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 .
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 6/17/2024 has been considered by the examiner.
Election/Restrictions
Applicant's election of Species B, claims 1-9 and 12-17, with traverse in the reply filed on 04/07/2026 is acknowledged.
The traversal is on the ground(s) that “the invention of Species A is basically the same as that of the other species, and any prior art searched for one group is applicable to the other group”. This is found persuasive; thus the previous restriction is withdrawn and claims 1-17 are examined in the current office action.
Claim Objection
Claims 1-2 and 11 are objected to because of the following informalities:
Claim 1: please amend “applying a sample” to -- applying [[a]] the sample--; “a chromogenic substrate the surface of the sensor” to -- a chromogenic substrate to the surface of the sensor--.
Claim 2: please amend “an electrode analysis device” to – [[an]] the electrode analysis device--.
Claim 11: please amend “step d)” to – the step d)--.
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 1-17 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 “wherein a possible working electrode-counter electrode current change is thus generated”. Even though the target biomarker is possibly present in the sample, the rest of the claim recites further limitations assuming it is present in the sample. It is unclear if a working electrode-counter electrode current change is possibly generated in the presence of the target biomarker. Thus, the scope of claim 1 is indefinite. Claims 2-17 are further rejected by virtue of their dependence upon and because they fail to cure the deficiencies of indefinite claim 1.
Regarding claim 17, claim 17 recites “further comprising vibrating the sample with a vibration exciter to excite at least one of the capture molecule, the plurality of blocking molecules, the sample, the target biomarker, the detection molecule, the detection molecule-target biomarker complex, the sandwich complex, the redox-capable enzyme, or the chromogenic substrate”. It is unclear if the first “the sample” refers to the sensor, otherwise it is unclear how to vibrate the sample by exciting other components such as the capture molecule, the plurality of blocking molecules, the target biomarker, the detection molecule, the detection molecule-target biomarker complex, the sandwich complex, the redox-capable enzyme, or the chromogenic substrate, that are different from the sample. Thus, the scope of claim 17 is indefinite.
Claim Rejections - 35 USC § 102
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 the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1-3, 5-14, and 16 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Sokolov et al. (US20210332405A1).
Regarding claim 1, Sokolov teaches a method of analyzing a sample using an electrochemical based sensor system (a method for detecting a binding event between an immobilised primary antibody and a target analyte-secondary antibody-enzyme complex using an electrochemical based sensor system as shown in Fig.2 [abstract; para. 0049; Fig.2]), the method comprising:
a) applying a sample possibly containing a target biomarker to a surface of an electrochemical based sensor (applying a sample containing CRP to a surface of an electrochemical based sensor as shown in Fig.2 [claim 19; para. 0035]; A secondary antibody that has been selected against another epitope of CRP and conjugated with an enzyme tag [e.g. HRP and ALP] is simultaneously incubated over the electrode array along with the sample [para. 0085]), wherein the electrochemical based sensor comprises:
i. an electrode analysis device (PalmSens 4 potentiostat [para. 0097]);
ii. a working electrode comprising a surface (an Au working electrode comprising a surface as shown in Fig.2 [para. 0013, 0090]), wherein capture molecules (capture antibody in Fig.2; an primary antibody immobilised on an electrode surface [para. 0013]; An anti-CRP antibody is physically adsorbed or chemically conjugated to the surface of the electrode [para. 0085]) are adsorbed onto the surface of the working electrode to generate an occupied capture surface (see Fig.2; An anti-CRP antibody is physically adsorbed or chemically conjugated to the surface of the electrode [para. 0085]) and a plurality of blocking molecules are adsorbed onto a remaining unoccupied capture surface (the primary antibody is bound to a surface and the rest of the surface deactivated using blocking proteins [para. 0046]);
iii. a counter electrode (a platinum wire counter electrode [para. 0097]);
iv. a pseudo-reference electrode (Ag/AgCl reference electrode [para. 0097]), wherein the working electrode, the counter electrode, the pseudo-reference electrode, and the electrode analysis device are electrically connected (Electrochemical measurements were performed using an Ag/AgCl reference electrode and a platinum wire counter electrode using PalmSens 4 potentiostat [para. 0097]; The electroactive precipitate may also be measured potentiometrically, either at open circuit or with application of a fixed current [chronopotentiometrically].These measurements are possible where the precipitate causes a change to the potential on the working electrode [para. 0090]; thus the working, counter and reference electrodes are electrically connected to the potentiostat for the electrochemical measurements);
b) applying a detection molecule (biotinylated secondary antibody in Fig.2) and a redox-capable enzyme (streptavidin modified polymeric HRP in Fig.2) to the surface of the sensor (contacting an primary antibody immobilized on an electrode surface with a target analyte and a secondary antibody and enzyme label in one or more solutions to create a sandwich complex bound to the electrode surface [para. 0013]; A secondary antibody that has been selected against another epitope of CRP and conjugated with an enzyme tag [e.g. HRP and ALP] is simultaneously incubated over the electrode array [para. 0085]), wherein the detection molecule is designed to bind to the target biomarker (Fig.2 shows the target biomarker CRP binds to the secondary antibody; the secondary antibody binds directly to the target analyte [para. 0025]; A secondary antibody that has been selected against another epitope of CRP [para. 0085]) to form a detection molecule-target biomarker complex (a CRP-secondary antibody complex is shown in Fig.2);
c) incubating the sample, the detection molecule and the redox-capable enzyme on the sensor for a period of time (A secondary antibody that has been selected against another epitope of CRP and conjugated with an enzyme tag [e.g. HRP and ALP] is simultaneously incubated over the electrode array along with the sample, that can be either serum, plasma, urine, saliva etc. for a duration of 5 minutes [para. 0085]); and “such that the target biomarker possibly present in the sample may bind the detection molecule to form the detection molecule-target biomarker complex” is an intended result of a positively recited step. The court noted that a "‘whereby clause in a method claim is not given weight when it simply expresses the intended result of a process step positively recited.’" Id. (quoting Minton v. Nat’l Ass’n of Securities Dealers, Inc., 336 F.3d 1373, 1381, 67 USPQ2d 1614, 1620 (Fed. Cir. 2003)); Sokolov further teaches the incubation forms the detection molecule-target biomarker complex as shown in Fig.2;
wherein the capture molecule and the detection molecule-target biomarker complex form a sandwich complex (contacting a primary antibody immobilized on an electrode surface with the single solution of the target analyte, the secondary antibody and the enzyme label to create a sandwich complex bound to the electrode surface [para. 0035]; a sandwich complex of capture antibody-CRP-secondary antibody is shown in Fig.2) upon binding of the capture molecule to the detection molecule-target biomarker complex, said sandwich complex comprising the capture molecule bound to the detection molecule-target biomarker complex (capture antibody bound to CRP-secondary antibody complex to form the sandwich complex as shown in Fig.2); and
d) applying a substrate solution comprising a chromogenic substrate the surface of the sensor (contacting the sandwich complex with a substrate for the HRP [para. 0035]; chromogenic substrates for HRP and AP are shown in Table 1 [para. 0052]);
wherein the redox-capable enzyme catalyzes a redox reaction in which the chromogenic substrate donates electrons to catalyze hydrogen peroxide into water (Fig.2 shows HRP catalyzes a redox reaction in which it breaks down H2O2 into water and DAB is oxidized by donating electrons to form an insoluble precipitate; the polymerisation of 3, 3′-Diaminobenzidine (DAB) [see FIG. 1] occurs via oxidation using hydrogen peroxide catalyzed by the horseradish peroxidase [HRP] enzyme [para. 0049]);
wherein the chromogenic substrate is thus oxidized to form an oxidized form of the chromogenic substrate (the polymerisation of 3, 3′-Diaminobenzidine [DAB] [see FIG. 1] occurs via oxidation using hydrogen peroxide catalyzed by the horseradish peroxidase enzyme [para. 0049]; the insoluble precipitate is deemed as the oxidized form of the DAB);
wherein the oxidized form of the chromogenic substrate comprises an electrically-detectable compound (the amount of insoluble precipitate on the surface [e.g. DAB] is determined by performing direct oxidation of the precipitate [e.g. DAB] in the solution by using cyclic voltammetry and properties of this trace are used to determine the signal [para. 0068]; The quantity of the precipitate [which is proportional to the amount of HRP-tagged secondary and thus proportional to the amount of antigen] can be measured electrochemically using standard faradaic impedance spectroscopy [para. 0085]; thus the insoluble precipitate comprises an electrically-detectable compound),
wherein the electrically-detectable compound is reduced to an unoxidized form of the chromogenic substrate via a working electrode reducing potential applied to the working electrode (The electroactive precipitate may be measured chronoamperometrically by holding a potential that causes oxidation or reduction of the precipitate and measuring the current through time [para. 0089]), said working electrode reducing potential transfers electrons from the working electrode to the electrically-detectable compound (holding a potential that causes reduction of the precipitate [para. 0089], thus the reducing potential transfer electrons from the working electrode to the precipitate), thus reducing the electrically-detectable compound to the unoxidized form of the chromogenic substrate (this is an intended result of a positively recited step. The court noted that a "‘whereby clause in a method claim is not given weight when it simply expresses the intended result of a process step positively recited.’" Id. (quoting Minton v. Nat’l Ass’n of Securities Dealers, Inc., 336 F.3d 1373, 1381, 67 USPQ2d 1614, 1620 (Fed. Cir. 2003)),
wherein a possible working electrode-counter electrode current change is thus generated (The electroactive precipitate may be measured chronoamperometrically by holding a potential that causes oxidation or reduction of the precipitate and measuring the current through time [para. 0089]; thus it generates a current change between the working and counter electrodes),
wherein said working electrode-counter electrode current change is measured as a function of time by a potentiostat using chronoamperometry (Electrochemical measurements were performed using PalmSens 4 potentiostat [para. 0097]; measuring the current through time [para. 0089]), and
wherein said working electrode-counter electrode current change may be used to perform at least one of detection of the target biomarker or quantification of the target biomarker (The quantity of the precipitate [which is proportional to the amount of HRP-tagged secondary and thus proportional to the amount of antigen] can be measured electrochemically using standard faradaic impedance spectroscopy [para. 0085]).
Regarding claim 2, Sokolov teaches the method of claim 1, wherein an electrode analysis device comprises a potentiostat (PalmSens 4 potentiostat [para. 0097]).
Regarding claim 3, Sokolov teaches the method of claim 1, wherein the working electrode is a gold electrode comprising a gold surface (Au electrode comprising a gold surface as shown in Fig.2; the electrode surface comprises gold, for example the surface is a gold surface [para. 0060]).
Regarding claim 5, Sokolov teaches the method of claim 1, wherein the pseudo-reference electrode is a Silver/Silver Chloride (Ag/AgCl) electrode ( an Ag/AgCl reference electrode [para. 0097]).
Regarding claim 6, Sokolov teaches the method of claim 1, wherein the capture molecule is a capture antibody (capture antibody in Fig.2).
Regarding claim 7, Sokolov teaches the method of claim 1, wherein the plurality of blocking molecules comprise bovine serum albumin molecules (Suitable blocking agents may include a protein such as bovine serum albumin [para. 0061]).
Regarding claim 8, Sokolov teaches the method of claim 1, wherein the detection molecule is a detection antibody (secondary antibody as shown in Fig.2).
Regarding claim 9, Sokolov teaches the method of claim 1, wherein the redox-capable enzyme is bound to the detection molecule (Fig.2 shows the HRP is bound to the secondary antibody).
Regarding claim 10, Sokolov teaches the method of claim 1, wherein the detection molecule is biotinylated (biotinylated secondary antibody as shown in Fig.2) and the redox-capable enzyme is bound to a probe comprising a streptavidin or avidin tetramer and forms a probe-enzyme conjugate (Suitable polymeric labels may include streptavidin-poly-HRP conjugates [para. 0055; Fig.2]).
Regarding claim 11, Sokolov teaches the method of claim 10, wherein the probe-enzyme conjugate is applied to the surface of the sensor before step d) (A secondary antibody that has been selected against another epitope of CRP and conjugated with an enzyme tag [e.g. HRP and ALP] is simultaneously incubated over the electrode array along with the sample, that can be either serum, plasma, urine, saliva etc. for a duration of 5 minutes. After the incubation step, a wash step is introduced to remove any unbound excess material (antibody or antigens). Precipitating solution [e.g. precipitating TMB or other chromogen precipitating materials], which create insoluble precipitates are added to the sensor for a controlled period of time and react with the enzyme to produce an electroactive precipitate on the electrode surface [para. 0087]; thus, the probe-enzyme conjugate is applied to the surface of the sensor to form the sandwich complex before the step d since applying the substrate solution in step d is after the incubation step), wherein the streptavidin or avidin tetramer of the probe-enzyme conjugate interacts with the biotinylated detection molecule of the sandwich complex (Fig.2 shows the streptavidin modified polymeric HRP bounds to the biotinylated secondary antibody of the sandwich complex).
Regarding claim 12, Sokolov teaches the method of claim 1, wherein the detection molecule is applied to the surface of the sensor before the sample is applied to the surface of the sensor (contacting a primary antibody immobilised on an electrode surface with the single solution of the target analyte, the secondary antibody and the enzyme label to create a sandwich complex bound to the electrode surface [para. 0035]; thus a primary antibody [detection molecule] is applied to the surface of the sensor before contacting the solution including the sample).
Regarding claim 13, Sokolov teaches the method of claim 1, wherein the redox-capable enzyme is horseradish peroxidase (HRP as shown in Fig.2; the horseradish peroxidase (HRP) enzyme [para. 0049]).
Regarding claim 14, Sokolov teaches the method of claim 1, wherein the chromogenic substrate comprises 3,3′,5,5′-tetramethylbenzidine (TMB), or 3,3′-Diaminobenzidine (DAB) (Any suitable substrate may be used, for example DAB [3, 3′-diaminobenzidine] or TMB [3,3′,5,5′-tetramethylbenzidine]; The substrate may be stabilised DAB, for example ImmPact™ DAB [para. 0020]).
Regarding claim 16, Sokolov teaches the method of claim 1, wherein the sample is a urine sample, a blood sample, or a saliva sample (the sample can be either serum, plasma, urine, saliva etc [para. 0085]).
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.
Claim 4 is rejected under 35 U.S.C. 103 as being unpatentable over Sokolov, as applied to claim 3 above, and further in view of Kim et al. (Gold-film array-electrode for electrochemical ELISA, Sensors and Actuators B, 2005, 111-112, 463-469).
Regarding claim 4, Sokolov teaches the method of claim 3, and further teaches wherein the counter electrode is a Pt electrode (a platinum wire counter electrode [para. 0097]), thus is silent to wherein the counter electrode is a gold electrode.
Kim teaches an electrochemical ELISA system comprising gold-film electrode for working and auxiliary electrode (abstract). Thus, Kim teaches both working and counter electrodes are gold electrodes suitable for electrochemical ELISA system.
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 Pt counter electrode in Sokolov to gold counter electrode, as taught by Kim, which teaches a suitable alternative Au counter electrode for electrochemical ELISA system (abstract in Kim).
Claim 15 is rejected under 35 U.S.C. 103 as being unpatentable over Sokolov, as applied to claim 1 above, and further in view of Atrache et al. (US6004766A).
Regarding claim 15, Sokolov teaches the method of claim 1, and further teaches wherein the substrate solution further comprises hydrogen peroxide (H2O2+DAB as shown in Fig.2).
Sokolov is silent to wherein the substrate solution further comprises phosphate-citrate buffer solution.
Atrache teaches an ELISA method. After anti-Salmonella IgG-Horseradish Peroxidase labelled conjugate was added. After 30 minutes incubation at 37 C., the wells were washed three times with TST, then substrate (2,2-azinobis(3-ethylbenzthiazoline sul fonic acid)) in citrate-phosphate buffer (0.1M, pH 4) containing hydrogen peroxide (0.005%) was added (Col. 6 Ln 44-51).
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 substrate solution to further comprise citrate-phosphate buffer containing hydrogen peroxide, as taught by Atrache, which teaches the suitable substrate solution for the ELISA method (Col. 6 Ln 44-51 in Atrache).
Claim 17 is rejected under 35 U.S.C. 103 as being unpatentable over Sokolov, as applied to claim 1 above, and further in view of Sharma et al. (Ultrasound wave-mediated enzyme-linked immunosorbent assay technique, Analytica Chimica Acta, 2009, 650, 241-246).
Regarding claim 17, Sokolov teaches the method of claim 1, and is silent to further comprising vibrating the sample with a vibration exciter to excite at least one of the capture molecule, the plurality of blocking molecules, the sample, the target biomarker, the detection molecule, the detection molecule-target biomarker complex, the sandwich complex, the redox-capable enzyme, or the chromogenic substrate.
Sharma teaches ultrasound wave can dramatically reduce the ELISA timing without losing its specificity or sensitivity. Ultrasound-mediated ELISA was best achieved on an activated microtiter plate which was able to covalently bind antigen or antibody in 10 min when subjected to ultrasoundwaves in a sonicator bath having a temperature of 37◦C, operating at a frequency of 40 KHz and an output power of 120W. Blocking, antibody binding and secondary antibody–enzyme conjugate binding were also accomplished in 10 min each in the sonicator bath under similar conditions (abstract). Thus, Sharma teaches further comprising vibrating the sensor with a vibration exciter (ultrasoundwaves in a sonicator bath) to excite at least one of the capture molecule (sections 2.3-2.4), the plurality of blocking molecules (section 2.4), the sample (section 2.5), the target biomarker (section 2.5), the detection molecule (section 2.6), the detection molecule-target biomarker complex (section 2.6), the sandwich complex (section 2.6), or the redox-capable enzyme (section 2.6).
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 method in Sokolov to further comprise vibrating the sensor with a vibration exciter to excite at least one of the capture molecule, the plurality of blocking molecules, the sample, the target biomarker, the detection molecule, the detection molecule-target biomarker complex, the sandwich complex, or the redox-capable enzyme, as taught by Sharma, since it would dramatically reduce the ELISA timing without losing its specificity or sensitivity (abstract in Sharma).
Conclusion
The prior arts made of record and not relied upon are considered pertinent to applicant's disclosure: Dill et al. (US20070231794A1) teaches an ELISA sensor as shown in Figs. 1-2. Dill (US20200072823A1) teaches an ELISA detection process. Gao et al. (US20060160100A1) teaches an enzymatic electrochemical detection assay.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to SHIZHI QIAN whose telephone number is (571)272-3487. The examiner can normally be reached Monday-Thursday 8:00 am-5:00 pm.
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/SHIZHI QIAN/Primary Examiner, Art Unit 1795
