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 .
Status of the Claims
The Amendment filed August 6, 2026 has been entered. Claims 1-12, 14(w)-15(w), 17(w)-18(w), and 19 have been amended; claims 13, 16 and 20 have been cancelled; and claims 14-15 and 17-18 have been withdrawn. Claims 1-12 and 19 are currently examined herein.
Status of the Rejection
Applicant’s amendments to the specification and drawing do not overcome the specification objection previously set forth in the Non-Final Office Action mailed May 6, 2026.
Applicant’s amendments to the Claims have overcome all claim objections previously set forth in the Non-Final Office Action mailed May 6, 2026.
New grounds of claim objection are necessitated by the amendment as outlined below.
Applicant’s amendments to the Claims have partially overcome the 112(b) rejections previously set forth in the Non-Final Office Action mailed May 6, 2026.
New grounds of rejection under 35 U.S.C. § 112(b) are necessitated by the amendment as outlined below.
Applicant’s amendments to the Claims have overcome all 112(d) rejections previously set forth in the Non-Final Office Action mailed May 6, 2026.
All 35 U.S.C. § 102 and 103 rejections from the previous office action are withdrawn in view of the Applicant’s amendment.
New grounds of rejection under 35 U.S.C. § 103 are necessitated by the amendments as outlined below.
Specification
35 U.S.C. 112(a) or pre-AIA 35 U.S.C. 112, requires the specification to be written in “full, clear, concise, and exact terms.” The specification is replete with terms which are not clear, concise and exact. The specification should be revised carefully in order to comply with 35 U.S.C. 112(a) or pre-AIA 35 U.S.C. 112. Examples of some unclear, inexact or verbose terms used in the specification are: In Fig.9, the amended 3rd equation for calculating the target concentration contradicts to the 2nd equation since the equation for calculating [target] cannot be derived from the 2nd equation. Furthermore, this instant application is similar to applicant’s own journal publication: Nguyen et al. (Label-free, real-time monitoring of cytochrome C drug response in microdissected tumor biopsies with a multi-well aptasensor platform, Science Advances, 2024, 10, eadn5875), wherein the analyte concentration is calculated by equation (2) on page 14, which was derived from equation 1 on page 14. Note that the equation (1) on page 14 of the above journal publication is essentially the same as the 2nd equation in Fig.9 of this instant application. Thus, the equation for calculating the analyte concentration is still not correct.
Claim Objection
Claims 1 and 12 are objected to because of the following informalities:
Claim 1: please amend “E-AB” in Ln 7 to – electrochemical aptamer-based (E-AB)--.
Claim 12: please amend
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to -- K1/2--.
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.
Claim 12 is 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 12, claim 12 recites “the midpoint of the binding curve”, which lacks antecedent basis. Furthermore, claim 12 recites an equation to calculate concentration of the analyte. As explained in the specification objection, the equation contradicts to the 2nd equation in Fig.9. A claim, although clear on its face, may also be indefinite when a conflict or inconsistency between the claimed subject matter and the specification disclosure renders the scope of the claim uncertain as inconsistency with the specification disclosure or prior art teachings may make an otherwise definite claim take on an unreasonable degree of uncertainty. In re Moore, 439 F.2d 1232, 1235-36, 169 USPQ 236, 239 (CCPA 1971); In re Cohn, 438 F.2d 989, 169 USPQ 95 (CCPA 1971); In reHammack, 427 F.2d 1378, 166 USPQ 204 (CCPA 1970) [see MPEP 2173.03]. Thus, the scope of claim 12 is indefinite.
Claim Rejections - 35 USC § 103
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claims 1-2, 4-5, 11 and 19 are rejected under 35 U.S.C. 103 as being unpatentable over Plaxco et al. (US20210196161A1) and in view of Santos-Cancer et al. (Direct, real-time detection of adenosine triphosphate release from astrocytes in three-dimensional culture using an integrated electrochemical aptamer-based sensor, ACS Chemical Neuroscience, 2019, 10, 2070-2079). Mirceski et al. (Square-wave voltammetry, ChemTexts, 2018, 4, 17) is an evidence for claim 11.
Regarding claim 1, Plaxco teaches a system ( an electrochemical sensing system [para. 0028, 0090]) configured to measure a concentration of an analyte (measuring the concentration of a target species in a sample by the use of an electrochemical sensor [para. 0091; claim 1]; thus the electrochemical sensing system is configured to perform the intended use of measuring a concentration of an analyte), the system comprising:
An integrated E-AB sensor (an E-AB sensing platform as shown in Fig.1A [para. 0009]) comprising:
an aptamer operably connected to a conductive substrate (an aptamer 102 bound to an electrode substrate 101 in Fig.1A [para.0009]);
one or more controllers comprising logic that, when executed by the one or more controllers, causes the system to perform operations (non-transitory computer-readable recording media having stored thereon data and/or an encoding program that causes a computer to execute series of operations. wherein the data and/or series of operations causes the operation of an electrochemical sensing system to effect the methods of the invention. The non-transitory computer-readable recording media may effect operations such as: controlling a potentiostat or equivalent device to deliver a series of stepped voltage pulses to a deployed electrochemical sensor; controlling a data collection device to record sensor current outputs following the delivery of voltage pulses; controlling a processor to perform calculations that derive one or more selected measures of current decay from sensor output data; storing a calibration curve that relates a selected measure of current decay to target concentration; controlling a processor to calculate target concentration based on sensor outputs and a stored calibration curve; storing instructions for the performance of the methods of the invention; and other operations of the invention [para.0087-0088]) comprising:
conducting square wave voltammetry (SWV) to apply an electric current to the conductive substrate (Fig.1B depicts a representative square wave voltammetry trace for this type of E-AB sensor wherein peak current is higher in the presence than in the absence of target [para. 0009]; All SWV measurements were performed using a three-electrode setup and with a CH Instruments electrochemical workstation [para. 0121]; the non-transitory computer-readable recording media may effect operations such as: controlling a potentiostat or equivalent device to deliver a series of stepped voltage pulses to a deployed electrochemical sensor [para. 0088]); and
conducting kinetic differential measurement (KDM) (non-transitory computer-readable recording media having stored thereon data and/or an encoding program that causes a computer to execute series of operations [para.0087]; Kinetic differential measurements correction techniques applied to the measurements provide a corrected signal [para. 0014]) to determine the concentration of the analyte based on changes in the electric current caused by conformational changes in the aptamer upon an interaction between the aptamer and the analyte (calculating the concentration of the target species by application of a mathematical relationship between selected measure of current decay and the concentration of the target species in the sample [claim 1]; computer programs, software, and operations which enable the acquisition and interpretation of electrochemical sensor outputs to measure target concentration [para. 0008]; aptamer undergoes a conformational change upon target binding [para. 0097, 0105; Fig.1A]; signals via a binding-induced conformational change that alters electron transfer from the reporter, leading in turn to an easily measurable electrochemical output [para. 0109; Fig.1B]).
Plaxco is silent to the system further comprising: a multielectrode cell culture platform comprising a sensor layer and a wells layer disposed over the sensor layer, wherein the wells layer comprises one or more wells sized and shaped for culturing an intact tissue.
Santos-Cancer teaches a multielectrode cell culture platform comprising a sensor layer (a sensor layer having an E-AB sensor comprising Pt CE, Pt RE and Au WE as shown in Scheme 1) and a wells layer disposed over the sensor layer (a wells layer comprising a 3D cell culture well disposed on the sensor layer as shown in Scheme 1), wherein the wells layer comprises one or more wells sized and shaped for culturing an intact tissue (one 3D cell culture well as shown in scheme 1 is sized and shaped for culturing an intact tissue [E-AB sensors detect ATP when interfaces with a collagen tissue scaffold]).
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 system in Plaxco by providing a multielectrode cell culture platform comprising a sensor layer and a wells layer disposed over the sensor layer, wherein the wells layer comprises one or more wells sized and shaped for culturing an intact tissue, as taught by Santos-Cancer, since it would interface the E-AB sensor with a 3D cell culture well for the real-time and continuous detection of biological tissue (the 3rd paragraph in Col.2 on page 2071 in Santos-Cancer).
Regarding claim 2, Modified Plaxco teaches the system of claim 1, and Plaxco teaches wherein:
the integrated E-AB sensor further comprises a redox reporter (methylene blue redox reporter 103 in Fig.1A [para. 0009]) configured to modify the electric current to and/or from the conductive substrate of the integrated E-AB sensor (When target 105 binds to the aptamer, the resulting conformation change alters the proximity of the reporter 103 to the substrate 101, increasing the rate of electron transfer 104, and Fig.1B depicts a representative SWV for this type of E-AB sensor wherein peak current is higher in the presence than in the absence of target [para. 0009; Figs.1A-1B ]; thus the MB redox reporter is configured to perform the claimed function above); and
the aptamer comprises a nucleic acid aptamer (aptamer 102 in Fig.1A [para.0009]; the aptamer may comprise a DNA aptamer, RNA aptamer, or an aptamer comprising non-natural nucleic acids [para. 0022]) comprising a polynucleotide sequence (nucleic acid aptamer inherently comprises a polynucleotide sequence) configured to interact with the analyte (target 105 binds to the aptamer [para. 0009]), wherein an interaction between the polynucleotide sequence and the analyte causes a conformational change in the nucleic acid aptamer, a positional change in the redox reporter, and a change in the electric current of the integrated E-AB sensor (When target 105 binds to the aptamer, the resulting conformation change alters the proximity of the reporter 103 to the substrate 101, increasing the rate of electron transfer 104 [para. 0009; Figs. 1A-1B]; current flows are altered by target binding and are assessed by voltammetric methods including SWV [para. 0016]);
wherein the electric current is applied with the SWV (All SWV measurements were performed using a three-electrode setup and with a CH Instruments electrochemical workstation [para. 0121]; thus the CHI electrochemical workstation is configured to apply the electric current) and the change in the electric current is processed with the KDM to determine the concentration of the analyte (non-transitory computer-readable recording media having stored thereon data and/or an encoding program that causes a computer to execute series of operations [para. 0087]; calculating the concentration of the target species by application of a mathematical relationship between selected measure of current decay and the concentration of the target species in the sample [claim 1]; the scope of the invention encompasses computer programs, software, and operations which enable the acquisition and interpretation of electrochemical sensor outputs to measure target concentration [para. 0008]; Kinetic differential measurements correction techniques applied to the measurements provide a corrected signal [para. 0014]).
Regarding claim 4, modified Plaxco teaches the system of claim 2, and Plaxco teaches wherein the nucleic acid aptamer comprises deoxyribonucleic acid (DNA) (DNA aptamer [para. 0022]) and the polynucleotide sequence comprises a DNA sequence (DNA aptamer inherently comprises a DNA sequence).
Regarding claim 5, modified Plaxco teaches the system of claim 2, and Plaxco teaches wherein the redox reporter comprises methylene blue (MB) (methylene blue redox reporter 103 in Fig.1A [para. 0009]).
Regarding claim 11, modified Plaxco teaches the system of claim 1, and Plaxco teaches wherein the conducting SWV uses a combined square wave applied to the conductive substrate of the integrated E-AB sensor (For the SWV measurements, the sensors were interrogated from 0.0 V to −0.5 V versus Ag/AgCl, using an amplitude of 50 mV, potential step sizes of 1-5 mV, and varying frequencies from 10 Hz to 500 Hz. All SWV measurements were performed using a three-electrode setup and with a CH Instruments electrochemical workstation [para. 0121]. As evidenced by Mirceski, SWV does use a combined square wave as shown in Fig.5b. Thus, the CHI electrochemical workstation is configured to use a combined square wave applied to the conductive substrate of the integrated E-AB sensor).
Regarding claim 19, Plaxco teaches a kit (kits and systems [para. 0090]) for measurement of an analyte (for calculating target concentration [para. 0090-0091]; an electrochemical sensor is any sensor that is capable of measuring the concentration of a target species in a sample [para. 0021]), the kit comprising:
a system comprising an integrated E-AB sensor (an electrochemical sensing system comprising an E-AB sensor in Fig.1A [para. 0009, 0028]),
wherein the integrated E-AB sensor comprises:
an aptamer operably connected to a conductive substrate of the E-AB sensor (an aptamer 102 bound to an electrode substrate 101 in Fig.1A [para.0009]); and
one or more controllers comprising logic that, when executed by the one or more controllers, causes the system to perform operations (non-transitory computer-readable recording media having stored thereon data and/or an encoding program that causes a computer to execute series of operations. wherein the data and/or series of operations causes the operation of an electrochemical sensing system to effect the methods of the invention. The non-transitory computer-readable recording media may effect operations such as: controlling a potentiostat or equivalent device to deliver a series of stepped voltage pulses to a deployed electrochemical sensor; controlling a data collection device to record sensor current outputs following the delivery of voltage pulses; controlling a processor to perform calculations that derive one or more selected measures of current decay from sensor output data; storing a calibration curve that relates a selected measure of current decay to target concentration; controlling a processor to calculate target concentration based on sensor outputs and a stored calibration curve; storing instructions for the performance of the methods of the invention; and other operations of the invention [para.0087-0088]) comprising:
conducting SWV to apply an electric current to the conductive substrate (Fig.1B depicts a representative square wave voltammetry trace for this type of E-AB sensor wherein peak current is higher in the presence than in the absence of target [para. 0009]; All SWV measurements were performed using a three-electrode setup and with a CH Instruments electrochemical workstation [para. 0121]; the non-transitory computer-readable recording media may effect operations such as: controlling a potentiostat or equivalent device to deliver a series of stepped voltage pulses to a deployed electrochemical sensor [para. 0088]); and
conducting KDM (non-transitory computer-readable recording media having stored thereon data and/or an encoding program that causes a computer to execute series of operations [para.0087]; Kinetic differential measurements correction techniques applied to the measurements provide a corrected signal [para. 0014]) to determine a concentration of the analyte based on a change in the electric current caused by a conformational change in the aptamer upon an interaction between the aptamer and the analyte (calculating the concentration of the target species by application of a mathematical relationship between selected measure of current decay and the concentration of the target species in the sample [claim 1]; computer programs, software, and operations which enable the acquisition and interpretation of electrochemical sensor outputs to measure target concentration [para. 0008]; aptamer undergoes a conformational change upon target binding [para. 0097, 0105; Fig.1A]; signals via a binding-induced conformational change that alters electron transfer from the reporter, leading in turn to an easily measurable electrochemical output [para. 0109; Fig.1B]); and
instructions for a use of the kit in a method for measuring the analyte with the kit (instructions for performing the methods of the invention [para. 0090]; [para. 0048] details a method of measuring the concentration of a target species in a sample by the use of an E-AB sensor).
Plaxco is silent to the system further comprising: a multielectrode cell culture platform comprising a sensor layer and a wells layer disposed over the sensor layer, wherein the wells layer comprises one or more wells sized and shaped for culturing an intact tissue.
Santos-Cancer teaches a multielectrode cell culture platform comprising a sensor layer (a sensor layer having an E-AB sensor comprising Pt CE, Pt RE and Au WE as shown in Scheme 1) and a wells layer disposed over the sensor layer (a wells layer comprising a 3D cell culture well disposed on the sensor layer as shown in Scheme 1), wherein the wells layer comprises one or more wells sized and shaped for culturing an intact tissue (one 3D cell culture well as shown in scheme 1 is sized and shaped for culturing an intact tissue [E-AB sensors detect ATP when interfaces with a collagen tissue scaffold]).
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 system in Plaxco by providing a multielectrode cell culture platform comprising a sensor layer and a wells layer disposed over the sensor layer, wherein the wells layer comprises one or more wells sized and shaped for culturing an intact tissue, as taught by Santos-Cancer, since it would interface the E-AB sensor with a 3D cell culture well for the real-time and continuous detection of biological tissue (the 3rd paragraph in Col.2 on page 2071 in Santos-Cancer).
Claim 3 is rejected under 35 U.S.C. 103 as being unpatentable over Plaxco and Santos-Cancer, as applied to claim 1 above, and in view of Pluznick et al. (US20190064161A1).
Regarding claim 3, modified Plaxco teaches the system of claim 1, and is silent to wherein a form factor of the integrated E-AB sensor comprises a dip-stick form factor.
Pluznick teaches an E-AB sensor comprises a dip-stick form factor (One preferred ligand binding part of Olfr90 is an aptamer representing the binding site of Olfr90 and the aptamer is labeled with a tag, a fluorophore, for example. An Olfr90, or the ligand binding part thereof, such as an aptamer is preferably attached to a surface such as a strip or a dip stick [para. 0008; claim 17]).
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 E-AB sensor to comprise a dip-stick form factor, as taught by Pluznick, since Pluznick teaches an alternative suitable configuration of the E-AB sensor having the aptamer attached to a surface such as a dip stick [para. 0008; claim 17].
Claims 6-7 are rejected under 35 U.S.C. 103 as being unpatentable over Plaxco and Santos-Cancer, as applied to claim 2 above, and in view of Plaxco et al. (WO2020252401A1; hereinafter Plaxco’401).
Regarding claims 6-7, modified Plaxco teaches the system of claim 2, and Plaxco teaches wherein the redox reporter is linked to the nucleic acid aptamer (an aptamer 102 is modified with a methylene blue redox reporter 103 [para. 0009]; Fig.1A shows the MB redox reporter is linked to the distal end of the aptamer 102).
Plaxco does not explicitly teaches wherein the redox reporter is linked to the nucleic aptamer “at a terminus of the nucleic acid aptamer” (of claim 6); and wherein the terminus is a 3′ terminus of the nucleic acid aptamer (of claim 7).
Plaxco’401 teaches an E-AB sensor (claims 1 and 4; E-AB sensor [para. 0100]), wherein the aptamer is functionalized with -SH at the 5’ end for attachment to the SAM functionalized gold electrode surface and functionalized at the 3’ end with methylene blue [para. 0026]. Thus, Plaxco’401 teaches wherein the redox reporter of methylene blue is linked to the nucleic acid aptamer at a 3’ terminus of the nucleic acid aptamer.
Given the teachings of Plaxco regarding methylene blue (MB) redox reporter is linked to the nucleic acid aptamer at the distal end (see Fig.1A), and the teachings of Plaxco’401 regarding methylene blue is linked to the nucleic acid aptamer at a 3’ terminus of the nucleic acid aptamer, 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 E-AB sensor of modified Plaxco to have the MB redox reporter linked to the nucleic acid aptamer at a 3’ terminus of the nucleic acid aptamer, as taught by Plaxco’401, since Plaxco’401 teaches it would be suitable to functionalize the aptamer at a 3’ end for attaching the MB redox reporter [para. 0026].
Claims 8-9 are rejected under 35 U.S.C. 103 as being unpatentable over Plaxco and Santos-Cancer, as applied to claim 2 above, and in view of Poturnayova et al. (Optimization of cytochrome c detection by acoustic and electrochemical methods based on aptamer sensors, Sensors and Actuators B: Chemical, 2017, 238, 817-827).
Regarding claims 8-9, modified Plaxco teaches the system of claim 2, and is silent to: (1) wherein the analyte comprises Cytochrome C (Cyt-C) and the polynucleotide sequence comprises a DNA sequence specific for interaction with Cyt-C (of claim 8); and (2) wherein the analyte comprises Cyt-C and the polynucleotide sequence comprises a DNA sequence specific for interaction with Cyt-C, wherein the DNA sequence specific for interaction with Cyt-C comprises a DNA sequence that is at least 80% identical to SEQ ID NO:1 (of claim 9).
Poturnayova teaches an E-AB sensor for the detection of Cyt-C (title and abstract), wherein the aptamer 40 specific for interaction with Cyt-C comprises a DNA sequence:
5′- CC GTG TCT GGG GCC GAC CGG CGC ATT GGG TAC GTT GTT GC-3′ (see apt 40 in Fig.1). Note that the SEQ ID NO:1 of this instant application is: 5′-Thio-C6-Disulfide/CC GTG TCT GGG GCC GAC CGG CGC ATT GGG TAC GTT GTT GC/—NH2-3′ [para. 0016 ] in PG-Pub of this instant specification. Thus, the disclosed DNA sequence of the aptamer 40 is essentially the same as the claimed SEQ ID NO:1 except the modifiers at the 5’ terminus and the 3’ terminus.
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 nucleic acid aptamer in modified Plaxco with the aptamer having the DNA sequence of 5′- CC GTG TCT GGG GCC GAC CGG CGC ATT GGG TAC GTT GTT GC-3′ and take the modified E-AB sensor for detecting analyte of Cyt-C, as taught by Poturnayova, since it would allow to detect the analyte of Cyt-C, which takes part in essential biological functions as electron carrier and metal-ion liaison (Introduction in Poturnayova).
With the substituted aptamer as the aptamer specific for detecting Cyt-C as the analyte, modified Plaxco teaches: (1) wherein the analyte comprises Cytochrome C (Cyt-C) and the polynucleotide sequence comprises a DNA sequence specific for interaction with Cyt-C (claim 8); and (2) wherein the analyte comprises Cyt-C and the polynucleotide sequence comprises a DNA sequence specific for interaction with Cyt-C, wherein the DNA sequence specific for interaction with Cyt-C comprises a DNA sequence that is at least 80% identical to SEQ ID NO:1 (claim 9).
Claim 10 is rejected under 35 U.S.C. 103 as being unpatentable over Plaxco and Santos-Cancer, as applied to claim 2 above, and in view of Poturnayova et al. (Optimization of cytochrome c detection by acoustic and electrochemical methods based on aptamer sensors, Sensors and Actuators B: Chemical, 2017, 238, 817-827), Catanante et al. (Sensitive analytical performance of folding based biosensor using methylene blue tagged aptamers, Talanta, 2016, 153, 138-144), Kelley et al. ( US20230107004A1), and McHenry et al. (Voltammetry peak tracking for longer-lasting and reference-electrode-free electrochemical biosensors, Biosensors, 2022, 12, 782).
Regarding claim 10, modified Plaxco teaches the system of claim 2, and is silent to wherein the analyte comprises Cyt-C and the polynucleotide sequence comprises a DNA sequence specific for interaction with Cyt-C, wherein the DNA sequence specific for interaction with Cyt-C is SEQ ID NO:1.
Poturnayova teaches an E-AB sensor for the detection of Cyt-C (title and abstract), wherein the aptamer 40 specific for interaction with Cyt-C comprises a DNA sequence: 5′- CC GTG TCT GGG GCC GAC CGG CGC ATT GGG TAC GTT GTT GC-3′ (see apt 40 in Fig.1). Note that the SEQ ID NO:1 of this instant application is: 5′-Thio-C6-Disulfide/CC GTG TCT GGG GCC GAC CGG CGC ATT GGG TAC GTT GTT GC/—NH2-3′ [para. 0016 ] in PG-Pub of this instant specification. Thus, the disclosed DNA sequence of the aptamer 40 is essentially the same as the claimed SEQ ID NO:1 except the modifiers at the 5’ terminus and the 3’ terminus.
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 nucleic acid aptamer in modified Plaxco with the aptamer having the DNA sequence of 5′- CC GTG TCT GGG GCC GAC CGG CGC ATT GGG TAC GTT GTT GC-3′ and take the modified E-AB sensor for detecting analyte of Cyt-C, as taught by Poturnayova, since it would allow to detect the analyte of Cyt-C, which takes part in essential biological functions as electron carrier and metal-ion liaison (Introduction in Poturnayova).
With the substituted aptamer as the aptamer specific for detecting Cyt-C as the analyte, modified Plaxco teaches: wherein the analyte comprises Cyt-C and the polynucleotide sequence comprises a DNA sequence specific for interaction with Cyt-C, wherein the DNA sequence specific for interaction with Cyt-C is essentially the same as SEQ ID NO:1 EXCEPT the modifiers at the 5’ terminus and the 3’ terminus.
Modified Plaxco is silent to wherein the modifier at the 5’ terminus is Thio-C6-Disulfide, and the modifier at the 3’ terminus is NH2.
Plaxco further teaches methylene blue (MB) redox reporter 103 is linked to the aptamer 102 at its distal end, and the other end of the aptamer 102 is linked to the electrode substrate 101 ([para. 0009] and Fig.1A).
Catanante teaches E-AB sensor comprising methylene blue as the redox reporter (title and abstract). Typically, the aptamer is modified at the 5’-end for electrode attachment (e.g., thoilated) and at the distal, 3’-end, with a redox probe molecule such as methylene blue (MB) (the first paragraph in Introduction on page 138).
Kelley teaches an E-AB sensor comprising a receptor (which is an aptamer) and a redox reporter (abstract; claims 1, 26-27 and [para. 0035]), and one of the probe sequences is 5′-SH-MC6-TAC CAG CTA TTG TAT CTA ATA AGA-NH2-3′ [para. 0157, 0171]. The redox report is methylene blue ( claim 16; a methylene blue reporter molecule was conjugated to a ssDNA probe [para. 0216]). Thus, Kelley teaches an E-AB sensor wherein the redox reporter is MB, and the modifier of the aptamer at the 3’ terminus is NH2, and the 5’ end of the aptamer is thoilated containing SH group.
Given the teachings of Plaxco regarding MB redox reporter is linked to the aptamer at its distal end; the teachings of Catanante regarding aptamer being typically modified at the 3’-end for MB redox probe and the 5’ end of the aptamer being modified with a modifier containing -SH group for electrode attachment; and the teachings of Kelley regarding the MB redox reporter and the aptamer modified at the 3’-end with an amino modifier (i.e., NH2) and modified at the 5’ end with a modifier containing a thiol group -SH, 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 3’ terminus of the aptamer in modified Plaxco with an amino modifier to provide NH2 at the 3’ terminus, as taught by combined Plaxco, Catanante, and Kelley, since it would allow to attach the MB redox reporter at the 3’ terminus of the aptamer. Note that the 5’ end containing a thiol group -SH would be used for electrode attachment since Catanante teaches the aptamer is modified at the 5’-end for electrode attachment (e.g., thoilated).
Modified Plaxco is silent to wherein the modifier at the 5’ terminus is Thio-C6-Disulfide.
McHenry teaches electrochemical aptamer-based sensor (abstract and Fig.1) comprising aptamer, [/5ThioMC6-D/CGAGG GTACC GCAAT AGTAC TTATT GTTCG CCTAT TGTGG GTCGG/3MeBlN/] with conjugated methylene blue and thiol ends (section 2.1). Fig.1 shows one end of the aptamer is attached to the electrode and the distal end of the aptamer is linked to the redox reporter. Since the 3’-end is conjugated with methylene blue redox reporter, the 5’-end of the aptamer is then attached to the electrode. Thus, McHenry teaches wherein the aptamer is modified at the 5’-end with a modifier of ThioMC6-D for attaching the aptamer to the electrode.
Given the teachings of Catanante regarding the aptamer is typically modified at the 5’-end for electrode attachment (e.g., thoilated); and the teachings of McHenry regarding the aptamer is modified at the 5’-terminus with a modifier of ThioMC6-D for attaching the aptamer to the electrode , it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention modify the 5’-terminus of the aptamer in modified Plaxco with a modifier of ThioMC6-D, as taught by combined Catanante and McHenry, since it would allow to attach the aptamer to the electrode at the 5’-terminus.
With the above modifications of the modifiers at the 3’ terminus and the 5’-terminus, the modified aptamer has the DNA sequence of SEQ ID: No:1.
Claim 12 is rejected under 35 U.S.C. 103 as being unpatentable over Plaxco and Santos-Cancer, as applied to claim 1 above, and in view of Downs et al. (Improved calibration of electrochemical aptamer-based sensors, Scientific reports, 2022, 12, 5535).
Regarding claim 12, modified Plaxco teaches the system of claim 1, and is silent to wherein the conducting KDM to determine the concentration of the analyte is configured to calculate the concentration of the analyte according to:
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90
426
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wherein:
[Analyte] is the concentration of the analyte, optionally in ng/mL;
KDM is the KDM value observed at the concentration of the analyte;
KDMmin is KDM observed in absence of the analyte;
KDMmax is KDM expected at saturation of the analyte;
nH is Hill coefficient; and
K1/2 denotes a midpoint of a binding curve.
.
Downs teaches an E-AB sensor (title, abstract and Fig.1), and also teach KDM to determine analyte concentration (see Fig.1 and equations 1-2) using the equation (2):
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94
656
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Note that the above equation (2) for calculating the target analyte concentration is derived from the equation (1):
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74
618
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The above equation (1) is the same as the 2nd equation in Fig.9 of this instant application. Thus, Downs teaches the equation (2), which is derived from the 2nd equation in Fig.9 of this instant application, to calculate the analyte concentration.
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 computer program(s)/software in the computer of modified Plaxco to calculate the concentration of the analyte according to the equation (2) of Downs, since it would allow to calculate the target concentration based on the E-AB sensor output (see Figs. 1D to 1E; the first paragraph on -page 2 in Downs).
Note that the amended equation to calculate the target concentration cannot be derived from the second equation in instant Fig.9. The equation (1) in Downs is the same as the second equation in Fig.9 of this instant specification.
Response to Arguments
Applicant's arguments, see Remarks Pgs. 10-19, filed 8/6/2026, with respect to the specification objection and 35 U.S.C. § 112(b), 102 and 103 rejections have been fully considered.
Applicant’s Argument #1:
Regarding specification objection, Applicant argues at pages 10-11 that the equation for calculating the target concentration has been amended.
Examiner’s Response #1:
Applicant’s arguments have been fully considered, but the amended equation cannot be derived from the 2nd equation in Fig.9 of the instant specification.
Applicant’s Argument #2:
Regarding claim rejection under 112(b) for claim 12, applicant argues at pages 12-13 that the equation is amended.
Examiner’s Response #2:
Applicant’s arguments have been fully considered, but the amended equation cannot be derived from the 2nd equation in Fig.9 of the instant specification.
Applicant’s Argument #3:
Regarding the 102 and 103 rejections, applicant argues at pages 14-18 that the prior art does not teach the amended features: a multielectrode cell culture platform comprising a sensor layer and a wells layer disposed over the sensor layer, wherein the wells layer comprises one or more wells sized and shaped for culturing an intact tissue.
Examiner’s Response #3:
Applicant’s arguments have been fully considered, but are moot in view of the new grounds of rejection for amended claims 1 and 19 above.
Conclusion
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Luan V Van can be reached on 571-272-8521. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/SHIZHI QIAN/Primary Examiner, Art Unit 1795
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