.
Claim 9: please amend “ the solution of one or more antibodies” to -- the solution of the one or more antibodies--.
Claim 12: please amend “the antibody or antibodies” to -- the one or more types of antibodies--.
Claim 13: please amend “the protein is” to – the one or more SARS-COV-2 structural proteins [[is]] are --.
Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 1-14 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as failing to set forth the subject matter which the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the applicant regards as the invention.
Regarding claim 1, claim 1 recites “the ligands with anchored antibodies” and “the electrode”, which lack antecedent basis. It is unclear if the electrode refers to the working electrode or the one or more interdigitated planar working electrodes. Therefore, the scope of claim 1 is indefinite. Claims 2-14 are further rejected by virtue of their dependence upon and because they fail to cure the deficiencies of indefinite claim 1.
Regarding claim 5, claim 5 recites “the pathogen structural proteins”, which lacks antecedent basis. Therefore, the scope of claim 5 is indefinite. Claim 6 is further rejected by virtue of its dependence upon and because it fails to cure the deficiencies of indefinite claim 5.
Regarding claim 6, claim 6 recites “the protein”. It is unclear if it refers to “the pathogen structural proteins” or a specific protein of the pathogen structural proteins. Thus, the scope of claim 6 is indefinite.
Regarding claim 10, claim 10 recites “dissolved in a 0.5 M PBS or PBS/MES buffer solution”, and claim 1 recites “dissolved in MES buffer solution”, since PBS or PBS/MES buffer is different from the MES buffer, it is unclear what type of buffer solution is used. Furthermore, it is unclear if “(10 mg/ml)” is merely an example of the BSA concentration and therefore not required or is a required feature. Thus, the scope of claim 10 is indefinite. Claim 14 is further rejected by virtue of its dependence upon and because it fails to cure the deficiencies of indefinite claim 10.
Regarding claim 14, claim 14 recites “Use of an electrode of claim 10 for detecting pathogens in clinical or environmental samples”, which fails to recite active process steps to perform the use. Attempts to claim a process without setting forth any steps involved in the process generally raises an issue of indefiniteness under 35 U.S.C. 112(b) or pre-AIA 35 U.S.C. 112, second paragraph (see MPEP 2173.05(q). Thus, the scope of claim 14 is indefinite.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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 1-14 are rejected under 35 U.S.C. 103 as being unpatentable over Mojsoska et al. (Rapid SARS-CoV-2 detection using electrochemical immunosensor, Sensors, 2021, 21, 390), and in view of Hummer et al. (US20220011293A1), Torrente-Rodriguez et al. (SARS-CoV-2 rapidplex: a graphene-based multiplexed telemedicine platform for rapid and low-cost covid-19 diagnosis and monitoring, Matter, 2020, 3, 1981-1998) and Liyanage et al. (US20210140911A1). Mojsoska was provided in IDS filed on 5/16/2025.
Regarding claim 1, Mojsoska teaches a method for preparing a biomodified sensor for pathogen diagnostics (a method for preparing an electrochemical immunosensor for rapid SARS-CoV-2 detection [title, abstract, and section 2]), integrated in an electrochemical cell comprising a reference electrode (all electrochemical measurement were measured against a reference electrode [section 2.4]) and one or more interdigitated planar working electrodes (one planar graphene electrode as shown in Fig.1 [section 2.3]), comprising:
i) modifying the surface of a working electrode with a solution of an organic acid or a salt thereof (the graphene working electrode is functionalized with PBASE linker [Fig.1A and section 2.3]);
iii) depositing on the surface, a solution comprising one or more types of antibodies (The spike-specific antibodies are immobilized to the electrode [Fig.1B and section 2.3]);
iv) depositing on the surface containing the ligands with anchored antibodies, a concentrated solution of blocking agent dissolved in a buffer solution (BSA is used to block free surface or linkers on the electrode [Fig.1C]; the sensors were blocked by the addition of PBST containing 1% BSA to the working electrode [section 2.3]);
wherein the one or more types of antibodies are selected from among antibodies that bind to one or more types of structural proteins of the pathogen, one or more types of structural proteins genetically conserved among different variants or lineages of the pathogen, or combinations thereof (the spike-specific antibody binds to SARS-CoV-2 spike subunit 1 protein as shown in Fig.1 [sections 2.3 and 2.5]) and,
wherein the electrode is a graphene electrode (graphene working electrode in Fig.1).
Mojsoska further teaches the electrochemical measurements including EIS and CV were conducted to characterize the electrode and to evaluate the antibody-antigen interaction (section 2.4).
Mojsoska does not explicitly teach: (1) wherein the electrochemical cell further comprises a counter electrode; (2) the reference electrode, the counter electrode and the graphene working electrode are on a flexible substrate; (3) wherein the graphene working electrode is selected between an electrode printed on a flexible polymer film, by screen printing with carbon allotrope material ink or a graphene electrode induced by CO2 laser of a polyimide film; (4) ii) depositing on the modified surface a ligation solution comprising 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), a 2-morpholino ethane sulfonic acid buffer (MES) and N-hydroxysuccinimide (NHS) or N-hydroxysulfosuccinimide (sulfo-NHS); and (5) In the step iv, the buffer solution is MES buffer solution.
Hummer teaches methods and devices for detecting viruses and bacterial pathogens (title), wherein the viruses and bacterial pathogens include SARS-CoV-2 virus [para. 0094], and the electrochemical cell comprises a working electrode 1010, a reference electrode 1000, and a counter electrode 1020 on a substrate [para. 0120 ]. FIG. 16 shows the electrochemical detection device for COVID-19 SARS-CoV-2 virus in bodily fluids 1600. A nozzle deposits an electrically conductive electrode material 1604 on a Polyimide flexible substrate 1640 or other common thermoplastic polymers including Polyethylene terephthalate to make a printed sensor electrode 1610. A biologically sensitive molecule coating specific for SARS-CoV-2 is bound to the electrically conductive electrode 1620 to form a functionalized printed electrode. Coupled to the precision-printed sensor electrode 1610 are an external power source 1613 and an internal power source 1612. Also coupled to the printed sensor electrode 1610 is a solution compartment 1615 the receiver for the bodily fluid sample 1617 [para. 0134]. FIG. 44 shows a COVID-19 detection using a graphene sensor 4400 for an electrical current method 4401 of detecting selectable biologic analytical targets. The sensor structure can be of embodiments including a commercial CVD graphene on PET 4415 or print graphene ink on a polyimide substrate [para. 0205-0206]. Thus, Hummer teaches an electrochemical sensor for the detection of SARS-CoV-2, (1) wherein the electrochemical cell further comprises a counter electrode; (2) the reference electrode, the counter electrode and the graphene working electrode are on a flexible substrate; (3) wherein the graphene working electrode is an electrode printed on a flexible polymer film, by screen printing with carbon allotrope material ink (print graphene ink on a polyimide substrate).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the electrode cell of the electrochemical sensor in Mojsoska by: (1) providing a counter electrode; (2) having the reference electrode, the counter electrode and the graphene working electrode on a flexible polyimide substrate; (3) wherein the graphene working electrode is printed on the flexible polyimide film/substrate, as taught by Hummer, which teaches the suitable alternative electrochemical sensor for the detection of SARS-CoV-2 [para. 0205-0206; Fig.16 in Hummer].
Modified Mojsoska is silent to (4) ii) depositing on the modified surface a ligation solution comprising 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), a 2-morpholino ethane sulfonic acid buffer (MES) and N-hydroxysuccinimide (NHS) or N-hydroxysulfosuccinimide (sulfo-NHS); and (5) In the step iv, the buffer solution is MES buffer solution.
Torrente-Rodriguez teaches a graphene-based multiplexed telemedicine platform for rapid detection of SARS-CoV-2 (title and Fig.1), comprising four graphene WEs, one RE, and one CE on a PI substrate via CO2 laser engraving (section of Design of the SARS-CoV-2 RapidPlex Platform). Torrente-Rodriguez further teaches: i) modifying the surface of a working electrode with a solution of an organic acid or a salt thereof; ii) depositing on the modified surface a ligation solution comprising 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), a 2-morpholino ethane sulfonic acid buffer (MES) and N-hydroxysuccinimide (NHS) or N-hydroxysulfosuccinimide (sulfo-NHS) (A 10-mL aliquot of 5.0 mM PBA in DMF was drop-cast on the graphene surface and
incubated for 2 h at room temperature in a humid chamber. After rinsing with DMF,
IPA, and deionized (DI) water, and drying under air flow, electrodes were incubated
with 10 mL of a mixture solution containing 0.4 M EDC and 0.1 M sulfo-NHS in 0.025 M MES (pH 6.5) for 35 min at room temperature under humid ambient conditions [Functionalization of the SARS-CoV-2 RapidPlex and Electrochemical Readout]).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to substitute the PBASE in the step i) with PBA, and further providing the step ii) of depositing on the modified surface a ligation solution comprising 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysulfosuccinimide (sulfo-NHS), as taught by Torrente-Rodriguez, which teaches the suitable alternative method to functionalize the graphene surface for immobilizing the antibody onto the graphene surface for the detection of SARS-CoV-2 [Functionalization of the SARS-CoV-2 RapidPlex and Electrochemical Readout].
Modified Mojsoska is silent to: (5) In the step iv, the buffer solution is MES buffer solution.
Liyanage teaches a biosensor for detecting an analyte in a fluid sample (abstract), and further teaches a blocking agent of 2% BSA in MES buffer (10 mM, pH 6) [para. 0058].
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to substitute the PBS buffer in step iv with MES buffer such that the blocking agent of BSA is dissolved in MES buffer, as taught by Liyanage, which teaches the suitable alternative buffer for the blocking agent in a biosensor.
Regarding claim 2, Modified Mojsoska teaches the method of claim 1, wherein the organic acid or salt thereof is selected from pyrene butyric acid (PBA) (as outlined in the rejection of claim 1 above, the organic acid or salt is PBA [Fig.2 in Torrente-Rodriguez]).
Regarding claim 3, Modified Mojsoska teaches the method of claim 1, and Mojsoska teaches wherein the pathogen is a virus (SAR-CoV-2 virus [abstract]).
Regarding claim 4, Modified Mojsoska teaches the method of claim 3, and Mojsoska teaches wherein the virus is severe acute respiratory syndrome-causing coronavirus type 2 (SARS-COV-2) (SAR-CoV-2 virus [abstract]).
Regarding claim 5, Modified Mojsoska teaches the method of claim 4, and Mojsoska teaches wherein the pathogen structural proteins, are selected from the group comprising: spike(S), nucleocapsid (N), membrane (M) or envelope (E) protein of SARS-COV-2 virus or mixtures thereof (SARS-CoV-2 Spike S1-His protein [section 2.5]).
Regarding claim 6, Modified Mojsoska teaches the method of claim 5, and Mojsoska teaches wherein the protein is S protein, N protein or a mixture thereof (SARS-CoV-2 Spike S1-His protein [section 2.5]).
Regarding claim 7, Modified Mojsoska teaches the method of claim 1, and Mojsoska teaches wherein the anti-spike antibody was diluted 100x in PBS to approximately 22 μg/ml and was added to the working electrode (section 2.3), thus is silent to wherein each type of antibody is added at a concentration of 8 to 15 μg/ml.
However, generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature is critical. “[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." MPEP § 2144.05(II)(A). Therefore, it would have been obvious to one skilled in the art to use a concentration of 8 to 15 μg/ml of the antibody immobilized onto the electrode surface for the detection of SARS-CoV-2 spike.
Furthermore, Mojsoska teaches additional characterization of the actual number of antibodies bound to the surface many shed light on whether a denser antibody immobilization can lead to higher sensitivity (the last paragraph in section 3.4). Since a denser antibody requires a higher concentration of the antibody immobilized onto the electrode surface, thus the concentration of the antibody may affect the sensitivity.
As the detection sensitivity is a variable that may be modified, among others, by adjusting the concentration of the antibody immobilized on the electrode surface, the precise concentration of the antibody immobilized onto the electrode surface would have been considered a result effective variable by one having ordinary skill in the art before the effective filing date of the invention. As such, without showing unexpected results, the claimed concentration of the antibody immobilized onto the electrode surface being in a range from 8 to 15 μg/ml cannot be considered critical. Accordingly, one of ordinary skill in the art before the effective filing date of the invention would have optimized, by routine experimentation, the concentration of the antibody to obtain the desired concentration of the antibody in order to achieve the desired detection sensitivity. “[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation.” See In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). The discovery of an optimum value of a known result effective variable, without producing any new or unexpected results, is within the ambit of a person of ordinary skill in the art. See In re Boesch, 205 USPQ 215 (CCPA 1980) (see MPEP § 2144.05, II.).
Regarding claim 8, Modified Mojsoska teaches the method of claim 1, and Torrente-Rodriguez wherein in step ii) the ligation solution is incubated at a constant temperature between 4 and 25° C (electrodes were incubated for 35 min at room temperature [Functionalization of the SARS-CoV-2 RapidPlex and Electrochemical Readout]. Room temperature is generally ranged from 20 to 22 oC, thus the disclosed temperature falling within the claimed temperature).
Torrente-Rodriguez teaches wherein the incubation during the step ii) lasts for 35 min instead of 1 to 2 h.
During the step ii), since the EDC/sulfo-NHS activates the carboxylic groups on PBA to form the linker as shown in Fig.2A of Torrente-Rodriguez [Functionalization of the SARS-CoV-2 RapidPlex and Electrochemical Readout in Torrente-Rodriguez], the incubation time will affect the amount of carboxylic groups on PBA been activated, and accordingly the density of the formed linker which will further affect the amount of antibodies coupled to the linker during the step iii. Thus, one of ordinary skill in the art would recognize that the incubation time would affect the linker formation and accordingly its antibody binding capacities.
As the linker formation and antibody binding capacities are variables that can be modified, among others, by adjusting the incubation time of the step ii), the precise incubation time of step ii) would have been considered a result effective variable by one having ordinary skill in the art before the effective filing date of the invention. As such, without showing unexpected results, the claimed incubation time for 1 to 2h cannot be considered critical. Accordingly, one of ordinary skill in the art before the effective filing date of the invention would have optimized, by routine experimentation, the incubation time in order to achieve the desired linker density and antibody binding capacities. “[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation.” See In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). The discovery of an optimum value of a known result effective variable, without producing any new or unexpected results, is within the ambit of a person of ordinary skill in the art. See In re Boesch, 205 USPQ 215 (CCPA 1980) (see MPEP § 2144.05, II.).
Regarding claim 9, Modified Mojsoska teaches the method of claim 1, and Torrente-Rodriguez teaches wherein in step iii) the solution of one or more antibodies is incubated at a constant temperature between 4 and 25° C for 3h (specific antibodies were covalently attached onto activated surface and incubated for 3h at room temperature [Functionalization of the SARS-CoV-2 RapidPlex and Electrochemical Readout]).
Torrente-Rodriguez teaches wherein the incubation lasts for 3h during the step iii instead of 1 to 2 h.
Mojsoska teaches wherein the anti-spike antibody is incubated for 2h (section 2.3).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the incubation time from 3h to 2h, as taught by Mojsoska, which teaches the suitable alternative incubation time of the anti-spike antibody (section 2.3).
Regarding claim 10, Modified Mojsoska teaches the method of claim 1, and wherein in step iv) 1% concentrated solution of bovine serum albumin (BSA) dissolved in a 10 mM PBS or PBS/MES buffer solution at pH 6.0 is deposited and incubated at a constant temperature between 4 and 25° C. for 1 to 2 h (Mojsoska teaches wherein the sensors were blocked with 1% BSA in a buffer and incubated for 1h at room temperature [section 2.3]; as outlined in the rejection of claim 1 above, the buffer is 10 mM MES buffer at pH 6 [para.0058 in Liyanage]).
Modified Mojsoska is silent to wherein the concentration of the buffer solution is 0.5 M, and the BSA concentration is 10 mg/mL. However, generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature is critical. “[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." MPEP § 2144.05(II)(A). Therefore, it would have been obvious to one skilled in the art to use 10 mg/mL BSA dissolved in a 0.5 M MES buffer solution to make the solution for the blocking step.
Modified Mojsoska is silent to the volume of “4 μl” of 1% (10 mg/ml) concentrated solution of bovine serum albumin (BSA) dissolved in the MES buffer.
However, the volume of 4 μl of 1% (10 mg/ml) concentrated BSA dissolved in 0.5 M MES buffer further limits the final concentration of BSA in the solution for the blocking step. Generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature is critical. “[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." MPEP § 2144.05(II)(A).
Regarding claim 11, Modified Mojsoska teaches a biomodified electrode for pathogen detection prepared by the method of claim 1 (the biomodified graphene working electrode which is functionalized by PBA, EDC/Sulfo-NHS for detecting SARS-CoV-2, as outlined in the rejection of claim 1 above).
Regarding claim 12, Modified Mojsoska teaches the electrode of claim 10 (the graphene working electrode which is functionalized by PBA, EDC/Sulfo-NHS for detecting SARS-CoV-2, as outlined in the rejection of claim 10 above), wherein the antibody or antibodies selectively bind to one or more SARS-COV-2 structural proteins (the antibody selectively binds to SARS-CoV-2 Spike S1-His protein [section 2.5 in Mojsoska]).
Regarding claim 13, Modified Mojsoska teaches the electrode of claim 12, and Mojsoska teaches wherein the protein is selected from the group comprising SARS-COV-2 S protein and SARS-COV-2 N protein, or mixtures thereof (SARS-CoV-2 Spike S1-His protein [section 2.5]).
Regarding claim 14, Modified Mojsoska teaches use of an electrode of claim 10 for detecting pathogens in clinical or environmental samples (detection of SARS-CoV-2 Spike S1-His protein [sections 2.5 and 3.3-3.4 in Mojsoska]; Torrente-Rodriguez teaches successfully evaluated the applicability of the SARS-CoV-2 RapidPlex platform with COVID 19-positive and -negative blood and saliva samples [abstract]).
Conclusion
The prior arts made of record and not relied upon are considered pertinent to applicant's disclosure: Wibowo et al. (The electrical property of SiO2/graphene/PBA-NHS/anti O and K E. coli antibodies as sensing layer for Escherichia coli bacteria sensor, IOP Conf. Series: Materials Science and Engineering, 2020, 982, 012024) teaches PBA-NHS as a linker between antibody and graphene. Klimov et al. (US5770458A) teaches a blocking step of using blocking agent of BSA in MES for 1h at room temperature.
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/SHIZHI QIAN/Primary Examiner, Art Unit 1795
