DETAILED ACTION
Non-Final Rejection
Notice of Pre-AIA or AIA Status
1. 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
2. Applicant’s election of Group I, claims 1-6, 8-9, 12 and 14-15 with species election “mammalian cell” in the reply filed on 02/13/2026 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)).
The Restriction/Election is made final.
Status of Claims
3. Claims 1-6, 8-9, 12, 14-16, 18, 23, 25-27, 29 and 31-32 are pending.
4. Claims 16,18,23,25-27,29 and 31-32 withdrawn due to Restriction/Election.
5. Claims 1-6, 8-9, 12 and 14-15 are under examination.
Priority
6. This 18/275,226 application is 371 of PCT/CN2022/077527 filed on 02/23/2022 and claims an earlier priority to date of filing date 03/31/2021 to CN202110348881.6 filed in country People’s Republic of China.
Information Disclosure Statement
7. The information disclosure statement (IDS) submitted on 07/05/2024 is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Specification Objections
8. The disclosure is objected to because it contains an embedded hyperlink and/or other form of browser-executable code. Applicant is required to delete the embedded hyperlink and/or other form of browser-executable code; references to websites should be limited to the top-level domain name without any prefix such as http:// or other browser-executable code. See MPEP § 608.01.
The instant specification para [0022] has the embedded hyperlink: (https:/iwwwncbi.nlm,nih.gov/).
9. This application contains sequence disclosures that are encompassed by the definitions for nucleotide and/or amino acid sequences set forth in 37 CFR 1.821(a)(1) and (a)(2). However, this application fails to comply with the requirements of 37 CFR 1.821 through 1.825 for the reason(s) set forth below or on the attached Notice To Comply With Requirements For Patent Applications Containing Nucleotide Sequence And/Or Amino Acid Sequence Disclosures.
The specification is objected to because there is no incorporation by reference statement of the Sequence Listing. This statement is required. Applicants must comply with sequence rules in order to be considered a complete response to this Office Action.
10. The lengthy specification has not been checked to the extent necessary to determine the presence of all possible minor errors. Applicant’s cooperation is requested in correcting any errors of which applicant may become aware in the specification.
Claim Objections
11. The claims filed in claim listing on 08/01/2023 are objected to because the lines are crowded too closely together, making reading difficult. Substitute claims with lines one and one-half or double spaced on good quality paper are required. See 37 CFR 1.52(b).
12. In the claim 1, in the absence of a reference sequence (SEQ ID NO) it is confusing to interpret to which reference sequence numbering the claimed the amino acid sequence positions 319-537 in an RBD domain of an S protein relates to.
13. Claims 3 and 8 are also objected to because: the instant claim 3 should recite “SEQ ID NO: 1”, and the instant claim 8 should recite “SEQ ID NO: 2”, respectively, instead of “SEQ ID NO. 1” and “SEQ ID NO. 2”, respectively. The period used need to be replaced with colon.
Applicants must comply with sequence rules in order to be considered a complete response to this office action.
Claim Rejections - 35 USC § 112
14. 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 1-6, 8-9, 12 and 14-15 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim 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.
The claims 1-4, and 6 recites “novel coronavirus”. The term “novel” is not a taxonomical term allowing to identify the virus referred to. It is not clear to which coronavirus the prefix “novel” refers to, and therefore the claims 1-6, 8-9, 12 and 14-15 are indefinite for failing to particularly point out and distinctly claim the subject matter.
Claim Interpretation
15. The claims in this application are given their broadest reasonable interpretation using the plain meaning of the claim language in light of the specification as it would be understood by one of ordinary skill in the art.
Claim 1: The instant claim 1 is directed to an S-RBD trimer protein for a novel coronavirus, the trimer protein composed of amino acid fragments at positions 319-537 in an RBD domain of an S protein of the novel coronavirus in a trimer form.
“A novel coronavirus” in light of specification para [0078] is interpreted to refer to SARS-CoV-2 virus. The S-RBD trimer is interpreted to be formed by SARS-CoV-2 S protein fragment amino acid sequence positions 319-537 in an RBD domain without introducing exogenous sequence or vector sequence (instant specification, Example 1 para [0087]).
Claim 12: “A host cell” is interpreted as a cell line or CHO cell line (instant specification, para [0083], [0089]-[0090]).
Claim Rejections - 35 USC § 101
16. 35 U.S.C. 101 reads as follows:
Whoever invents or discovers any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements of this title.
17. Section 33(a) of the America Invents Act reads as follows:
Notwithstanding any other provision of law, no patent may issue on a claim directed to or encompassing a human organism.
Claims 12, and 14-15 are rejected under 35 U.S.C. 101 and section 33(a) of the America Invents Act as being directed to or encompassing a human organism. See also Animals - Patentability, 1077 Off. Gaz. Pat. Office 24 (April 21, 1987) (indicating that human organisms are excluded from the scope of patentable subject matter under 35 U.S.C. 101).
The instant claims 12, and 14-15 recite the limitation of “a host cell”, which reads upon a human organism and human organisms are not patentable subject matter. It is suggested that the claims be amended to recite “an isolated host cell”, which would not read upon a human organism.
Claim Rejections - 35 USC § 103
18. 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 (i.e., changing from AIA to pre-AIA ) 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.
19. Claims 1-2 are rejected under 35 U.S.C. 103 as being unpatentable over Dai et al 2020 (Cell. 2020 Aug 6;182(3):722-733.e11), and further in view of Yang et al 2020 (Nature, 586(7830), 572-577), Webb et al 2017 (Curr Protoc Bioinformatics; 54: 5.6.1–5.6.37), and Cai et al 2020 (Science, 369(6511), 1586-1592).
Claims 1-2: Dai et al 2020 is the art and teaches a SARS-CoV-2 RBD dimer comprising two units of RBD amino acids R319-K537 fragment dimerized as tandem repeat linking two of the amino acid fragments in an order from an N terminal to a C terminal (See, page 727, Fig 3-A). Dai et al 2020 teaches full-length trimeric S protein is usually highly immunogenic due likely to its large size (See, page 728, col 2, para 3, entire article).
Dai et al 2020 does not teach a trimer protein comprising RBD amino acids R319-K537.
Yang et al 2020 is in the art and teaches a recombinant vaccine that comprises residues 319–545 of the RBD of the spike protein induces a potent functional antibody response in immunized mice, rabbits and non-human primates as early as 7 or 14 days after the injection of a single vaccine dose (See, abstract, entire article).
Webb et al 2017 is in the protein modeling art and teaches comparative protein structure modeling using MODELLER to predicts the three-dimensional structure of a given protein sequence (target) based primarily on its alignment to one or more proteins of known structure (templates) (See, abstract, entire article, and figures).
Cai et al 2020 is in the art and teaches distinct conformational states of SARS-CoV-2
spike protein and RBD protein (See, abstract, Fig. 5, entire article).
It would have been obvious to one of the ordinary skills in the art before the effective filing date of the claimed invention to modify the prior art teachings of Dai et al 2020 on RBD dimer of SARS-CoV-2, and trimeric nature of spike protein of SARS-CoV-2 virus with additional teachings of Yang et al 2020 on RBD comprising amino acid residues 319–545 induced a potent functional antibody response, Webb et al 2017 on comparative protein structure modeling, and Cai et al 2020 on confirmational states of S protein to arrive at the inventions of claims 1-2. One of the ordinary skills would have been motivated to increase the immunogenicity of the claimed RBD trimer comprising three amino acid 319-537 fragments to develop an efficacious and safer vaccine against SARS-CoV-2 virus and for commercial success. There would be a reasonable expectation of success given the applied prior art teachings in the art as recited supra. This is analogous to some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the invention as claimed in claims 1-2. See KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007) (see MPEP § 2143, example of rationales, A -G).
20. Claim 3 is rejected under 35 U.S.C. 103 as being unpatentable over combined teachings of Dai et al 2020 (Cell. 2020 Aug 6;182(3):722-733.e11), Yang et al 2020 (Nature, 586(7830), 572-577), Webb et al 2017 (Curr Protoc Bioinformatics; 54: 5.6.1–5.6.37), and Cai et al 2020 (Science, 369(6511), 1586-1592) as applied to claims 1-2 above and further in view of Dai et al 2020 (CN111592602A, 08/28/2020), and Georges et al 2021 (WO2021163536A2, 08/19/2021, with an earlier priority to US provisional 62/977,078 filed on 02/14/2020).
Claim 3: The combined teachings of Dai et al 2020, Yang et al 2020, Webb et al 2017, and Cai et al 2020 teaches claims 1-2 as recited supra. However, does not teach the SEQ ID NO: 1 of claim 3.
Dai et al 2020 further disclosed SARS-CoV-2 spike protein amino acid sequence (See, page e6, SARS-CoV-2 (GenBank: QHR63290) Sequences used in the alignments (NCBI GenBank Accession number QHR63290.2) as recited below (underlined amino acid sequence Qy is instant SEQ ID NO: 1; Db is amino acid sequence in QHR63290.2, see pdf printout).
The instant SEQ ID NO: 1 consists of a methionine at position 1 that is required for initiation of translation followed by three repeats of the R319-K357 portion of RBD (See, Dai et al 2020, Figure 3A) that discloses two copies of RBD (R319-K537) fused in tandem.
Qy 200 -----LLHAPATVCGPK-----KSTNLVKNKRVQPTESIVRFPNITNLCPFGEVFNATRF 249
| | | | | | |||||||||||||||||||||||||||||
Db 288 AVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRF 347
Qy 250 ASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEV 309
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 348 ASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEV 407
Qy 310 RQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERD 369
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 408 RQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERD 467
Qy 370 ISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGP 429
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 468 ISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGP 527
Qy 430 KKSTNLVKNKRVQ------------------------------PTESIVRFPNITNL--- 456
|||||||||| | | || | :
Db 528 KKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDI 587
Additionally, Dai et al 2020 (CN111592602A, 08/28/2020) also discloses essentially the same subject-matter as that of an article by Dai et al 2020 (Cell, 2020). The SEQ ID NO: 6 of (Dai et al 2020, CN111592602A) is 100% identical with present SEQ ID NO: 1 in a 438 amino acid overlap). The difference lies in the third fused repeat in the instant SEQ ID NO: 1. Dai et al 2020 research article recited that trimeric S protein constructs elicit higher immune response. Therefore, consequently, the effect disclosed (instant specification, e.g., Fig 14-15, para [00109] in Table 3), increased antibody titer of the RBD trimer over the RBD dimer, was to be expected. Furthermore, even if the exemplified effects of the instant SEQ ID NO: 1 RBD trimer would be considered non-obvious, it would not be plausible that a 95% homologous amino acid sequence would retain the same effects as that of the 100% identical amino acid sequence.
Georges et al 2021 is in the SARS-CoV-2 virus vaccine art and teaches SEQ ID NO: 475 (Db) that has 97.6% identity to the instant SEQ ID NO: 1 (Qy) as recited below, and therefore render obvious the added limitation of instant claim 3 a sequence having a homology of 95% with SEQ ID No. 1. It is obvious to include sequence to encode Methionine (M) at position 1 for initiation of translation.
FEATURE: OTHER INFORMATION: SARS-CoV-2 virus spike protein
Query Match 97.8%; Score 3484; Length 1527;
Best Local Similarity 89.2%;
Matches 654; Conservative 1; Mismatches 2; Indels 76; Gaps 2;
Qy 2 RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFK 61
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 18 RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFK 77
Qy 62 CYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNS 121
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 78 CYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNS 137
Qy 122 NNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQ 181
||||||||||||| ||||||||||||||||||||||||||||||||||||||||||||||
Db 138 NNLDSKVGGNYNYRYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQ 197
Qy 182 PTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNK--------------------- 220
|||||||||||||||||||||||||||||||||||||||
Db 198 PTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKSVNFTFGGGGSGGGGSGGGGS 257
Qy 221 -----------------RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNC 263
|||||||||||||||||||||||||||||||||||||||||||
Db 258 TLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNC 317
Qy 264 VADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADY 323
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 318 VADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADY 377
Qy 324 NYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCN 383
|||||||||||||||||||||||||||||| |||||||||||||||||||||||||||||
Db 378 NYKLPDDFTGCVIAWNSNNLDSKVGGNYNYRYRLFRKSNLKPFERDISTEIYQAGSTPCN 437
Qy 384 GVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNK---- 439
||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 438 GVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKSVNF 497
Qy 440 ----------------------------------RVQPTESIVRFPNITNLCPFGEVFNA 465
||||||||||||||||||||||||||
Db 498 TFGGGGSGGGGSGGGGSTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNA 557
Qy 466 TRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRG 525
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 558 TRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRG 617
Qy 526 DEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF 585
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 618 DEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF 677
Qy 586 ERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATV 645
|||||||||||||||||||:||||||||||||||||||||||||||||||||||||||||
Db 678 ERDISTEIYQAGSTPCNGVKGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATV 737
Qy 646 CGPKKSTNLVKNK 658
|||||||||||||
Db 738 CGPKKSTNLVKNK 750
Therefore, the combined teachings as applied to claim 3 renders obvious the added limitation of instant claim 3, wherein the S-RBD trimer protein for the SARS-CoV-2 virus comprises an amino acid sequence shown as SEQ ID No. 1 or a sequence having a homology of 95% or higher with SEQ ID NO: 1. One of the ordinary skills would have been motivated to develop the optimal RBD trimer construct using the known available sequence for developing highly immunogenic vaccine for commercial purpose with a reasonable expectation of success based on the prior art knowledge and the ordinary skills in the art.
21. Claims 4-6, 8-9, 12, 14, and 15 are rejected under 35 U.S.C. 103 as being unpatentable over combined teachings of Dai et al 2020 (Cell. 2020 Aug 6;182(3):722-733.e11), Yang et al 2020 (Nature, 586(7830), 572-577), Webb et al 2017 (Curr Protoc Bioinformatics; 54: 5.6.1–5.6.37), and Cai et al 2020 (Science, 369(6511), 1586-1592) as applied to claims 1-2 above and further in view of Yang et al 2020 (CN111944064A, 11/17/2020), Gong et al 2020 (CN111533809A, 08/14/2020), and Fritz et al 2022 (WO2022006357A2, 01/06/2022, with an earlier priority to US provisional 63/046426 of 06/30/2020), and Inouye et al 2015 (Protein Expression and Purification, 109, 47-54)
Claims 4-5: The combined teachings of Dai et al 2020, Yang et al 2020, Webb et al 2017, and Cai et al 2020 teaches claims 1-2 as recited supra. However, does not teach the added limitations of claims 4 and 5.
Yang et al 2020 (CN111944064A) is in the art and teaches an added limitation of instant claims 4 a fusion protein (instant claim 4), and a signal peptide (instant claim 5) by disclosing a human interleukin 10 (IL-10) signal peptide, a sequence from N-terminus to C-terminus: human interleukin 10 signal peptide, S-S-RBD, foldon protein (See, claim 1). In the art it is know that the IL-10 signal peptide sequence is responsible for directing the protein to the secretory pathway and is cleaved to produce the mature protein, in the instant claim e.g. RBD trimer.
Gong et al 2020 (CN111533809A) is in the art and teaches instant claim 4 and 5 added limitations by disclosing the fusion protein (SARS-CoV-2-RBD-Fc) by fusing the RBD structure domain of the S protein of SARS-CoV-2 virus and the antibody Fc fragment, and the fusion protein can be used as subunit vaccine, and can induce subject to generate high-efficiency neutralizing antibody (Fc as an immune-enhancing peptide) by nasal drip immunization and intramuscular injection, which shows that SARS-CoV-2-RBD-Fc can be used as candidate vaccine for preventing disease and treating new coronavirus infection (See, abstract, claim 1).
Claims 6, 8-9, 12, 14, and 15: The combined teachings of Dai et al 2020, Yang et al 2020, Webb et al 2017, and Cai et al 2020 teaches claims 1-2 that comprise S-RBD trimer protein of SARS-CoV-2 virus. Therefore, a nucleic acid molecule, comprising a nucleotide sequence encoding the S-RBD trimer protein for the SARS-CoV-2 virus according to claim 1 is rendered obvious by the applied prior art as recited below.
Dai et al 2020 as recited supra teaches design of RBD dimer protein (Fig 3, A) and the RBD 319-537 protein fragment amino acid sequences as taught for claim 3 above (See, above amino acid sequence alignment), RBD protein expression is taught in mammalian cells (See, page e6) by disclosing the coding sequence for RBD protein of SARS-CoV-2 were codon-optimized for mammalian cell expression in a construct for clinical grade CHO-K1 (Chinese hamster ovary ) cell line to express RBD. A strategy for codon-optimized MERS-CoV RBD dimer (reads on RBD of SARS-CoV-2) for cloning into the baculovirus transfer vector pFastBac1, insect cells, expression is disclosed (See, page e4- Methods). The HEK293T mammalian cell-expressed recombinant RBD proteins is disclosed with codon optimized sequences, these constructs were cloned into the pCAGGS vector, respectively, and transiently transfected into HEK293T cells (See, page e4- Methods).
Claim 8: Fritz et al 2022 is in art of SARS-CoV-2 recombinant polypeptide containing at least one immunogenic spike glycoprotein and an antibody Fc region, and has disclosed SEQ ID NO: 164 that has 76% identity match with instant SEQ ID NO: 2 of instant claim 8 as shown below.
Query Match 40.4%; Score 798.2; Length 2019;
Best Local Similarity 76.4%;
Matches 980; Conservative 0; Mismatches 303; Indels 0; Gaps 0;
Qy 38 CCAATATCACTAACCTGTGCCCCTTTGGGGAGGTGTTCAATGCCACAAGATTTGCTTCTG 97
|||||||||| ||| ||||||| ||||||||||| ||||||||||| | ||||| || |
Db 17 CCAATATCACAAACTTGTGCCCATTTGGGGAGGTCTTCAATGCCACTCGGTTTGCATCAG 76
Qy 98 TGTATGCCTGGAATAGAAAGAGAATCAGTAACTGTGTTGCTGACTACTCTGTGCTGTACA 157
| ||||| |||||| | || | || || || ||||||||||| || || || || |
Db 77 TATATGCATGGAATCGTAAACGGATATCAAATTGCGTTGCTGACTATTCAGTCCTCTATA 136
Qy 158 ATTCTGCCTCTTTCAGCACATTCAAATGTTATGGGGTTAGCCCCACCAAACTAAATGACC 217
|| ||| |||||| ||||| ||||| || || || || || || | |||||||
Db 137 ATAGTGCTAGTTTCAGTACATTTAAATGCTACGGAGTATCTCCTACTAAGTTGAATGACC 196
Qy 218 TGTGCTTCACCAATGTGTATGCTGACAGCTTTGTGATCAGAGGTGATGAAGTGAGACAAA 277
| |||||||| ||||| ||||||||||| ||||| || || ||||| || ||| | || |
Db 197 TTTGCTTCACAAATGTCTATGCTGACAGTTTTGTTATTAGGGGTGACGAGGTGCGGCAGA 256
Qy 278 TTGCCCCTGGGCAGACTGGCAAGATAGCTGACTACAATTACAAGCTGCCTGATGACTTTA 337
|||| || ||||| || || ||||| ||||||||||||||||| || ||||||||||| |
Db 257 TTGCACCCGGGCAAACAGGAAAGATCGCTGACTACAATTACAAACTTCCTGATGACTTCA 316
Qy 338 CTGGCTGTGTGATTGCCTGGAATAGCAATAACCTGGACAGCAAGGTGGGGGGCAACTATA 397
|||| ||||||||||| |||||| ||| || ||||| ||| ||||| || || || |
Db 317 CTGGTTGTGTGATTGCTTGGAATTCCAACAATCTGGATTCCAAAGTGGGTGGGAATTACA 376
Qy 398 ACTACCTATACAGATTGTTTAGAAAGTCCAATTTGAAACCCTTTGAGAGAGATATCAGCA 457
| ||| | || |||||||||||||| || || | || || || || | ||||| ||||
Db 377 ATTACTTGTATAGATTGTTTAGAAAATCTAACCTCAAGCCATTCGAACGGGATATAAGCA 436
Qy 458 CTGAGATCTATCAAGCTGGCAGCACCCCCTGTAATGGGGTGGAGGGGTTCAACTGCTATT 517
| || |||||||| ||||| ||||| || ||||| || || ||||||||||| |||||||
Db 437 CAGAAATCTATCAGGCTGGGAGCACACCTTGTAACGGCGTAGAGGGGTTCAATTGCTATT 496
Qy 518 TCCCCTTACAGAGCTATGGCTTTCAGCCCACCAATGGGGTGGGCTATCAGCCCTACAGAG 577
| || | ||| || |||||||| ||||||||||||||||| || ||||| || |||
Db 497 TTCCACTTCAGTCATACGGCTTTCAACCCACCAATGGGGTGGGATACCAGCCATATCGAG 556
Qy 578 TGGTGGTGCTGTCTTTTGAACTACTTCATGCCCCTGCCACAGTGTGTGGCCCCAAAAAAA 637
|||| || || ||||| || || ||||||||||| || || |||||||| ||||||||||
Db 557 TGGTCGTACTCTCTTTCGAGCTGCTTCATGCCCCCGCAACCGTGTGTGGTCCCAAAAAAA 616
Qy 638 GCACAAACCTGGTGAAAAACAAGAGAGTGCAGCCAACAGAGAGCATTGTGAGATTCCCTA 697
|||| || || || || || ||| || | | | || | |
Db 617 GCACCAATCTCGTCAAGAATAAGGGATCAGGGGGGGGCGGATCAGGCGGTGGAGGCAGCA 676
Qy 698 ACATAACCAATCTGTGTCCCTTTGGTGAGGTTTTTAATGCTACTAGATTTGCCTCTGTGT 757
||||||| ||||| ||||| || ||||| || |||||||| || | ||||| |||||||
Db 677 ACATAACTAATCTTTGTCCATTCGGTGAAGTCTTTAATGCAACCCGTTTTGCTTCTGTGT 736
Qy 758 ATGCCTGGAACAGAAAAAGAATCTCAAACTGTGTGGCTGACTATTCAGTCCTCTACAACT 817
|||| |||||||| |||||||| || || || || || |||||| ||||| ||||| |
Db 737 ATGCATGGAACAGGAAAAGAATATCCAATTGCGTTGCCGACTATAGCGTCCTTTACAATT 796
Qy 818 CAGCTAGCTTTAGCACCTTCAAATGCTATGGTGTCAGTCCCACCAAACTGAATGACCTTT 877
| || ||||||||| || || || ||||| ||| ||||| || |||||||| || |
Db 797 CTGCCTCCTTTAGCACATTTAAGTGTTATGGGGTCTCCCCCACTAAGCTGAATGATCTGT 856
Qy 878 GTTTCACCAATGTCTATGCTGACAGCTTTGTGATCAGAGGGGATGAAGTGAGACAAATAG 937
|||||||||| || ||||| ||| |||||| || | ||||||||||| |||||||||
Db 857 GTTTCACCAACGTTTATGCAGACTCCTTTGTTATACGGGGGGATGAAGTTCGACAAATAG 916
Qy 938 CCCCTGGGCAGACTGGCAAGATAGCTGATTACAACTATAAACTGCCTGATGATTTCACTG 997
| || |||||||| || ||||| ||||| ||||| || || || || ||||| |||||||
Db 917 CACCCGGGCAGACAGGGAAGATCGCTGACTACAATTACAAGCTTCCAGATGACTTCACTG 976
Qy 998 GCTGTGTCATTGCCTGGAACTCAAATAATCTAGACAGCAAGGTAGGTGGCAATTATAACT 1057
| || || || ||||||||| || || | ||| || || || || |||||||| |
Db 977 GGTGCGTAATCGCCTGGAACAGTAACAACTTGGACTCTAAAGTCGGAGGGAATTATAATT 1036
Qy 1058 ACCTCTATAGGCTGTTTAGGAAAAGTAACCTGAAACCCTTTGAGAGAGACATTAGCACAG 1117
|||| ||| | || || | ||||||||| |||| || |||||||| |||||| |||||
Db 1037 ACCTTTATCGTCTCTTCCGCAAAAGTAACTTGAAGCCTTTTGAGAGGGACATTTCCACAG 1096
Qy 1118 AGATTTATCAAGCTGGCTCAACACCCTGTAATGGTGTGGAGGGCTTTAACTGCTACTTCC 1177
|||| |||||||| || || ||||| || || || || ||||| ||||| || || ||||
Db 1097 AGATATATCAAGCAGGTTCCACACCATGCAACGGCGTCGAGGGATTTAATTGTTATTTCC 1156
Qy 1178 CCCTGCAGAGCTATGGCTTTCAGCCAACCAATGGAGTGGGCTATCAGCCCTATAGAGTAG 1237
|||| || || |||||||||||||| || || || ||||| || || || ||||| |
Db 1157 CCCTCCAATCATACGGCTTTCAGCCAACTAACGGCGTTGGCTACCAACCATACAGAGTCG 1216
Qy 1238 TTGTGCTGAGCTTTGAGCTGCTGCATGCCCCTGCCACAGTGTGTGGCCCTAAGAAAAGCA 1297
| ||||| |||||| |||||||| ||||||||||| ||||||||||| || || ||||
Db 1217 TAGTGCTTTCCTTTGAACTGCTGCACGCCCCTGCCACTGTGTGTGGCCCAAAAAAGAGCA 1276
Qy 1298 CCAACTTGGTCAAAAATAAGAGA 1320
|||| ||||| || || ||| ||
Db 1277 CCAATTTGGTAAAGAACAAGGGA 1299
Fritz et al 2022 as recited supra does not teach nucleic acid molecule comprises the nucleotide sequence shown as SEQ ID NO: 2 or a sequence having a homology of 95% or higher with SEQ ID NO: 2. According to instant specification (See, para [0020]) SEQID NO:2 is a codon optimized nucleotide sequence for the RBD trimer protein. Codon optimization is not novel and is disclosed by Dai et al 2020 as recited supra.
The prior art teachings of Dai et al 2020 as recited supra (also see below amino acid alignment) teaches 100% identity of amino acid sequence identity to the claimed RBD protein single unit of the trimer (claimed are three units of identical of RBD sequence placed in tandem), and thus the nucleotide sequence encoding the monomer of RBD with claimed amino acid positions 319-537 is known by prior art Dai et al 2020. Similarly, Fritz et al 2022 teaches RBD monomer amino acid positions 319-537 encoding nucleotide sequence.
Inouye et al 2015 teaches codon optimization of genes for efficient protein expression in mammalian cells by selection of only preferred human codons (See, entire article).
Below is shown partial instant SEQ ID NO: 1 RBD amino acid sequence for monomer of RBD with claimed amino acid positions 319-537 (Qy). The SEQ ID NO: 1 was trimmed for alignment.
Dai et al 2020 disclosed SARS-CoV-2 spike protein amino acid sequence (See, page e6, SARS-CoV-2 (GenBank: QHR63290) Sequences used in the alignments (NCBI GenBank Accession number QHR63290.2) as RBD 319-537. The amino acid sequence GenBank ID: QHR63290.2 was trimmed (Db) for alignment as shown below.
Query Match 100.0%; Score 1186; DB 1; Length 219;
Best Local Similarity 100.0%;
Matches 219; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFK 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFK 60
Qy 61 CYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNS 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 CYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNS 120
Qy 121 NNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQ 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 NNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQ 180
Qy 181 PTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNK 219
|||||||||||||||||||||||||||||||||||||||
Db 181 PTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNK 219
Dai et al 2020 as recited supra in the office action has expressed dimer of RBD protein fragment with 319-537 amino acid sequences positions in a mammalian cell, CHO cell line. Therefore, because the combined prior art teachings as recited supra has rendered obvious the claimed RBD trimer protein comprising RBD 319-537 protein fragment amino acid sequences, absent any evidence to the contrary, the same combined prior art teaching has rendered obvious the added limitation of instant claims 6 and 8, wherein the nucleic acid molecule comprises codon optimized nucleotide sequence similar to a nucleotide sequence having a homology of 95% or higher with SEQ ID No: 2 that encode the claimed S-RBD trimer protein of claim 1 . The search for the claimed SEQ ID NO: 2 sequence did not yield a sequence having a homology of 95% or higher with SEQ ID NO: 2 because (i) the SEQ ID NO: 2 is a nucleotide sequence that encode the repeat sequence RBD 319-537 amino acid coding nucleotide sequence in tandem (to construct a trimeric sequence) that does not occur in nature, (ii) the claimed SEQ ID NO: 2 nucleic acid sequence is a codon optimized sequence (See, instant specification para [0020]). The codon optimization for a desired species (e.g. mammalian, insect, yeast, E. coli) is known in the art. The prior art Dai et al 2020 as recited supra disclose codon optimization of nucleic acid sequence for increased expression in mammalian or insect cells. Therefore, absent any evidence to the contrary (e.g. higher RBD 319-537 protein expression than disclosed in the applied prior art), the combined prior art teaches instant claims 6 and 8 nucleic acid sequence SEQ ID NO: 2 that encode the RBD protein trimer of claim 1. In addition, Fritz et al 2022 discloses the nucleotide sequence of spike protein RBD and Inouye et al 2015 on codon optimization of nucleotide sequence for expression in mammalian cells, absent any evidence to the contrary, the combined prior art teachings rendered obvious instant SEQ ID NO: 2 nucleotide sequence that encode SARS-CoV-2 virus S-RBD trimer of claim 1 although there is no 95% or higher identity to the claimed SEQ ID NO: 2.
Therefore, it would have been obvious to one of the ordinary skills in the art before the effective filing date of the claimed invention to modify the combined prior art teachings applied to claims 1-2 with additional teachings of Dai et al 2020 on codon optimization and RBD sequence, Yang et al 2020, Webb et al 2017, Cai et al 2020, Yang et al 2020 (CN111944064A), Gong et al 2020 (CN111533809A) on mammalian cell, CHO cell expression of the trimeric RBD protein, Fritz et al 2022 on the nucleotide sequence of spike protein RBD and Inouye et al 2015 on codon optimization of nucleotide sequence for expression in mammalian cells as recited supra to arrive at the claims 6, 8-9, 12, 14, and 15. One of the ordinary skills would have been motivated to increase the immunogenicity of the claimed RBD trimer comprising three tandem repeats of RBD protein 319-537 amino acid fragments and to produce the protein in high quantity in mammalian cell to develop an efficacious and safer vaccine against SARS-CoV-2 virus and for commercial success. There would be a reasonable expectation of success given the applied prior art teachings in the art as recited supra. This is analogous to some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the invention as claimed in claims 4-6, 8-9, 12, 14, and 15. See KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007) (see MPEP § 2143, example of rationales, A-G).
22. Relevant Prior Art:
Jurasze et al 2021. Stabilizing the closed SARS-CoV-2 spike trimer. Nat Commun 12, 244.
Dai et al 2020 (CN111592602A, 08/28/2020). Beta coronavirus antigen, preparation method and application thereof.
Zeng et al 2020 (CN111991556A, 11/27/2020). SARS-CoV-2 RBD Conjugated Nanoparticle Vaccine.
Conclusion
23. No claim is allowed.
24. Any inquiry concerning this communication or earlier communications from the examiner should be directed to SAMADHAN J JADHAO whose telephone number is (703)756-1223. The examiner can normally be reached M-F 8:00-5:00.
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/SAMADHAN JAISING JADHAO/ Examiner, Art Unit 1672
/BENNETT M CELSA/Primary Examiner, Art Unit 1600
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