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 .
Election/Restrictions
Applicant’s election without traverse of group I with species of protein A/G/L fusion protein in the reply filed on 11/25/2025 is acknowledged.
Claims 1, 5, 12, 15, 20-21, 23, 27- 28, and 30-31 read on the elected species of protein A/G/L are considered.
Claims 32-33, 36, 40-41, 43-44 are withdrawn from consideration.
Examiner’s Note:
(A) The Broadest Reasonable Interpretation (BRI) of crosslinking cited in the claim 20 is based on the well accepted concept or explanation by the state of art as evidenced by Therma Fisher SCIENTIFIC on line (2026):
Crosslinking is the process of chemically joining two or more molecules by a covalent bond. Modification involves attaching or cleaving chemical groups to alter the solubility or other properties of the original molecule. "Labeling" generally refers to any form of crosslinking or modification whose purpose is to attach a chemical group (e.g., a fluorescent molecule) to aid in detection of a molecule and is described with following technologies such as Protein and peptide biotinylation, Antibody labeling with fluorophores and biotin, Immobilizing biomolecules to surfaces and Capturing protein interaction.
Therefore, the antibody or protein labeling with a fluorescent or mobilizing the antibody and antibody-binding agent complete to the surface being detected or isolated directly or indirectly is considered as crosslinking.
(B) Claim 31 The Broadest Reasonable Interpretation (BRI) of the citation “ up to 5 days after dadding the aggregating to the agent to the biological sample : isa range from above 0 minute to 5 days.
Claim Rejections - 35 USC § 102
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.
Claim(s) 1, 15, 20, 23, 24, 27-28, 30 and 31 are rejected under 35 U.S.C. 102 (a) (1) as being anticipated by US Patent No. 9,442,112B2 to Mehra et al.
Mehra et al. describe a method at paragraph 72 for detecting an antibody in a sample, wherein the method comprises the following steps: a detectable specific binding partner for the antibody is added to a mixture comprising an antibody under the detection. Furthermore, the reaction mixture can be added with a detectable secondary antibody or other binding agent , wherein the other binding agent can be protein A, protein G , protein L or chimeric protein A, protein G, and protein L such as protein A/G or A/G/L, A/L, G/L or combinations thereof) which binds to the first antibody. This secondary antibody or other binding agent can be labeled, e.g., with a radioactive, enzymatic, fluorescent, luminescent, chemi-luminescent, metallic nanoparticle or metallic nanoshell (e.g. colloidal gold), or other detectable label, such as an avidin/biotin, avidin/streptavidin or avidin/polystreptavidin system. In another embodiment, the binding partner is a peptide of the invention, which can be conjugated directly or indirectly (e.g. via biotin/avidin or biotin/streptavidin interaction) to an enzyme, such as horseradish peroxidase or alkaline phosphatase or other signaling moiety. In such embodiments, the detectable signal is produced by adding a substrate of the enzyme that produces a detectable signal, such as a chromogenic, fluorogenic, or chemiluminescent substrate. Paragraphs 78-81 also described similar methods described above.
It is well known in the art that a fluorophore is defined in the state of art as fluorescent chemical compound that can re-emit light excitation. Fluorophores are sometimes used alone, as a tracer in fluids, as a dye for staining of certain structures, as a substrate of enzyme or as a probe or indicator (when its fluorescence is affected by environmental aspects such as polarity or ions). More generally they are covalently bonded to macromolecules, serving as a markers (or dyes, or tags, or reporters) for affine or bioactive reagents (antibodies, peptides, nucleic acids). Fluorophores are notably used to stain tissues, cells, or materials in a variety of analytical methods, such as fluorescent imaging and spectroscopy. Hence, the labeling agents such as fluorescent, luminescent of chemi-luminescent etc. cited prior art (See paragraph #4 cited supra) meet the limitaiton of claim 15.
Moreover, if the labeled agent is a radioactive material, the detectable signal can last up to 5 days, wherein if the labeling agent is fluorescent, luminescent, chemi-luminescent, the antibody-antibody binding agent aggregate is detected within 5 minutes of adding the aggregating agent to the biological sample. This is common science and technology well known in the art . Still further, the utilization of chemi-luminescent chemiluminometry labeling agent involves 470 nm blue light emission which inherently requires 470 nm LED Transilluminators for the detection of the target color for diagnostic tests, such as those measuring alkaline phosphatase levels. To this context the limitations cited in claims 27-28, 30 and 31 are met inherently or implicitly. Therefore, the cited reference anticipates claims 1, 15, 20, 23-24, 27-28, 30-31.
Claim Rejections - 35 USC § 102/103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
Claims 1, 5, 12, 15, 20, 21, 24, 27-28 and 30-31 are rejected under 35 U.S.C. 102 ( (a) (1) as anticipated by or, in the alternative, under 35 U.S.C. 103 as obvious over Burbelo et al. (Journal of Infectious Disease, May 19, 2020, Vol. 222, pages 206-213) further in view of Degeling et al. (Anal Chem. 2013 Feb 20;85(5):3006–3012) and Hawkins et al. (Promega Notes, 2005, Number 90, pages 1-14) for claim 27.
Burbebo et al. teach a luciferase immunoprecipitation system (LIPS) assay (LIPS). in which viral protein of SARS-Cov-2 nucleocapsid protein (NP) or spike protein (S) fused to a light-emitting luciferase protein to for Renilla luciferase fusion protein for NP and Gaussia luciferase for Spike protein respectively. Such antibody binding fusion protein complexes are used to bind and pulldown the antibody to NP or S respectively by an immunoprecipitation (IP) processes respectively. After incubations of the immunoprecipitations for 1 hour, each of the immunoprecipitation mixtures was transferred to a microtiter filter plate containing protein A/G beads and incubated for 1 hour. Protein A/G beads efficiently bind IgG and, to a much lesser extent, IgM, to capture the antibody-antigen-bead complexes. Following the IP, the capture the antibody-antigen-bead complexes were transferred to microtiter plates for watching to clean the unbound and uninterested proteins in microtiter plates 8 times with buffer A and twice with phosphate-buffered saline to remove unbound antigens. After the final wash, coelenterazine substrate (Promega) was added to detect the luciferase activity, and light units (LU) were measured using a Berthold LB 960 Centro microplate luminometer (Berthold Technologies), which is an instrument that is used for detection with a single wave length selected from the florescence emission range from 480 to 630 as evidenced by Berthold LB 960 Centro microplate luminometer (Berthold Technologies).
Furthermore, the cited reference also teaches all antibody binding agents of SARS2 NP- Gluc and SARS2 S-Rluc were expressed as recombinant fusion proteins or its fragment thereof are respectively produced by expressing them with expression vector pGAUS3 vector in a host cell line and used as an antibody binding agent separately to bind to different antibodies to SARS2 NP or SARS22 spike or its fragment thereof ( Please See Section of LIPS Assay for measurement of SARS-Cov-2 antibodies).
Regarding claim 27, the cited reference does not teach explicitly using the 470 nm wavelength to detect the blue light of Bioluminescence imaging (BLI) produced by either Gaussia luciferase (GLuc) protein and Renilla luciferase (Rluc) fusion with Berthold LB 960 Centro microplate luminometer. However, prior to the current Application was filed, an expert, Dr. Degeling et al. teach clearly that GLuc emits blue light with a maximum intensity typically around 470 nm that can reach 100% maximum in about 5 min (See Fig. 3 and the 2nd paragraph in the section of Introduction). Expert , Dr. Hawkins et al. clearly teach that the maximum intensity by Rluc can reach to 100% in 5 minutes at the wavelength 470 nm too (See Figs. 4 and 5 A). Hence, prior to the current Application was filed, it had been well known in the art , the wavelength for detecting the optimal intensity for both GLuc and Rluc fusion are both around 470 nm and detectable to the maximal level in 5 min.
Therefore, the cited reference anticipates claims 1, 5, 12, 15, 20, 24, 27, 28, 30-30 explicitly and implicitly.
Or alternatively, it would have been obviously for any person with an ordinarily skilled in the art to choose the optimal wavelength 470 nm as it is cited in claim 27 for detecting the Bioluminescence imaging (BLI) for both G Luc) and Rluc SARS-NP or SARS Spike protein by Berthold LB 960 Centro microplate luminometer, hereby for producing the most optimal results successfully.
Claim(s) 1, 5, 12, 15, 20, 23, 24, 27-28, 30-31 are rejected under 35 U.S.C. 102 (a) (1) as being anticipated by or in the alternative, under Haljasmägi et al. (European Journal102/ of Immunology, First published on line, 25, June 2020, https://doi.org/10.1002/eji.202048715.) further in view of Degeling et al. (Anal Chem. 2013 Feb 20;85(5):3006–3012).
Haljasmägi et al. disclose a sensitive method called Luciferase IP system (LIPS) assay for quantitative detection of antibodies to antigens in their native conformation, wherein the antibody is the one responses to SARS-CoV-2 spike (S) and one to nucleocapsid (N) proteins in COVID-19 patients. First, the SARS-CoV-2 antigens. S1, S2, and N genes were cloned in fusion with NanoLuc (Promega) gene and expressed in HEK293 cells. Which serves as the antibody binding agents with a detectable labeling of NanoLuc. The cell lysates of HEK293 rich in NanoLuc S1, S2 or N fusion proteins were incubated with plasma samples (in 1:40 dilution), which contain antibodies to be detected. The mixture of the antibody and antibody binding agent of NanoLuc S1, S2 or N fusion proteins were mixed with protein G Sepharose that is considered as aggregating agent, which allow the antibodies binding Protein G Sepharose to capture antibody/viral antigens complexes. After the washing, luciferase substrate Nano-Glo™ (Promega) was added and luminescence was measured in VICTOR X multilabel reader (PerkinElmer Life Sciences (See Abstract, the 1 and 2 paragraph of Introduction, Abstract, and Fig. 1 and its legend).
It is worth to note that the maximal luminescence for the Nano-Glo™ produced by HEK293 cells reach to the plateau at 5 hours as evidenced by Technical Manual of Promega in Nano-Glo® Luciferase Assay System, originally published in 2012-2015, pp 1-17, Please see (See Fig. 3 on page 5 ).
Still further regarding claim 27, the cited reference does not teach explicitly using the 470 nm wavelength to detect the blue light of Bioluminescence imaging (BLI) produced by NanoLuc, with Gaussia luciferase (GLuc) protein. However, prior to the current Application was filed, an expert, Dr. Degeling et al. teach clearly that GLuc emits blue light with a maximum intensity typically around 470 nm that can reach 100% maximum in about 5 min (See Fig. 3 and the 2nd paragraph in the section of Introduction). Hence, prior to the current Application was filed, it had been well known in the art , the wavelength for detecting the optimal intensity for both GLuc and Rluc fusion are both around 470 nm and detectable to the maximal level in 5 min.
Therefore, the cited reference anticipates claims 1, 5, 12, 15, 20, 24, 27, 28, 30-30 explicitly and implicitly.
Or alternatively, it would have been obviously for any person with an ordinarily skilled in the art to choose the optimal wavelength 470 nm as it is cited in claim 27 for detecting the Bioluminescence imaging (BLI) for G Luc) and Rluc SARS-NP or SARS Spike protein with the optimal wavelength at 470 nm utilization hereby for producing the most optimal results successfully.
MPEO 2105 cites: III. A REJECTION UNDER 35 U.S.C. 102/103 CAN BE MADE WHEN THE PRIOR ART PRODUCT SEEMS TO BE IDENTICAL EXCEPT THAT THE PRIOR ART IS SILENT AS TO AN INHERENT CHARACTERISTIC. Where applicant claims a composition in terms of a function, property or characteristic and the composition of the prior art is the same as that of the claim but the function is not explicitly disclosed by the reference, the examiner may make a rejection under both 35 U.S.C. 102 and 103, expressed as a 102/103 rejection. “There is nothing inconsistent in concurrent rejections for obviousness under 35 U.S.C. 103 and for anticipation under 35 U.S.C. 102.” In re Best, 562 F.2d 1252, 1255 n.4, 195 USPQ 430, 433 n.4 (CCPA 1977). This same rationale should also apply to product, apparatus, and process claims claimed in terms of function, property or characteristic. Therefore, a 35 U.S.C. 102/103 rejection is appropriate for these types of claims as well as for composition claims.
Claims 1, 15, 20, 21, 27-28 and 30-31 are rejected under 35 U.S.C. 102 (a) (1) as being anticipated or in an alternative obviously by Arai et al. (JOURNAL OF FOERMATION AND BIOENGNEERING , 1998, vol. 86, No. 5, pp. 440-445) for claims 1, 15, 20-21, 28, 31-32 with a further in view of Kain et al. (Expression and detection of Green Fluorescent Protein (GFP) published in 1997, Vol. 63, pp. 305-324) for claim 27.
Arai et al. teach a method for detecting an antibody in a biological sample comprising contacting the biological sample comprising an antibody as either goat IgG, rabbit IgG, sheep IgG and mouse IgG1 with an antibody binding agent is an antigen to each of the antibody , such as goat antibody and binding to the secondary antibody to the antigen and its conjugating labeling agent associated with an aggregating agent comprising protein G (PG) and a fluorophore agent i.e. fluorescent-enhancing green fluorescent protein (EGFP), wherein the antibody binding agent is conjugated with flowing the complex formation cited above,
In particular, the Fluoroimmunoassays comprising the following steps in general : 1). Generation of the recombinant PG-EGFP or EGFP fusion protein aggregating complex by recombinant DNA technology from host cell E. Coli JM 109 and purified ready to use for the process of an antibody-antibody binding agent complex formation (Fig. 1). The fluorophore emission produced by PG-EGFP was measured by a fluorescence spectra using fluorescence spectrophotometer F-2000 with emission excitation wavelength at 488 nm and excitation spectra by emission at 509 nm. Prior ot the complex formation, each kind of antibody was coated into wells, blocking unnecessary background and adding antigen and binding antigen to the coating antibody=, binding secondary antibody to the antigen and conjugating PE-EGFP aggregation to the secondary antibody. The maximal emission analysis for PG-EGF or FITC last more than 10 min around 500 nm intensity shown in Fig. 3 by the spectra measurement between 488 nm to 509 nm. It is clearly to see that with GP conjugation to the IgG Fc , GFP has better emission excitation in intensity (See Fig. 4) within 10 min observation length. (See Figs. 1-4 and sections of Materials and Methods on page 440-445 for detail)
Therefore, the cited references teach the limitations of claims 1, 15, 20,21, 28, and 30 and 31 explicitly, while the cited reference dis not explitely teach using the 470 nm for the blue light LED detection explicitly.
An expert of fluorophore emission prior to the current Application by Kain et al. teach that “The wt GFP absorbs UV and blue light with a maximum absorbance at 395 nm and a minor peak of absorbance at 470 nm. The GFP-S65T and RSGFP variants have a single excitation peak at approx. 490 nm (Fig. 3). The shifted spectra of the variant proteins has advantages for both fluorescence microscopy and FACS analysis. The filter sets commonly used with fluorescein isothiocyanate (FITC) and GFP illuminate at 450–500 nm—well above the major excitation peak of wt GFP. These filter sets therefore produce a fluorescent signal from wt GFP that is several-fold less than would be obtained with filter sets that excite at 395 nm. However, use of the major peak at 395 nm for excitation of the wt GFP is not advisable owing to rapid photobleaching of the fluorescent signal. For excitation of the wt GFP, we recommend the 470 nm peak to minimize photobleaching (See section 3.2)”.
Hence, alternatively, it would have been obviously for any person with an ordinarily skilled in the art to choose the optimal wavelength 470 nm drafted in claim 27 for detecting the Bioluminescence imaging (BLI) for both G Luc) and Rluc SARS-NP or SARS Spike protein by Berthold LB 960 Centro microplate luminometer, hereby for producing the most optimal results successfully.
MPEO 2105 cites: III. A REJECTION UNDER 35 U.S.C. 102/103 CAN BE MADE WHEN THE PRIOR ART PRODUCT SEEMS TO BE IDENTICAL EXCEPT THAT THE PRIOR ART IS SILENT AS TO AN INHERENT CHARACTERISTIC. Where applicant claims a composition in terms of a function, property or characteristic and the composition of the prior art is the same as that of the claim but the function is not explicitly disclosed by the reference, the examiner may make a rejection under both 35 U.S.C. 102 and 103, expressed as a 102/103 rejection. “There is nothing inconsistent in concurrent rejections for obviousness under 35 U.S.C. 103 and for anticipation under 35 U.S.C. 102.” In re Best, 562 F.2d 1252, 1255 n.4, 195 USPQ 430, 433 n.4 (CCPA 1977). This same rationale should also apply to product, apparatus, and process claims claimed in terms of function, property or characteristic. Therefore, a 35 U.S.C. 102/103 rejection is appropriate for these types of claims as well as for composition claims.
Conclusion
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BAO Q. LI
Examiner
Art Unit 1671
/BAO Q LI/Primary Examiner, Art Unit 1671