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 without traverse of Group I claims 1-6 and 11-16 and a species “a multimer” in the reply filed on 01/16/2026 is acknowledged.
Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement the applicant’s election without traverse is made final.
Status of Claims
3. Claims 1-8 and 11-17 filed as per listing on 01/16/2026 are pending.
4. Claims 7-8 and 17 are withdrawn from consideration due to restriction/election.
5. Claims 1-6 and 11-16 are under examination in this office action.
Priority
6. This application is a U.S. National Phase of International PCT Application No. PCT/CN2021/120518 filed on September 26, 2021, which claims priority to Chinese Patent Application Serial No. 202011351390.9 filed on November 26, 2020.
Information Disclosure Statement
7. Two information disclosure statements (IDSs) submitted on 05/25/2023 and 10/02/2023 was filed in time and the submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Objections to Nucleotide and/or Amino Acid Sequence Disclosures
8. REQUIREMENTS FOR PATENT APPLICATIONS CONTAINING NUCLEOTIDE AND/OR AMINO ACID SEQUENCE DISCLOSURES
Items 1) and 2) provide general guidance related to requirements for sequence disclosures.
37 CFR 1.821(c) requires that patent applications which contain disclosures of nucleotide and/or amino acid sequences that fall within the definitions of 37 CFR 1.821(a) must contain a "Sequence Listing," as a separate part of the disclosure, which presents the nucleotide and/or amino acid sequences and associated information using the symbols and format in accordance with the requirements of 37 CFR 1.821 - 1.825. This "Sequence Listing" part of the disclosure may be submitted:
In accordance with 37 CFR 1.821(c)(1) via the USPTO patent electronic filing system (see Section I.1 of the Legal Framework for Patent Electronic System (https://www.uspto.gov/PatentLegalFramework), hereinafter "Legal Framework") as an ASCII text file, together with an incorporation-by-reference of the material in the ASCII text file in a separate paragraph of the specification as required by 37 CFR 1.823(b)(1) identifying:
the name of the ASCII text file;
ii) the date of creation; and
iii) the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(1) on read-only optical disc(s) as permitted by 37 CFR 1.52(e)(1)(ii), labeled according to 37 CFR 1.52(e)(5), with an incorporation-by-reference of the material in the ASCII text file according to 37 CFR 1.52(e)(8) and 37 CFR 1.823(b)(1) in a separate paragraph of the specification identifying:
the name of the ASCII text file;
the date of creation; and
the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(2) via the USPTO patent electronic filing system as a PDF file (not recommended); or
In accordance with 37 CFR 1.821(c)(3) on physical sheets of paper (not recommended).
When a “Sequence Listing” has been submitted as a PDF file as in 1(c) above (37 CFR 1.821(c)(2)) or on physical sheets of paper as in 1(d) above (37 CFR 1.821(c)(3)), 37 CFR 1.821(e)(1) requires a computer readable form (CRF) of the “Sequence Listing” in accordance with the requirements of 37 CFR 1.824.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed via the USPTO patent electronic filing system as a PDF, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the PDF copy and the CRF copy (the ASCII text file copy) are identical.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed on paper or read-only optical disc, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the paper or read-only optical disc copy and the CRF are identical.
Specific deficiency – Nucleotide and/or amino acid sequences appearing in the specification are not identified by sequence identifiers in accordance with 37 CFR 1.821(d).
Required response – Applicant must provide:
A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3) and 1.125 inserting the required sequence identifiers, consisting of:
A copy of the previously submitted specification, with deletions shown with strikethrough or brackets and insertions shown with underlining (marked-up version);
A copy of the amended specification without markings (clean version); and
A statement that the substitute specification contains no new matter.
The instant claim 11 and instant specification recites NCBI Accession No. for the protein sequences (e.g. AEI61557.1) instead of the SEQ ID NO in compliance with the sequence claims and submission guidelines.
Objections to Specification
9. The use of the term “NCBI”, recited in the instant specification (05/26/2023) on pages 3-6, and page 8 which is a trade name, or a mark used in commerce, has been noted in this application. The term should be accompanied by the generic terminology; furthermore the term should be capitalized wherever it appears or, where appropriate, include a proper symbol indicating use in commerce such as ™, SM , or ® following the term.
Although the use of trade names and marks used in commerce (i.e., trademarks, service marks, certification marks, and collective marks) are permissible in patent applications, the proprietary nature of the marks should be respected and every effort made to prevent their use in any manner which might adversely affect their validity as commercial marks. Applicant is required to properly annotate all trade names and/or marks present in the instant specification, if any additional trade names and/or marks are discovered. Applicant' s cooperation is requested in correcting any errors of which applicant may become aware in the specification.
10. The specification is objected to by providing more than six foreign reference citations (submitted as Foreign References on 05/25/2023) that are not of record and/or English translated (in part or whole) as relevant to the disclosed teaching. For example, Applicant referenced the Chinese language Patent Application No. 202011351390.9 filed on November 26, 2020, as recited in “Cross Reference to Related Applications” in the 05/26/2023 preliminary amendment (instant specification page 1), patent CN 101293918 B (instant specification page 13), patent CN 101148661 B (on specification page 14, 17), patents CN 101293918 B, CN 1976718 A (on specification page 16), and additionally, elsewhere may be recited in the specification. Applicant may provide copies of English translated portions or entire prior art documents or at least machine English translations of the Chinese prior arts (e.g. google machine English translation) for clarity of record.
11. The specification recites the sequence identified by NCBI sequence accession number AEI61557.1, ABU55797.1, AEI61589.1, AIF71344.1, APQ44868.1, AEI61581.1, AIF71350.1, CAD1814034.1, etc. in NCBI database (See, page numbers 3, 4-6, and 8, and additionally may be other pages of specification). NCBI® accession numbers are essential material and are required by the instant claims by reference to a publication. Incorporation of essential material by reference to a publication is improper under 37 CFR § 1.57(d). Applicant is required to amend the disclosure to include the material incorporated by reference. 37 CFR § 1.57. The amendment must be accompanied by an affidavit or declaration executed by the applicant, or a practitioner representing the applicant, stating that the amendatory material consists of the same material incorporated by reference in the referencing application. See In re Hawkins, 486 F.2d 569, 179 USPQ 157 (CCPA 1973).
Furthermore, if the recited NCBI accession AEI61557.1, ABU55797.1, AEI61589.1, AIF71344.1, APQ44868.1, AEI61581.1, AIF71350.1, CAD1814034.1, etc. in NCBI database in instant claim 11, was not set forth in the specification as filed, and was not publicly available at the time the application was filed, the amendment will be treated as an attempt to introduce new matter (similar to attempts to incorporate essential material by reference to unpublished material). In addition, note that that an amendment introducing an additional sequence corresponding to the intended sequence will probably require a replacement Sequence Listing, CRF and a statement indicating that the paper and CRF sequence submissions are the same.
Claim Rejections - 35 USC § 112
12. 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.
The instant claim 11 is 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 claim 11 contains the trademark/trade name NCBI. Here, NCBI corresponds to an entity and not content and a such functions in the same manner as a “trade name” or “trade mark”. Where a trademark or trade name is used in a claim as a limitation to identify or describe a particular material or product, the claim does not comply with the requirements of 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph. See Ex parte Simpson, 218 USPQ 1020 (Bd. App. 1982). The claim scope is uncertain since the trademark or trade name cannot be used properly to identify any particular material or product. A trademark or trade name is used to identify a source of goods, and not the goods themselves. Thus, a trademark or trade name does not identify or describe the goods associated with the trademark or trade name. In the present case, the trademark/trade name is used to identify/describe “a protein or amino acid sequence identifier, SEQ ID NO”.
In addition, in the claim 11 the recited NCBI® accession No. AEI61557.1, ABU55797.1, AEI61589.1, AIF71344.1, APQ44868.1, AE161581.1, AIF71350.1, and CAD1814034.1 for the requisite sequences fail to clearly identify the precise HPV52 L1 protein sequences claimed because the accession numbers are extraneous to the instant disclosure.
Accordingly, the claimed identification/description of the HPV52L1 sequences in the instant claim 11 is indefinite.
Claim Interpretation
13. 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 interpreted to be directed to a modified HPV52 L1 protein comprising a modification selected from the group consisting of the following or a combination thereof, when compared with a wild-type HPV52 L1 protein:
mutating the amino acid at position 447 from aspartate to glutamate;
deleting 1 to 20 successive or nonsuccessive amino acids at the N-terminus;
deleting 1 to 25 successive or nonsuccessive amino acids at the C-terminus;
substituting one or more amino acids at positions 1 to 20 at the N-terminus; and
substituting one or more amino acids at positions 1 to 25 at the C-terminus.
The elected claim 1 and independent claims 2-6 and 11-16 are interpreted to be directed to modifications (deletion, insertion, substitution) in the amino acid sequences at the N-terminal and or C-terminal end of L1 protein of HPV52 to improve expression, solubility and purification yield of the expressed truncated L1 protein of HPV52 to obtain multimeric (pentamers) protein or VLPs for diagnostics antigen and immunogen applications. As deemed necessary, the modified HPV52L1 protein encoding polynucleotide is codon optimized to express for higher yield in insect cells sf9 or yeast cells or prokaryotic E. coli, and expression vector plasmid, baculovirus vector is used to express the protein in a compatible cell culture system.
Claim Rejections - 35 USC § 102
14. 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 the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
15. Claims 1, 2, 12, and 13 are rejected under 35 U.S.C. 102(a)(1)/(a)(2) as being anticipated by Li et al 2016 (US9499591B2, 11/22/2016).
Claim 1: Li et al 2016 (US9499591B2) anticipated instant claim 1 by disclosing a truncated (reads on modified) HPV52 L1 protein comprising a modification wherein the truncated HPV52 L1 protein is different from wild type HPV52 L1 protein by a deletion of amino acid positions 2-35, 2-40, or 2-42 at an N-terminal of the wild type HPV52 L1 protein (See, abstract, claim 1). Li et al 2016 anticipated a Markush group limitation: deleting 1 to 20 successive or non-successive amino acids at the N-terminus.
Claim 2. The modified HPV52 L1 protein according to claim 1, which is as shown in the sequence selected from SEQ ID No. 2 to SEQ ID No. 29.
Li et al 2016 (US9499591B2) is directed to a truncated L1 protein of Human Papillomavirus Type 52 and has disclosed SEQ ID NO: 12 that has 99.9% amino acid identity with instant SEQ ID NO: 29 as recited below. A single amino acid difference in identity is due to a single conservative amino acid substitution in the reference database D434E of instant claimed SEQ ID NO: 29.
Query Match 99.9%; Score 2514; Length 490;
Best Local Similarity 99.8%;
Matches 464; Conservative 1; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVSGLQY 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVSGLQY 60
Qy 61 RVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLLNKFDDTE 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 RVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLLNKFDDTE 120
Qy 121 TSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNSGNPGDCPPLQL 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 TSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNSGNPGDCPPLQL 180
Qy 181 INSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQMASEPYGDSLFFFL 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 INSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQMASEPYGDSLFFFL 240
Qy 241 RREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSAFFPTPSGSMVTSESQLFN 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 RREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSAFFPTPSGSMVTSESQLFN 300
Qy 301 KPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEE 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 KPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEE 360
Qy 361 FDLQFIFQLCKITLTADVMTYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQ 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 FDLQFIFQLCKITLTADVMTYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQ 420
Qy 421 KNTPPKGKEDPLKEYMFWEVDLKEKFSADLDQFPLGRKFLLQAGL 465
|||||||||||||:|||||||||||||||||||||||||||||||
Db 421 KNTPPKGKEDPLKDYMFWEVDLKEKFSADLDQFPLGRKFLLQAGL 465
Claim 12: Li et al 2016 (US9499591B2) anticipated the instant claim 12 by disclosing the added limitation, wherein the wild-type HPV52 L1 protein is as shown in SEQ ID No. 1. As recited below Li et al 2016 disclosed below the prior art SEQ ID NO: 1 that has 100% amino acid identity with instant SEQ ID NO: 1:
Query Match 100.0%; Score 2712; Length 503;
Best Local Similarity 100.0%;
Matches 503; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNG 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNG 60
Qy 61 KKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGI 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 KKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGI 120
Qy 121 SGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNN 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 SGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNN 180
Qy 181 NSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQM 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 NSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQM 240
Qy 241 ASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSAFFPTP 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 ASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSAFFPTP 300
Qy 301 SGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLCAEVKKESTYK 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 SGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLCAEVKKESTYK 360
Qy 361 NENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILEDWQFGLTPPPSASLED 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 NENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILEDWQFGLTPPPSASLED 420
Qy 421 TYRFVTSTAITCQKNTPPKGKEDPLKDYMFWEVDLKEKFSADLDQFPLGRKFLLQAGLQA 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 TYRFVTSTAITCQKNTPPKGKEDPLKDYMFWEVDLKEKFSADLDQFPLGRKFLLQAGLQA 480
Qy 481 RPKLKRPASSAPRTSTKKKKVKR 503
|||||||||||||||||||||||
Db 481 RPKLKRPASSAPRTSTKKKKVKR 503
Claim 13. Li et al 2016 (US9499591B2) anticipated instant claim 13 by disclosing the modified HPV52 L1 protein according to claim 1, deleting 2, 4, 5, 8, 10, 13, 14, 15, 18, or 20 successive or nonsuccessive amino acids at the N-terminus (See, abstract, claim 1).
Claim Rejections - 35 USC § 103
16. 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.
17. Claims 1, 2, 3-6, 11-12, and 13-14 are rejected under 35 U.S.C. 103 as being unpatentable over Li et al 2016 (US9499591B2, 11/22/2016) as applied to claim 1, 2, 12, and 13 above, and further in view of Godi et al 2018 (J Gen Virol. 2019 Feb;100(2):237-245), Sun et al 2016 (Appl Microbiol Biotechnol (2016) 100:1231–1240), Wei et al 2018 (Emerg Microbes Infect. 2018 Sep 26;7(1):160), Ma et al 2008 (CN101245099A, 08/20/2008), Bhat et al 2018 (Glob J Biotechnol Biomater Sci. 2017; 4(1): 001-007), and The Protein Man’s Blog 2018 (PDF printout, G-Biosciences, Geno Technology Inc, Online publication, info.gbiosciences.com/blog/tips-for-preventing-protein-aggregation-loss-of-protein-solubility).
Claims 1, 2, 3-6, 12, and 13-14: Li et al 2016 (US9499591B2) anticipated by instant claims 1, 2, 12, and 13 as recited supra and the teachings are incorporated here in its entirety.
Li et al 2016 (US9499591B2) is in the art and teaches a truncated L1 protein of Human Papillomavirus Type 52, its coding sequence and preparation method, and a virus-like particle (VLP) comprising the protein, wherein the protein and the VLPs are useful for preventing HPV (particularly HPV52) infection, and a disease caused by HPV (particularly HPV52) infection, such as cervical cancer. The invention also relates to the use of the protein and the VLPs in the preparation of a pharmaceutical composition or a vaccine for preventing HPV (particularly HPV52) infection, and a disease caused by HPV (particularly HPV52) infection, such as cervical cancer. A truncated HPV52 L1 protein, wherein the truncated HPV52 L1 protein is different from wild type HPV52 L1 protein by a deletion of amino acid positions 2-35, 2-40, or 2-42 at an N-terminal of the wild type HPV52 L1 protein, wherein said truncated HPV52 L1 protein (See, abstract, claim 1). Li et al 2016 disclosed a truncated L1 protein of Human Papillomavirus Type 52 SEQ ID NO: 12 that has 99.9% amino acid identity with instant SEQ ID NO: 29 as recited below. A single amino acid difference in identity is due to a single conservative amino acid substitution in the reference database D434E of instant claimed SEQ ID NO: 29 (See, above 35 USC 102 anticipation rejection for claim 2 truncated HPV52L1 amino acid sequence, and claim 12 wild type HPV52L1 amino acid sequence).
Li et al 2016 does not teach an alternative limitation of instant claim 1, D447E substitution, HPVL52L1 polynucleotide codon optimization for insect cell expression of the protein.
Godi et al 2018 is in the art of HPV52 L1 VLP and teaches an HPV 52 lineage D L1 protein containing D447E, VLPs (VLPs reads on multimer or pentamer), and codon-optimized gene, vectors, cells, and L1 and L2 pseudoviruses thereof. Homology models of the HPV52 L1 pentamer were generated which permitted mapping these residues to a small cluster on the outer rim of the surface exposed pentameric L1 protein. (See, Godi et al 2018, J Gen Virol. 2019 Feb;100(2):237-245, full article, pages 240 and 243, and figures 1-2, abstract, as recited in PCT written opinion on file (05/25/2023).
Claims 3-6: Li et al 2016 (US9499591B2) as recited supra teaches the modified HPV52 L1 protein according to claim 1. Li et al 2016 also teaches (instant claim 2 above) a coding sequence (reads on polynucleotide) of the truncated HPV52 L1 protein, a virus-like particle (VLP) comprising the protein, and a method of preparing the protein and the VLP. Li et al 2016 further disclosed SEQ ID NO: 12 that has 99.9% amino acid identity with instant SEQ ID NO: 29 as recited supra (with a single a single conservative amino acid substitution as compared to the prior art reference sequence “Aspartate” D434E “Glutamate” of instant claimed SEQ ID NO: 29. Li et al 2016 further teaches (the instant claim 12) the wild-type HPV52 L1 protein amino acid sequence (the prior art SEQ ID NO: 1) that has 100% amino acid identity with instant SEQ ID NO: 1 that is a wild type HPV52 L1 protein. Li et al 2016 teaches eukaryotic expression systems insect baculovirus vector system for HPV L1 protein expression, HPV52 virus-like particle, comprising or consisting of or formed from the truncated protein, and VLP production.
Li et al 2016 teaches a host cell comprising the polynucleotide or vector. The host cell includes but is not limited to prokaryotic cells such as E. coli cells, and eukaryotic cells such as yeast cells, insect cells. The host cell according to the invention may also be a cell line, such as 293T cell. Li et al 2016 teaches plasmid p52L1h, the pAAV vector carrying the nucleotide sequence encoding HPV52 L1 protein. Li et al 2016 teaches the expression plasmid for expressing HPV structural proteins was codon-optimized to express HPV L1 gene efficiently in mammalian cells, thereby facilitating highly efficient assembly of pseudo-virion (reads on VLPs).
Li et al 2016 (US9499591B2) disclosed SEQ ID NO: 14 that has 64.7% nucleotide sequence identity (78% best local similarity) with instant SEQ ID NO: 31 of instant claim 4 (instant claim 4).
Li et al 2016 teaches the polynucleotide sequence to express the claimed HPV52L1, insect cells for expression, and baculovirus vector. However, does not teach codon optimized polynucleotide whole-gene HPV52L1 optimized using insect cell codons.
Sun et al 2016 is in the HPVL1 VLP art and teaches a 21 amino acid NLS lacking C-terminally truncated HPV6 L1 polynucleotide sequence gene that is optimized for insect cell codons expression (codon optimized for insect cells) and production of the VLPs (See, abstract, page 1238 col 1 last para, page 1232 col 2 para 1, Fig. 6 a, b).
Claims 11-12:
Claim 11: The modified HPV52 L1 protein according to claim 1, wherein the wild-type HPV52 L1 protein is as shown in the sequence selected from the group consisting of NCBI Accession No. AEI61557.1, ABU55797.1, AEI61589.1, AIF71344.1, APQ44868.1, AE161581.1, AIF71350.1, and CAD1814034.1.
Claim 12: The modified HPV52 L1 protein according to claim 1, wherein the wild-type HPV52 L1 protein is as shown in SEQ ID No. 1.
Examiner’s note regarding claim 11: The recited NCBI® designations for the requisite sequences fail to clearly identify the precise HPV52 L1 protein sequences claimed because the accession numbers are extraneous to the instant disclosure. In the interest of compact prosecution, claim 11 is interpreted as HPV52 L1 wild-type protein sequences.
Claim 12: Li et al 2016 (US9499591B2) teaches the instant claim 12 by disclosing the added limitation, wherein the wild-type HPV52 L1 protein is as shown in SEQ ID No. 1. As recited below Li et al 2016 disclosed below the prior art SEQ ID NO: 1 that has 100% amino acid identity with instant SEQ ID NO: 1:
Query Match 100.0%; Score 2712; Length 503;
Best Local Similarity 100.0%;
Matches 503; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNG 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNG 60
Qy 61 KKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGI 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 KKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGI 120
Qy 121 SGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNN 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 SGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNN 180
Qy 181 NSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQM 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 NSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQM 240
Qy 241 ASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSAFFPTP 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 ASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSAFFPTP 300
Qy 301 SGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLCAEVKKESTYK 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 SGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLCAEVKKESTYK 360
Qy 361 NENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILEDWQFGLTPPPSASLED 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 NENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILEDWQFGLTPPPSASLED 420
Qy 421 TYRFVTSTAITCQKNTPPKGKEDPLKDYMFWEVDLKEKFSADLDQFPLGRKFLLQAGLQA 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 TYRFVTSTAITCQKNTPPKGKEDPLKDYMFWEVDLKEKFSADLDQFPLGRKFLLQAGLQA 480
Qy 481 RPKLKRPASSAPRTSTKKKKVKR 503
|||||||||||||||||||||||
Db 481 RPKLKRPASSAPRTSTKKKKVKR 503
Claims 13-14:
Claim 13: The modified HPV52 L1 protein according to claim 1, deleting 2, 4, 5, 8, 10, 13, 14, 15, 18, or 20 successive or nonsuccessive amino acids at the N-terminus.
Wei et al 2018 is in the art and teaches added limitation of instant claim 13 by disclosing N-terminal truncations on L1 proteins of human papillomaviruses promote their soluble expression in Escherichia coli and self-assembly in vitro. We et al 2018 teaches up to 15 amino acid truncations at the N-terminal of HPV 52 L1 protein achieved increased expression and solubility for truncated protein constructs N5C, N10C, N15C and N19C. The nucleotide sequences, vectors, host cells, virus-like particles, vaccines and the like thereof, wherein N-terminal truncations can promote the expression of the L1 proteins are taught. In our previous work, we found that truncation of several N-terminal residues could improve proteins the solubility of HPV L1 (See, abstract, result, page 9, and figure 1 and legends). It would have been obvious to try for one of the ordinary skills to truncate the HPV52L1 N-terminal protein based on the teachings of Wei et al 2018 and attempt to obtain increased expression and solubility in eukaryotic expression system (e.g. insect cells, mammalian cells). It would have been "Obvious to try" to the ordinary skills by choosing from a finite number of identified, predictable solutions, with a reasonable expectation of success by deleting 2, 4, 5, 8, 10, 13, 14, 15, 18, or 20 successive or non-successive amino acids at the N-terminus. (See, MPEP § 2143, example of rationales, A-G).
Ma et al 2008 (CN101245099A, 08/20/2008) discloses a recombinant HPV L1 capsid protein, being capable of being dissolved in water and expressed to obtain an Ll pentamer having the same immunogenicity and antigenicity as a wild type protein. Compared with the wild type protein, the N-terminal is substituted and the C-terminal is truncated. The amino acid sequence of the HPV52 L1 protein is shown in SEQ ID NO: 13 (see, claims 1-12, description).
Claim 14: The modified HPV52 L1 protein according to claim 1, deleting 13 amino acids at the N-terminus and substituting with any one selected from the group consisting of serine, serine-glutamate, serine-glutamate-arginine, and proline-serine-glutamate-alanine-threonine.
Bhat et al 2018 is in the protein expression and purification art. Bhat et al 2018 teaches the protein solubility is achieved by the site-directed mutagenesis that generates the hydrophobic to hydrophilic mutations, a hydrophobic composition of protein adversely affects solubility, delete or mutate the hydrophobic residues which increase the protein expression and solubility. Alpha helices are stabilized by predominantly hydrogen bonds, while hydrogen bonding and hydrophobic interactions are the stabilizing forces for beta sheets. Exposed hydrophobic residues in a protein molecule in the cell due to misfolding or mutation tend to stick together and lead to the formation of insoluble aggregates called inclusion bodies in recombinant proteins (See, abstract, entire article).
Protein Man’s Blog 2018 is in the protein art and teaches adding a mixture of arginine and glutamate to your buffer increases protein solubility by directly binding to charged and hydrophobic regions (See, PDF printout).
Therefore, to arrive at the added limitations of claims 13-14, it would have been "Obvious to try" to the ordinary skills by combining the prior art teachings Wei et al 2018, Bhat et al 2018, and Protein Man’s Blog 2018 by choosing from a finite number of identified, predictable solutions, with a reasonable expectation of success by deleting 13 amino acids at the N-terminus and substituting N-terminal hydrophobic (non-polar) amino acids with hydrophilic (polar) amino acids and balance the hydrophobic and hydrophilic properties of the N-terminal domain of HPV52L1 protein with the alternative claimed limitations: any one selected from the group consisting of serine, serine-glutamate, serine-glutamate-arginine, and proline-serine-glutamate-alanine-threonine. (See, MPEP § 2143, example of rationales, A-G).
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 Li et al 2016 as applied to claim 1 and relevant dependent claims with additional teachings of Godi et al 2018 on D447E mutation, and Sun et al 2016 on codon optimization and baculovirus expression for the HPV52 L1 polynucleotide sequence to produce the HPV52 L1 multimer protein or the HPV52 L1 VLPs taught by the prior arts Li et al 2016, HPV52L1 protein N-terminal 15 amino acid deletions by both Wei et al 2018, and Ma et al 2008, hydrophobic amino acid substitutions or deletion approach taught by Bhat et al 2018, and the insight provided by Protein Man’s Blog 2018 on hydrophobic protein property neutralization by addition of hydrophilic amino acid (e.g. mixture of arginine and glutamate) addition to increase solubility of protein to arrive at the inventions of claims 1-6, 11-12, and 13-14. One of the ordinary skills in the art would have been motivated to develop a codon optimized baculovirus expression vector for expression of the claimed HPV52L1 in the insect cells (e.g. Sf9) for producing soluble protein for purification, HPV52 L1 VLPs for production of immunogens, vaccines and diagnostic antigens 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 1-6, 11-12, and 13-14. 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).
18. Claims 15-16 are rejected under 35 U.S.C. 103 as being unpatentable over combined teachings of Li et al 2016 (US9499591B2, 11/22/2016), Godi et al 2018 (J Gen Virol. 2019 Feb;100(2):237-245), Sun et al 2016 (Appl Microbiol Biotechnol (2016) 100:1231–1240), Wei et al 2018 (Emerg Microbes Infect. 2018 Sep 26;7(1):160), Ma et al 2008 (CN101245099A, 08/20/2008), Bhat et al 2018 (Glob J Biotechnol Biomater Sci. 2017; 4(1): 001-007), and The Protein Man’s Blog 2018 (PDF printout, G-Biosciences, Geno Technology Inc, Online publication, info.gbiosciences.com/blog/tips-for-preventing-protein-aggregation-loss-of-protein-solubility) as applied to claims 1, 2, 3-6, 11-12, and 13-14 above, and further in view of Debruus et al 2007 (CN1976718A, 06/06/2007), Yang et al 2006 (Genomics Proteomics Bioinformatics. 2006 Feb;4(1): p. 34-41), Wu et al 2015 (CN104710515A, published 06/17/2015, English translated PDF printout), Kong et al 2015 (CN104418942A, 03/18/2015, English translated PDF printout), Zhou et al 1991 (Virology. 1991 Dec;185(2):625-32), and Chen et al 2001 (J Mol Biol. 2001 Mar 16;307(1):173-82).
Claims 15-16: The combined prior art teachings as applied to claims 1, 2, 3-6, 11-12, and 13-14 above teaches claim 1 as recited supra, however does not teach added limitations of instant claims 15-16.
Claim 15. The modified HPV52 L1 protein according to claim 1, deleting 19 or 25 successive or nonsuccessive amino acids at the C-terminus.
Claim 16. The modified HPV52 L1 protein according to claim 1, substituting one or more basic amino acids at positions 1 to 23 at the C-terminus with anyone selected from the group consisting of polar uncharged amino acid, non-polar amino acid, and acidic amino acid;
wherein the basic amino acid is selected from arginine and/or lysine;
wherein the polar uncharged amino acid is selected from the group consisting of glycine,
serine, and threonine;
wherein the non-polar amino acid is selected from alanine and/or valine; and
wherein the acidic amino acid is selected from aspartate and/or glutamate.
Debruus et al 2007 (CN1976718A) disclosed a C-terminally truncated Ll protein. Truncation eliminates less than 40 amino acids, and suitable truncation of HPV52 can be performed so as to suitably remove the equivalent C terminal portion of the described L1 protein (see description, pages 6-7).
Yang et al 2006 used bioinformatic analysis to predict the nuclear localization signals (NLS) of 107 types of HPV L1 proteins including HPV52 L1 protein (See, abstract, Table 1 page 37 col 1 HPV 52, entire prior art) and thus Yang et al 2006 teaches that NLS are largely conserved in different HPV types in the 31 amino acids at the C-terminal.
Wu et al 2015 (CN104710515A) is in the art and discloses an approach or method of a non-basic amino acid substitution with another non-basic amino acid (non-polar amino acid) in human papilloma virus L1 protein mutants and a preparation method thereof, and a pharmaceutical composition containing at least one of the mutants, especially a vaccine containing at least one of the mutants (See, abstract). A human papillomavirus L1 protein refers to HPV types 6, 11, 16, 18, 31, 33, 34, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 67, 68, 70, 72, 73 or 82, 85, 97, HPV types and/or a combination of several. A mutant of human papilloma virus L1 protein, (reads on HPV11 L1 protein) which is characterized in that the leucine L* in the C-terminal α-helix 5 FPLGRKFL*LQAG of the human papillomavirus L1 protein is mutated (mutated reads on substituted) into alanine A (non-polar amino acid) or serine S (polar amino acid), preferably mutated to alanine A (non-polar amino acid). The human papillomavirus (HPV) L1 protein refers to HPV11 type L1 (See, CN104710515A, English translated PDF print out, page 4, Contents of Invention, lines 1-7, claims 1-2).
Kong et al 2015 (CN104418942A) is in the art and teaches C-terminal 9 amino acids or 21 amino acids truncated L1 proteins and nucleotide sequences encoding these truncated L1 proteins of HPV, and VLPs for HPV11-type, and including HPV types HPV52 (See, Description section on Determination of Pseudovirus Virus Titer), production of the truncated proteins using E coli (prokaryotic vector), eukaryotic cells and eukaryotic vector, baculovirus vector and insect cells, Saccharomyces cerevisiae yeast and yeast vector (See, Kong et al 2015, CN104418942A, English translated PDF print out, abstract, claim 1, page 4).
Further, the following prior art references provide additional prior art recognition of the structural and functional equivalency of the L1 protein of HPV52 and additional prior arts on the HPV11 and HPV16 strains that would motivation one of ordinary skill in the art to modify the primary prior art reference applied to claim 1 above for instant claims 15-16 C-terminal basic modifications specifically applied to Zhou et al for HPV16 L1, and Chen et al 2001 for the suggested HPV16 L1 and HPV11L1 embodiment.
Zhou et al 1991 is in the HPV 16 L1 protein art and teaches identification of the nuclear localization signal of human papillomavirus using HPV16 L1 protein as a model. Human papillomavirus type 16 (HPV16) L1 and L2 capsid proteins can be detected only in the nucleus of infected cells. For other proteins that accumulate or destined to be transported to the nucleus are called nuclear proteins, and those nuclear proteins have specific sequences of basic amino acids(aa) termed nuclear localization signals (NLS) that direct the protein from the cytoplasm to the nucleus. A series of deletion and substitution mutations in the HPV16 L1 protein, produced by recombinant vaccinia virus (rVV), to identify NLS within HPV16 L1 and showed that HPV16 L1 C-terminal contains two NLS sequences, each containing basic amino acid clusters. One NLS consisted of 6 basic amino acids (KRKKRK from aa 525 to 530) at the carboxy terminal end of L1. The other NLS contained 2 basic amino acid clusters (KRK from aa 510 to 512 and KR at aa 525, 526) separated by 12 amino acids. Mutations in either one of the NLS basic amino acid clusters did not alter nuclear localization of L1 when the other NLS basic amino acid cluster remained intact, but mutations to NLS basic amino acid clusters prevented nuclear localization of L1 protein. The L1 NLS and nuclear localization could be overridden by introduction of a membrane binding sequence MVVLLAALLVV (non-basic amino acid) at the amino terminal end of the protein. A databases search showed that all sequenced papillomaviruses are predicted to have L1 and L2 capsid proteins with sequences of basic amino acids homologous with one or both NLS of HPV16 L1. (See, Zhou et al 1991, abstract, page 626 col 2 last para continued to page 627, Results col 2 on page 627 last 2 para, discussion, entire article). This Zhou et al 1991 teaches insertional mutation of non-polar amino acids, and deletion of a patch of amino acids at the C-terminal of HPV16 L1 protein that results in loss of nuclear localization of the L1 protein.
Zhou et al 1991 teaches substitution of a cluster of basic amino acids (more than one amino acids) in the NLS at C- terminal of HPV16 L1 with nonpolar amino acids, however, does not teach substitution of a single basic amino acid at C-terminus modified HPV11 L1 protein of 31 amino acids at C-terminus that are substituted with amino acids selected from the group consisting of: polar uncharged amino acids, non-polar amino acids, and acidic amino acids.
Zhou et al 1991 teaches the inventive concept of disruption of NLS basic amino acid cluster by deletion or substation of cluster of a basic amino acid to enable to direct the HPV16 L1 protein to the cytoplasm. Zhou et al 1991 in table 3 (page 631, col 1, lower half of table) shows conserved basic amino acid sequences in the NLS at C-terminal for HPV16 L1, HPV11 L1 and for L1 protein of other HPV types. Therefore, one of the ordinary skills would have been apprised of the importance of the basic amino acid clusters or basic amino acids at the C-terminal for 31 amino acids that determine the NLS and fate of the L1 protein to transport from cytoplasm to the cell nucleus to localize in the nucleus among different HPV serotypes irrespective of the HPV serotype. Therefore, following this prior art would motivate one to substitute, for virally related protein strains, the C-terminal basic amino acid sequences for disruption of L1 protein NLS in HPV11 L1 protein to produce substantially large quantity of soluble L1 protein or VLPs by retaining the produced protein with disrupted NLS within the cytoplasm.
Chen et al 2001 is in the art and teaches that the L1 major capsid proteins of human papillomavirus HPV 11 and HPV 16 were purified and analyzed for structural integrity and in vitro self-assembly. Proteins were expressed in Escherichia coli as glutathione-S-transferase-L1 (GST-L1) fusions and purified to near homogeneity as pentamers (equivalent to viral capsomeres), after thrombin cleavage from the GST moiety and removal of tightly associated GroEL protein. Sequences at the amino and carboxy termini contributing to formation of L1 pentamers and to in vitro capsid assembly were identified by deletion analysis. For both HPV11 and HPV16 L1, up to at least ten residues could be deleted from the amino terminus (Delta N10) and 30 residues from the carboxy terminus (Delta C30) without affecting pentamer formation. The HPV16 pentamers assembled into relatively regular, 72-pentamer shells ("virus-like particles" or VLPs) at low pH, with the exception of HPV16 L1 Delta N10, which assembled into a 12-pentamer, T=1 capsid (small VLP) under all conditions tested. The production of large quantities of assembly-competent L1, using the expression and purification protocol described here, has been useful for crystallographic analysis, and will be valuable for studies of virus-receptor interactions and potentially for vaccine design (See, abstract). For example, the similarity of HPV16 and HPV11 L1 sequences ceases abruptly 30 residues from the carboxy terminus, just C-terminal to helix h5 (See, page 180, col 1 para 4). The C-terminus 26 amino acid residues deleted at encompass deletions of basic amino acids, lysine and arginine residues (upper case) conserved in papillomavirus L1 proteins, according to Table 3 of Zhou et al 1991 page 631 col 1.
To the extent that the Zhou et al 1991 and Chen et al 2001 reference method discussed above does not expressly teach a “single” mutant L1 basic amino C-terminal substitution, the Wu et al 2015 reference cited above teach that changing “one or more” basic amino acid represent an obvious design choice.
Therefore, based on the prior art teachings recited supra, it would have been obvious to one of the ordinary skills in the art to disrupt the HPV52 L1 NLS by deleting 19 or 25 successive or nonsuccessive amino acids at the C-terminus (instant claim 14 limitation) or by substituting one or more basic amino acids at positions 1 to 23 at the C-terminus with anyone selected from the group consisting of polar uncharged amino acid, non-polar amino acid, and acidic amino acid (instant claim 16 limitation) to prevent nuclear localization of the expressed protein and obtaining substantially higher amount of soluble HPV52 L1 protein as compared to the wild type HPV52 L1 protein.
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 Li et al 2016 as applied to claim 1 and with additional teachings of the prior arts as applied to claims 15-16 above to arrive at the inventions of claims 15-16. One of the ordinary skills in the art would have been motivated to develop a codon optimized baculovirus expression vector for expression of the claimed HPV52L1 in the insect cells (e.g. Sf9) for producing soluble protein for purification, HPV52 L1 VLPs for production of immunogens, vaccines and diagnostic antigens 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 15-16. 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).
18. Relevant Prior Arts:
Merck Sharp and Dohme BV. (CN 1934131 A, 21 March 2007). Optimal Expression of HPV52 L1 in Yeast.
Arifah et al 2025. Truncation on N-Terminal Hydrophobic Domain of L1 Major Capsid Protein of Human Papillomavirus Type 52 Enhances Its Expression in Hansenula polymorpha. HAYATI Journal of Biosciences 32(4):1062-1072 (Year: 2025)
Conclusion
19. No claim is allowed.
20. 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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