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 with traverse of invention I in the reply filed on 7/9/26 is acknowledged. The traversal is on the ground(s) that inventive Groups 1 and 3 can be examined in view of amendments to at least claim 16 that result in that claim depend from claim 1. In response, claims 1-14 and 16-24 will be examined. However, Group 2 (claim 15 is withdrawn from consideration since that invention doesn’t provide a contribution over the prior art (see below).
The requirement is still deemed proper and is therefore made FINAL.
Claim 15 is withdrawn from further consideration pursuant to 37 CFR 1.142(b), as being drawn to a nonelected invention, there being no allowable generic or linking claim. Applicant timely traversed the restriction (election) requirement in the reply filed on 7/9/26.
Claims 1-14 and 16-24 are examined on the merits.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 1/5/24, 1/31/25, 11/6/25 and 8/25/26 are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 3-5, 13, 16-19, 21, 22 and 24 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.
A broad range or limitation together with a narrow range or limitation that falls within the broad range or limitation (in the same claim) may be considered indefinite if the resulting claim does not clearly set forth the metes and bounds of the patent protection desired. See MPEP § 2173.05(c). In the present instance, claim 3 recites the broad recitation “mitochondrial polymerase”, and the claim also recites “(POLRMT)” which is the narrower statement of the range/limitation. The claim(s) are considered indefinite because there is a question or doubt as to whether the feature introduced by such narrower language is (a) merely exemplary of the remainder of the claim, and therefore not required, or (b) a required feature of the claims.
Claim 17 recites, “a gene segment of VP4 cDNA has a nucleotide sequence of SEQ ID NO: 5 or 6, and a gene segment of VP7 cDNA has a nucleotide sequence of SEQ ID NO: 8, 9…”. However, the recitation of “has a nucleotide sequence of” is indefinite because this limitation includes the sequence of the SEQ ID NO: and fragments thereof. Therefore, it is unclear if fragments are also contemplated as being part of the invention. This rejection can be overcome by amending claim 17 to recite “has the” in place of “has a”.
Claim 18 recites, “a gene segment of VP1 cDNA has a nucleotide sequence of SEQ ID NO: 2…”. Similar limitations are also recited for VP2, VP3, VP6, NSP1, NSP2, NSP3, NSP4 and NSP5. However, the recitation of “has a nucleotide sequence of” is indefinite because this limitation includes the sequence of the SEQ ID NO: and fragments thereof. Therefore, it is unclear if fragments are also contemplated as being part of the invention. This rejection can be overcome by amending claim 18 to recite “has the” in place of “has a”.
Claim 21 recites, “a plasmid consisting of a nucleotide sequence of SEQ ID NO: 20… Similar limitations are also recited for SEQ ID NO:s 21-36. However, the recitation of “has a nucleotide sequence of” is indefinite because this limitation includes the sequence of the SEQ ID NO: and fragments thereof. Therefore, it is unclear if fragments are also contemplated as being part of the invention. This rejection can be overcome by amending claim 21 to recite “consisting of the” in place of “consisting of a”.
Claim 22 recites, “consisting of a nucleotide sequence of SEQ ID NO: 55… Similar limitations are also recited for SEQ ID NO: 56. However, the recitation of “has a nucleotide sequence of” is indefinite because this limitation includes the sequence of the SEQ ID NO: and fragments thereof. Therefore, it is unclear if fragments are also contemplated as being part of the invention. This rejection can be overcome by amending claim 22 to recite “consisting of the” in place of “consisting of a”.
Claim 3 recite a listing of RNA polymerases, however, the grouping is in an improper format according to MPEP 2173.05(h). Therefore, claim 3 is indefinite since the claim structure of “selected from the group consisting of:” is not employed. Since claim 3 does not specify that the grouping of polymerases is closed to those listed, it is unclear if additional polymerases can be read into the claim.
Claims 4 and 5 recite a listing of cell lines, however, the grouping is in an improper format according to MPEP 2173.05(h). Therefore, claims 4 and 5 are indefinite since the claims structure of “selected from the group consisting of:” is not employed. Since claims 4 and 5 do not specify that the grouping of cell lines is closed to those listed, it is unclear if additional cell lines can be read into the claim.
Claim 13 recite a listing of growth medium, however, the grouping is in an improper format according to MPEP 2173.05(h). Therefore, claim 13 is indefinite since the claim structure of “selected from the group consisting of:” is not employed. Since claim 13 does not specify that the grouping of growth medium is closed to those listed, it is unclear if additional growth medium can be read into the claim.
Claim 16 recites a listing of Reovirus gene segments (see lines 7-8), however, the grouping is in an improper format according to MPEP 2173.05(h). Therefore, claim 16 is indefinite since the claim structure of “selected from the group consisting of:” is not employed. Since claim 16 does not specify that the grouping of Reovirus gene segments is closed to those listed, it is unclear if additional Reovirus gene segments can be read into the claim.
Claim 17 recites a listing of serotypes (see line 2), however, the grouping is in an improper format according to MPEP 2173.05(h). Therefore, claim 17 is indefinite since the claim structure of “selected from the group consisting of:” is not employed. Since claim 17 does not specify that the grouping of serotypes is closed to those listed, it is unclear if additional serotypes can be read into the claim.
Claim 17 recites a listing of VP7 cDNA sequences (see line 4), however, the grouping is in an improper format according to MPEP 2173.05(h). Therefore, claim 17 is indefinite since the claim structure of “selected from the group consisting of:” is not employed. Since claim 17 does not specify that the grouping of VP7 cDNA sequences is closed to those listed, it is unclear if additional VP7 cDNA sequences can be read into the claim. Claim 21 also possesses a similar indefiniteness limitation related to the VP7 gene segment.
Claim 24 recites a listing of gene segments (see lines 2-3), however, the grouping is in an improper format according to MPEP 2173.05(h). Therefore, claim 24 is indefinite since the claim structure of “selected from the group consisting of:” is not employed. Since claim 24 does not specify that the grouping of gene segments is closed to those listed, it is unclear if additional gene segments can be read into the claim.
Claim 19 recites the limitation "the plurality of expression vectors" in line 2. There is insufficient antecedent basis for this limitation in the claim.
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)(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.
Claim(s) 1-5, 8-11, 13 and 16-23 are rejected under 35 U.S.C. 102(a)(2) as being anticipated by Wang et al. (US PGPub 2024/0200099).
The claimed invention is drawn to a method for producing a reassortant Reoviridae virus (Rotavirus), the method comprising:
transfecting a BHK21 cell line, into which an T7 RNA polymerase gene is introduced, into an expression vector library comprising a foreign gene;
culturing a first culture by culturing, in a growth medium, the BHK21 cell line undergoing the transfection;
culturing a second culture by adding a MA104 cell line to the first culture that has been cultured; and
culturing a third culture by adding trypsin to the second culture that has been cultured,
wherein the expression vector library comprises a gene segment of cDNA of the Reoviridae virus and a T7 promoter capable of binding to the T7 RNA polymerase. *No specific time limit is required by each of the culturing steps.
The Trypsin added is at a concentration of 1ug/ml and the total DNA weight in the expression vector library is 1ug to 20ug per 1x10^4 to 1x10^6 cells.
The growth media used is DMEM.
The expression vector library comprises a T7 promoter capable of binding to a T7 RNA polymerase and a plurality of expression vectors each comprising a gene segment of rotavirus cDNA operably linked to the T7 promoter, and wherein the plurality of expression vectors comprise a gene segment of cDNA of one of VP1, VP2, VP3, VP4, VP6, VP7, NSP1, NSP2, NSP3, NSP4, and NSP5.
The reassortant rotavirus is a serotype G3 and the cDNA comprises a gene segment of VP4 cDNA has a nucleotide sequence of SEQ ID NO: 5 and a gene segment of VP7 cDNA has a nucleotide sequence of SEQ ID NO: 8. Additionally, the gene segment of a rotavirus cDNA for the VP1, VP2, VP3, VP6, NSP1-5 has a sequence of SEQ ID NO:s 1-4, 7 and 14-18, respectively. The expression cassette further comprises a ribozyme-coding sequence flanked by the gene segment cDNA and the expression cassettes are in a plasmid. At least one plasmid consists of a nucleotide sequence of SEQ ID NO: 20 and comprising the gene segment of VP2. The method also employs a plasmid for expression of D1R consisting of a nucleotide sequence of SEQ ID NO: 55 and a plasmid for expression of D12L consisting of a nucleotide sequence of SEQ ID NO: 56.
Prior Art
Wang et al. teach [0078] “the reverse genetic system contains 11 rotavirus plasmids, each plasmid encoding one of six rotavirus genomic segments (SEQ ID NOs: 1, 3, 5, 7, 9, 11), and 5 helper plasmids that each encode a single helper protein: a fusogenic Fusion-Associated Small Transmembrane (FAST) protein from reovirus (SEQ ID NO: 26); two different RNA capping enzymes from vaccinia virus (SEQ ID NOs: 28, 30); and two additional rotavirus nonstructural proteins NSP2 and NSP5 (SEQ ID NOs: 24-25). The reverse genetics system was modified from Kanai et al. Proc Natl Acad Sci U.S.A. 2017 Feb. 28:114(9):2349-2354 to include additional helper plasmids encoding NSP2 and NSP5 to enhance the formation of viroplasm (see Kawagishi et al. J Virol. 2020 Jan. 6; 94(2): e00963-19). BHK-T7 cells (Buchholz et al., J Virol. 1999 January; 73(1):251-9.) were used for initial virus generation from plasmid transfection, and MA104 cells (Millipore Sigma Cat. #85102918) were co-cultured for virus amplification. After freeze-thaw cycles and trypsin activation, rotavirus was purified by plaque assay on CV-1 cells (CC-170™, American Type Culture Collection (ATCC), Manassas, VA), and further amplified in MA104 cells. Viral seed was confirmed by Sanger sequencing and purified. FIG. 1 is a schematic diagram of the components used to rescue recombinant rotavirus from cells.”
Wang et al. further teach at paragraph [0080] “Sequences of all 16 plasmids used for the generation of wild type SA11 strain were obtained from GenBank Acc. Nos. LC178564-LC178574. pUC19 is the backbone of 11 plasmids, each encoding one rotavirus genome segment insert; pT7/VPISA11 (SEQ ID NO: 1); pT7/VP2SA11 (SEQ ID NO: 3); pT7/VP3SA11 (SEQ ID NO: 5); pT7/VP4SA11 (SEQ ID NO: 7); pT7/VP6SA11 (SEQ ID NO: 9); pT7/VP7SA11 (SEQ ID NO: 11); pT7/NSPISA11 (SEQ ID NO: 13); pT7/NSP2SA11 (SEQ ID NO: 15); pT7/NSP3SA11 (SEQ ID NO: 17); pT7/NSP4SA11 (SEQ ID NO: 19); and pT7/NSP5SA11 (SEQ ID NO: 21). pV1Jns (SEQ ID NO: 23) is the backbone for each of the five helper plasmids, each helper plasmid containing one of the following inserts (inserted using the BglII restriction site): CMV/NSP2 (SEQ ID NO: 24); CMV/NSP5 (SEQ ID NO: 25); CMV/NBVFAST (SEQ ID NO: 26); CMV/D12L (SEQ ID NO: 28), and CMV/DIR (SEQ ID NO: 30). The 2A peptide sequence used was GSGEGRGSLLTCGDVEENPGP (SEQ ID NO: 32). The coding sequences for S1 domain of SARS-COV-2 spike protein (aa Met1-Pro681; SEQ ID NO: 36) and the receptor binding domain (RBD) of SARS-COV-2 spike protein (aa Met1-Cys15 and Arg319-Ser591; SEQ ID NO: 37) were designed based on GenBank: MN908947 (SEQ ID NO: 38), and the SARS-COV-2 spike protein (GenPept: QHD43416; SEQ ID NO: 39). RSV F protein plasmids used were T7/RSV-T4PreF (SEQ ID NO: 42; RSV F protein with DS-Cav1 mutations and T4 foldon at C terminus), T7/RSV-T4scPreF (SEQ ID NO: 45; RSV F protein with linker insertion to prevent furin cleavage, DS-Cav1 mutations, F111 mutations, T4 foldon at C terminus), T7/RSV-A2PreF (SEQ ID NO: 47; RSV F protein with DS-Cav1 mutations), and T7/RSV-A2scPreF (SEQ ID NO: 49; RSV F protein with linker insertion to prevent furin cleavage, DS-Cav1 mutations, F111 mutations); see Zhang et al., Vaccine. 2018 Dec. 18:36(52):8119-8130. All plasmids were synthesized by Genewiz.” And Wang et al. teach the rescue of a specific rotavirus SA11, which is a serotype G3 rotavirus and form the rescue at paragraph [0081] “Recombinant SA1l (rSA11) strains were generated by reverse genetics as described previously with modifications (see Kanai et al., Proc Natl Acad Sci USA. 2017 Feb. 28:114(9):2349-2354). A monolayer of BHK-T7 cells in a 6-well plate (1×10.sup.6 cells/well) was used for transfection. Sixteen plasmids (0.75 μg/plasmid except 0.015 μg pCMV/NSVFAST) were mixed in 150 μl Opti-MEM and added to 150 μl Opti-MEM containing 12.5 μl Lipofectamine2000. Transfection complexes were incubated at room temperature for 20 minutes and then added drop-wise to BHK-T7 cells. 24 hours post transfection, culture medium was exchanged for serum free DMEM. 48 hours post transfection, 1.5×10.sup.5 MA104 cells were added to transfected cells and incubated for 3 days in serum free DMEM supplemented with 1 μg/ml trypsin. To generate recombinant viruses with foreign genes, pT7/NSPISA11, pT7/NSP2SA11, pT7/NSP3SA11, pT7/NSP4SA11, and pT7/NSP5SA11 were replaced with the corresponding plasmid with foreign gene insertion. Where indicated, 450 nucleotides at the 3′ end of the open reading frame were inserted after GFP. rSA11 strains were plaque purified for three rounds using MA104 cells.”
In view of the claim limitations related to specific SEQ ID NO:s, the indefinite limitations of “has a” or “consisting of a” (see 35 112b rejections presented above), the plasmids pT7/VPISA11 (SEQ ID NO: 1); pT7/VP2SA11 (SEQ ID NO: 3); pT7/VP3SA11 (SEQ ID NO: 5); pT7/VP4SA11 (SEQ ID NO: 7); pT7/VP6SA11 (SEQ ID NO: 9); pT7/VP7SA11 (SEQ ID NO: 11); pT7/NSPISA11 (SEQ ID NO: 13); pT7/NSP2SA11 (SEQ ID NO: 15); pT7/NSP3SA11 (SEQ ID NO: 17); pT7/NSP4SA11 (SEQ ID NO: 19); pT7/NSP5SA11 (SEQ ID NO: 21); CMV/NSP2 (SEQ ID NO: 24); CMV/NSP5 (SEQ ID NO: 25); CMV/NBVFAST (SEQ ID NO: 26); CMV/D12L (SEQ ID NO: 28), and CMV/D1R (SEQ ID NO: 30), which possess sequence identity that are fragments with SEQ ID NO:s of claims 17, 18, 21 and 22.
Furthermore, the teachings by Wang et al. at paragraph 81 of “A monolayer of BHK-T7 cells in a 6-well plate (1×10.sup.6 cells/well) was used for transfection. Sixteen plasmids (0.75 μg/plasmid except 0.015 μg pCMV/NSVFAST) were mixed in 150 μl Opti-MEM and added to 150 μl Opti-MEM containing 12.5 μl Lipofectamine2000.” would result in approximately 1.87ug per 1x10^6 cells (0.75 ug x 15 + 0.015 ug = 11.265, divided by 6 wells)
Therefore, Wang et al. anticipate the instant invention.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, 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.
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.
Claim(s) 6, 7, 12, 14 and 24 are rejected under 35 U.S.C. 103 as being unpatentable over Wang et al. as applied to claims 1-5, 8-11, 13 and 16-23 above, and further in view of Komoto et al. (Journal of Virology, 2018, Vol. 92, Issue 13).
The claimed invention also requires when culturing the second culture, the second cell line is added 6 to 12 hours after transfection; and trypsin is added 36 to 48 hours after transfection. The growth medium is supplemented with fetal bovine serum and the media is replaced with growth media without fetal bovine serum prior to adding trypsin (third culture) and this is performed 20 hours after transfection. The molar ratio of the plasmid comprising the gene segment of NSP2 or NSP5 cDNA to the plasmid comprising the gene segment of VP1-VP7 and NSP1, NSP3, NSP4 cDNA is 4:1 to 6:1.
Wang et al. additionally teach that following rotavirus propagation in BHK-T7 and MA104 cells, the resulting cell-virus culture is subjected to freeze-thaw cycling and trypsin treatment and subsequence expansion in MA104 cells (see Figure 1). Therefore, Wang et al. teach the use of trypsin at different times in the propagation of reassortant rotaviruses.
However, Wang et al. do not teach the second cell line is added 6 to 12 hours after transfection; and trypsin is added 36 to 48 hours after transfection. The growth medium is supplemented with fetal bovine serum and the media is replaced with growth media without fetal bovine serum prior to adding trypsin (third culture) and this is performed 20 hours after transfection. The molar ratio of the plasmid comprising the gene segment of NSP2 or NSP5 cDNA to the plasmid comprising the gene segment of VP1-VP7 and NSP1, NSP3, NSP4 cDNA is 4:1 to 6:1.
Komoto et al. teach the generation of reassortant rotaviruses by transfecting BHK cells with plasmids at a ratio 3 to 5 times the amount of NSP2 or NSP5 plasmids to the remaining VP and NSP encoding plasmids (see Table 2 below).
PNG
media_image1.png
440
1790
media_image1.png
Greyscale
The greater amount of NSP2 or NSP5 encoding plasmids teach the claimed molar ratio of plasmids required by claim 24.
It would have been obvious to one of ordinary skill in the art to modify the methods taught by Wang et al. in order to add the MA104 cell line 6 to 12 hours after transfection and to add trypsin 36 to 48 hours after transfection and the growth media is replace 20 hours after transfection and for the molar ratio of the plasmid comprising the gene segment of NSP2 or NSP5 cDNA to the plasmid comprising the gene segment of VP1-VP7 and NSP1, NSP3, NSP4 cDNA is 4:1 to 6:1. One would have been motivated to do so, given the suggestion by Wang et al. that their method of producing a reassortant rotavirus involve the replacement of culture media with serum free DMEM 24 hours after transfection with 11 plasmids (see summary above); and 48 hours after transfection, MA104 cells are added to transfected BHK21 and incubated for 3 days in serum free DMEM supplemented with 1ug/ml trypsin. There would have been a reasonable expectation of success, given the knowledge that reassortant rotaviruses can be generated by transfecting cells with greater amounts (higher molar ratios) of plasmids encoding NSP2 or NSP5, as taught by Komoto et al., and also given the knowledge that MPEP § 2144.05 (II) (A) states, “…Optimization Within Prior Art Conditions or Through Routine Experimentation…“[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to dis-cover the optimum or workable ranges by routine experimentation…”. Therefore, adjusting the time between transfection and growth media replacement and trypsin administration can be optimized by one of ordinary skill in the art. Thus the invention as a whole was clearly prima facie obvious to one of ordinary skill in the art at the time the invention was made.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to BENJAMIN P BLUMEL whose telephone number is (571)272-4960. The examiner can normally be reached M-F 8-5 EST.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Michael Allen can be reached at (571) 270-3497. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/BENJAMIN P BLUMEL/Primary Examiner, Art Unit 1671
Prosecution Insights