DETECTION ASSAY

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 . DETAILED ACTION 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. Status of the claims Claims 1-18 are examined on merits in this office action. 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 of this title, 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, 3-7, 10-11 and 12 are rejected under 35 U.S.C. 103 as obvious over Li et al. (Langmuir 2019; Cite # 2, in the IDS of 9/28/2023) in view of Badescu et al (Bioconjugate Chem. 2014). In regard to claims 1, 3-7 and 12, Li discloses a surface having plurality of reaction zones PNG media_image1.png 98 159 media_image1.png Greyscale for immobilization of his tag protein (6His-anti-HER2 antibody) wherein the surface is provided by utilizing sulfone based bifunctional linker (abstract). Li teaches that His-tagged protein immobilization via surface vinyl sulfone­imidazole coupling on solid supports lead to improved sensitivity of biosensors, as the protein is oriented in a more favorable way. Li teaches that the surface with immobilized sulfone provides site-directed immobilization of proteins (biological molecules) by means of a sulfone based cross-linking agent (= VS surface) that attaches the His tag Ab to the substrate (Au surface = substrate) via a functionalized EG6-OH SAM (= linker; figure 1 a, figure 5b). Such immobilization via surface vinyl sulfone-imidazole coupling on solid supports lead to improved sensitivity of biosensors, as the protein is oriented in a more favorable way (see "conclusions" on page 16473; bridging paragraph between pages 16471-16472). Li does not disclose the use of a sulphone-based cross-linker having the structure of Formula (1 ) as claimed. Badescu discloses trifunctional linker 4-[2,2-Bis[(p-tolylsulfonyl)methyl]acetyl) benzoic acid-NHS ester; bis-sulfone NHS; i.e PNG media_image2.png 239 284 media_image2.png Greyscale ; page 1133, 1st col) for providing conjugates of antibody with a drug molecule (Abstract). Badescu teaches that the linker is useful for providing covalent attachment of drug to targeting antibody (Introduction). Badescu teaches that the bis-reactive sulfone group (bis-alkylating group) is capable of undergoing reaction with both sulfur atoms derived from a reduced disulfide bond in antibodies and antibody fragments resulting in covalent re-bridging via a three-carbon bridge leaving the protein (antibody) structurally intact and the resulting conjugates are stable and retained antigen-binding (Abstract and introduction). Badescu teaches that the three-carbon bridge is small enough and sufficiently flexible not to perturb the tertiary structure of the antibody (page 1126, 1st col.). PNG media_image3.png 239 723 media_image3.png Greyscale Badescu teaches that the linker with the antibody can be conjugated to drugs to provide antibody-drug conjugate (ADC) to the fluorophores to provide fluorescent tagged antibody (Fir 2). PNG media_image4.png 477 895 media_image4.png Greyscale The trifunctional linker is of the same trifunctional linker as claimed (see claim 1 and formula 4 of claim 4). Note that the trifunctional linker of Badescu comprises -NH-for R3 which is end product of reaction of NHS group with amine containing surfaces. Note that it is the final product of the substrate wherein biological molecule is immobilized to the substrate through the linker and the linker after reaction with the biomolecule and the substrate will not retain the functional groups as described by R2 and R1. Therefore, from the description in mind of Li and Badescu, it would be obvious to ordinary skilled in the art to easily envisage considering the trifunctional linker of Badescu for surface immobilization of antibody in view of Li with the expectation of expanding the arsenal of linkers and immobilization processes of various antibodies with other linkers, in the method of Li with a reasonable expectation of success. One of ordinary skilled in the art from the reading would understand that for the sulfone linker of Li, the antibody needs to be tagged with 6His but however, the linker of Badescu, as described above, does not require tagging antibody with 6His because Badescu teaches that the bis-reactive sulfone group is capable of undergoing reaction with both sulfur atoms derived from a reduced disulfide bond in antibodies and antibody fragments resulting in covalent re-bridging via a three carbon bridge leaving the antibody structurally intact and the resulting conjugates are stable and retained antigen-binding. Badescu also teaches that monoalkylation reagents leave the original disulfides unbridged, potentially introducing instability to the antibody (Introduction). Therefore, one of ordinary skilled in the art would be motivated to utilize that bis-alkylating group of the linker of Badescu for binding to various antibodies with ease with a reasonable expectation of success. Moreover, since Badescu teaches drugs or fluorophores can be conjugated with the antibody by utilizing the other functional group of the linker as shown in Fig,2, one of ordinary skilled in the art can easily envisage the other functional group (including amine containing surfaces) for immobilization of the linker to discrete surface locations for immobilization of the antibody to the surfaces in view of Li with a reasonable expectation of success. Badescu discloses that the other functional group can be modified with various functional groups including activated form (e.g. NHS) and thus various surfaces can be utilized with appropriate modification for immobilization of antibody utilizing the linker of Badescu with a reasonable expectation of success. In regards to claim 10, Li teaches surface wherein titanium and gold sequentially deposited onto silicon wafers (page 16468, 2nd para of 1st col.), thus disclosing substrate comprising silicon. In regards to claim 11, Li teaches surfaces of surface plamon resonance (SPR) chips, which would be considered as bio-chip surface (page 16466, 1st col.). Claims 1-7 and 10-18 are rejected under 35 U.S.C. 103 as being obvious over Li et al. (Langmuir 2019) in view of Badescu et al (Bioconjugate Chem. 2014) as applied to claims 1, 3-7, 10-11 and 12 above, and further in view of Mcdonnell et al (2016/0287718A1; cited in IDS of 9/28/2023). Li in view of Badescu has been described above providing obviousness of utilizing the bis-alkylating linker of Badescu for immobilization of antibody to biosensor surface for detection of analyte. Li in view of Badesco, as described above, teach gold and titanium surface but however, do not teach utilization of various other commonly used surfaces useful for biosensors and sensor arrays including epoxy surfaces and do not specifically teach a detection process. In regards to claims 2, Mcdonnell teaches conjugate of a specific binding molecule (domain I β2GP1) with PEG polymer (Abstract) via a linker wherein the preferred linker is PNG media_image5.png 133 467 media_image5.png Greyscale PNG media_image6.png 101 448 media_image6.png Greyscale (para [0081]). Mcdonnell teaches that the linker is conjugated to the linker via two cysteine residues derived from a disulfide bridge in the domain I β2GP1 polypeptide (Abstract; claim 16 and para [0137]). Mcdonnell teaches that domain I β2GP1 polypeptide bind to analyte anti- β2GP1 antibody for detection of the analyte (para [0033] and [0037]). Mcdonnell teaches that to use the domain I β2GP1 polypeptide conjugate of the Invention in the detection of antibodies in an individual, an immunoassay may be conducted, the domain I β2GP1 polypeptide conjugate is provided as a reagent, and the antibody is the target in the biological sample (para [0039]). Mcdonnell teaches that in the methods of the invention, domain I β2GP1 polypeptide conjugates of the Invention may be immobilized, by known techniques, onto a suitable solid phase, such as affinity column packing material, or a plastic surface such as a microtiter plate or a dipstick (para [0041]). Therefore, given the fact that surface immobilization with sulfone-based linker useful and common in the art (Li and Mcdonnell) and given the fact that Mcdonnell envisaged immobilization of binding partner to surface utilizing linker having bis-alkylating group for immunoassay detection of analytes and given the fact that various surfaces, including silane surfaces can be utilized, it would be obvious to one of ordinary skilled to easily envisage utilizing the linker having bis-alkylating groups (e.g. PNG media_image2.png 239 284 media_image2.png Greyscale ) as disclosed by Badesco and Mcdonnell on various surfaces, as for example, epoxy silane surface and silane surface modified with various function group, with the expectation of immobilization of antibody on the surfaces for immunoassay detections with a reasonable expectation of success. Moreover, Badesco discloses derivation of the functional groups with amine group and one of ordinary skilled in the art understands that epoxy silane surface having and amine gorup will react with the NHS group of the linker to form a covalent linkage. Moreover, since the basic concept of utilizing the linker has been found to be obvious in view of Li and Badescu and various surfaces for immobilization have been taught by Mcdonnell, various surfaces having various functional groups and derivation of the linker functional group with various group for optimized conjugation would be obvious and within the purview of one of ordinary skilled in the art. In regards to method process of detection as claimed in claims 13-18, Mcdonnell teaches immunoassay detection involving contacting the antibody conjugate with a sample containing the analyte , removing the unbound analyte and detecting the bound analyte with a detectable label. Mcdonnell also teaches competitive immunoassays utilizing labeled antibody (labeled analyte) to compete with the analyte present in the sample for binding to conjugate antibody wherein the amount of label negatively corelates with the amount of analyte (anti-β2GP1) present in the sample. Mcdonnell teaches detectably label includes enzyme label. (para [0043]; [0148]). Mcdonnell teaches various samples, including plasma. Therefore, from the description in mind of Mcdonnell, one of ordinary skilled in the art can envisage that the solid support based immunoassay with the linker immobilized antibody would involve similar contacting, removing and detecting step with labeled antibody or labeled analyte and various samples and various types of common and known immunoassay method steps for detection of the various analytes with the immobilized linker would be obvious to one of ordinary skilled in the art. Claims 1-18 are rejected under 35 U.S.C. 103 as being obvious over Li et al. (Langmuir 2019) in view of Badescu et al (Bioconjugate Chem. 2014) and Mcdonnell et al (2016/0287718A1) as applied to claims 1-7 and 10-18 above, and further in view of Fitzgerald et al (WO2017/085509A1). Li in view of Badescu and Mcdonnell have been described above providing obviousness of utilizing the bis-alkylating linker of Badescu for immobilization of antibody to biosensor surface for detection of analyte utilizing various immunoassay processes. Li in view of Badesco and Mcdonnell as described above, teach biosensor surface of gold and titanium having plurality of reaction zones for immobilization of antibody but however, do not teach utilization of various other commonly used surfaces useful for biosensors and sensor arrays having masking/coating material on the surface wherein the reaction zones are uncoated and also does not teach the masking material further comprises a pigment. In regards to claims 8-10, Fitzgerald teaches biosensor substrates (e.g. microarray) having plurality of reaction zones (page 1 and claims 1 and 10) wherein the substate comprises a coating of a masking material, and plurality of discrete reaction zones onto which one or more binding agents are intended to be covalently attached and wherein zones are uncoated areas on the substrate (page 2, lines 5-8). Fitzgerald teaches that it has surprisingly been found that the coated substrate provides improved detection sensitivity and image quality. Fitzgerald teaches the coated areas of the substrate reduce non-specific binding and the cross-linking of the spots and so background noise is reduced resulting in an enhanced signal-to-noise ratio (page 2, lines 10-14; page 3, line 20 to page 4, line 6). Fitzgerald teaches various types of substates including metal oxides, ceramic, glass and plastic (page 3, line 20 to page 4, line 6) and various types of resin for coating including acrylics and teaches that the coating may contain any suitable ink solvent and/or ink additive (page 4, lines 27-34 and claims 1,10-12, 13, 23 and 24). Fitzgerald further teaches that the masking material has a contact angle of 60-120⁰C (claim 13). Therefore, given the fact that coating/masking, preferably with coating composition comprising black pigment in the area outside of reaction zones are useful for providing biosensor surfaces including microarray surfaces that provides improved detection sensitivity and image quality by reducing background noise resulting in an enhanced signal-to-noise ratio (Fitzgerald), it would be obvious to one of ordinary skilled in the art to easily envisage utilizing the coating composition and various sensor substrates in the method of detection as found obvious in view of Li, Badescu and McDonnell with a reasonable expectation of success. Response to argument Applicants’ arguments and amendments filed 08/21/2026 have been fully considered and are persuasive to overcome the objections in view of the amendments but they are not found persuasive to overcome the rejections under 35 USC 103. Moreover, Applicants’ amendments necessitated modifying the rejections to address the amendments. Applicant argued that Badescu does not disclose a cross-linker of Formula (I) as not required by claims 1, 3 and 14 as amended and thus even if a skilled person combined the teaching of Li and Badescu, they cannot arrive at the invention of Applicant’s claim. Applicant further argued that Badescu actually center on a cross-linker having an NHS structure as shown in Fig 2 of Badescu and this is entirely different structure to the Formula (I) of Applicant’s claimed invention. The above arguments have fully been considered but are not found persuasive. Applicant’s arguments that the linker of Badescu having an NHS structure is entirely different, does not have a clear basis because claim 1, in fact encompasses a linker that reads on Badescu’s linker for providing the conjugate. As for example, Badescu discloses trifunctional linker 4-[2,2-Bis[(p-tolylsulfonyl)methyl]acetyl) benzoic acid-NHS ester; bis-sulfone NHS ester (bi-sulfone NHS); i.e PNG media_image2.png 239 284 media_image2.png Greyscale ; page 1133, line 11 of 1st col) for providing conjugates with antibody. Badescu also teaches that the bis-reactive sulfone group (bis-alkylating group) is capable of undergoing reaction with both sulfur atoms derived from a reduced disulfide bond in antibodies and antibody fragments resulting in covalent re-bridging via a three-carbon bridge leaving the protein (antibody) structurally intact and the resulting conjugates are stable and retained antigen-binding (Abstract and introduction). Claim 1 is directed to a substrate onto which a biomolecule (see one or more biomolecule) are immobilized to the substrate surface by means of the linker of Formula (I) and the final structure of the conjugate with the linker is considered for patentability and not the linker by itself because not all part of the linker exist after attachment to the biomolecule and to the substrate. The linker of Formula (I) reads on the linker PNG media_image2.png 239 284 media_image2.png Greyscale wherein R1 is -phe-CH3, X is CH2, Y is -CH2-SO2-Ph-CH3 and the only difference is activated ester group NHS, which when reacts with a substrate having an amine group, would provide a linkage through NH and as for example, amine surface (surface with NH2) or surface with NH2 with a linker (EtO03Si(CH2)-NH-). Since Lin teaches immobilization of antibody on surface and Badescu teaches linker having bis-alkylating group for linking to thiols of antibody and NHS group for linking to other molecules through amine group, it would be obvious to one of ordinary skilled in the art to provide Lin with the linker of Badescu and various amine containing surfaces with the expectation of expanding the arsenal of surfaces for immobilization of antibody having retained antigen-binding ability with a reasonable expectation of success. Further, it is noted that Applicant’s arguments seems to be based on linker which is not been reacted with the biological molecule and the substrate but however, the claim is directed to a substate wherein the biological molecule is immobilized on the substrate through the linker and thus in the final product (which is claimed) the linker has already been reacted to the substrate and the biological molecules. Thus, in the final product, wherein the biological molecule is immobilized to the substrate through the linker, the linker will not retain the functional groups as described by R2 and R1. Conclusion THIS ACTION IS MADE FINAL. 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 SHAFIQUL HAQ whose telephone number is (571)272-6103. The examiner can normally be reached on Mon-Fri 8-4:30. Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice. If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Gregory S. Emch can be reached on 571-272-8149. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300. 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