DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Applicant’s amendment and response filed on 2/5/2026 have been received and entered into the case. Claims 1-16, 31-32 and 40 have been canceled, Claim 41 has been added. Claims 17-30, 33-39 and 41 are pending, Claims 19-20, 23, 25-27, 29, 33-34, 36-39 and 41 have been withdrawn, and Claims 17-18, 21-22, 24, 28, 30, and 35 have been considered on the merits, insofar as they read on the elected species of hydrolase, a muramidase of SEQ ID NO: 1, a bolus addition, a fed batch fermentation process wherein the at least one enzyme is added to the fermentation broth, adding up to 500 mg/kg of the at least one enzyme per kg of fermentation broth, and a Bacillus species. All arguments have been fully considered.
Withdrawn Rejections
Rejections under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, are withdrawn in view of applicant’s amendments.
Rejections of Claims 32 and 40 under 35 U.S.C. 103 as being unpatentable over Liu et al (WO 2018/113743 A1; 6/28/2018.) in view of Zhou et al (Molecules. 2018;23(1):1-14.) and Kimmenade et al (WO 2009/102755 A1; 8/20/2009.) are withdrawn in view of applicant’s amendments – Claims 32 and 40 have been canceled.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102 of this title, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
Claims 17-18, 21, 24, and 35 are rejected under 35 U.S.C. 103 as being unpatentable over Liu et al (WO 2018/113743 A1; 6/28/2018.) in view of Zhou et al (Molecules. 2018;23(1):1-14.) and Kimmenade et al (WO 2009/102755 A1; 8/20/2009.).
The instant claims recite a submerged fermentation process of producing a polypeptide of interest, the process comprising: a) fermenting a microorganism that produces the polypeptide of interest in a fermentation broth, and b) adding at least one enzyme to the fermentation broth, or co-expressing the at least one enzyme in the microorganism and secreting the at least one enzyme into the fermentation broth, in an amount sufficient to reduce the viscosity of the fermentation broth compared to when the at least one enzyme is not added or co-expressed.
Liu teaches a method of producing a polypeptide comprising cultivating a host cell under conditions conducive for production of the polypeptide, and recovering the polypeptide (p.61 line 24-27), wherein the polypeptide has lysozyme (lysozyme is a hydrolase, p.1 line 14) activity (p.23 line 15), the cells are cultivated by fermentation including fed-batch fermentation (p.61 line 30-31), the polypeptide is recovered from a fermentation broth (a liquid medium – a submerged fermentation) (p.62 line 10-11), the fermentation broth contains enzymes (p.63 line 27, p.64 line 16), and the host cell is Bacillus (p.59 line 1-5).
Liu does not teach the method comprises adding at least one enzyme to the fermentation broth, or co-expressing the at least one enzyme in the microorganism and secreting the at least one enzyme into the fermentation broth, in an amount sufficient to reduce the viscosity of the fermentation broth compared to when the at least one enzyme is not added or co-expressed (claim 17).
However, Liu does teach a fermentation process wherein polypeptides are recovered from a fermentation broth. Before the effective filing date of the claimed invention, it was well-known in the art that an increase in viscosity of fermentation broth leads to a reduction in mass transfer efficiency and makes oxygen transfer from a liquid phase to cells difficult (p.8 para 3 & 6), as evidenced by Zhou. In addition, Kimmenade teaches 2 µl lysozyme solution reduces broth viscosity (para 00129).
Thus, before the effective filing date of the claimed invention, it would have been obvious to one of ordinary skill in the art to add a sufficient amount of lysozyme (an enzyme) to a fermentation broth to reduce the viscosity of the fermentation broth, since Liu discloses a fermentation process wherein polypeptides are recovered from a fermentation broth, Zhou discloses that increased broth viscosity leads to a reduction in mass transfer efficiency and makes oxygen transfer from a liquid phase to cells difficult, and Kimmenade discloses that lysozyme reduces broth viscosity. Moreover, before the effective filing date of the claimed invention, one of ordinary skill in the art would have been motivated by the cited reference and routine practice to add a sufficient amount of lysozyme (an enzyme) to a fermentation broth with a reasonable expectation for successfully producing a polypeptide.
Claim 22 is rejected under 35 U.S.C. 103 as being unpatentable over Liu et al (WO 2018/113743 A1; 6/28/2018.) in view of Zhou et al (Molecules. 2018;23(1):1-14.) and Kimmenade et al (WO 2009/102755 A1; 8/20/2009.) as applied to claims 17-18, 21, 24, and 35 above, further in view of Lindskog et al (Biopharmaceutical Processing Development, Design, and Implementation of Manufacturing Processes. 2018, Chapter 31, p.625-635.).
References cited above do not teach the method wherein the at least one enzyme is added as a bolus addition (claim 22).
However, Liu does teach a fermentation process including fed-batch fermentation. Lindskog teaches fed-batch fermentation wherein a feed solution can be added intermittently (bolus feed), and bolus feed could be a better option when in very large-scale simplicity and robustness might need to be prioritized before sophistication (p.628 para 4).
Thus, before the effective filing date of the claimed invention, it would have been obvious to one of ordinary skill in the art to add enzyme as a bolus addition, since Liu discloses a fermentation process including fed-batch fermentation, and Lindskog discloses a feed solution can be added as a bolus feed, and bolus feed could be a better option in certain situations. Moreover, before the effective filing date of the claimed invention, one of ordinary skill in the art would have been motivated by the cited reference and routine practice to add enzyme as a bolus addition with a reasonable expectation for successfully producing a polypeptide.
Claim 28 is rejected under 35 U.S.C. 103 as being unpatentable over Liu et al (WO 2018/113743 A1; 6/28/2018.) in view of Zhou et al (Molecules. 2018;23(1):1-14.) and Kimmenade et al (WO 2009/102755 A1; 8/20/2009.) as applied to claims 17-18, 21, 24, and 35 above, further in view of Schnorr et al (WO 2013/076253 A1; 5/30/2013.).
References cited above do not teach the method wherein the at least one enzyme comprises a GH24 or GH25 muramidase comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence shown in any of SEQ ID NO: 1 (claim 28).
However, Liu does teach a fermentation process wherein polypeptides are recovered from a fermentation broth, Zhou does teach that increased broth viscosity leads to a reduction in mass transfer efficiency and makes oxygen transfer from a liquid phase to cells difficult, and Kimmenade does teach that lysozyme reduces broth viscosity. Schnorr teaches a GH25 lysozyme having SEQ ID NO: 4 (the claimed SEQ ID NO: 1, see Result 3 in Search results filed on 07/18/2025) may be used in a fermentation process to control microbial growth (p.47 last para).
Thus, before the effective filing date of the claimed invention, it would have been obvious to one of ordinary skill in the art to incorporate the claimed GH25 muramidase, since Liu discloses a fermentation process, and Schnorr discloses that the claimed GH25 muramidase may be used in a fermentation process to control microbial growth. Moreover, before the effective filing date of the claimed invention, one of ordinary skill in the art would have been motivated by the cited reference and routine practice to incorporate the claimed GH25 muramidase with a reasonable expectation for successfully producing a polypeptide.
Claim 30 is rejected under 35 U.S.C. 103 as being unpatentable over Liu et al (WO 2018/113743 A1; 6/28/2018.) in view of Zhou et al (Molecules. 2018;23(1):1-14.) and Kimmenade et al (WO 2009/102755 A1; 8/20/2009.) as applied to claims 17-18, 21, 24, and 35 above, further in view of Robinson (Essays Biochem. 2015;59:1-41.).
References cited above do not teach the method comprises adding up to 500 mg/kg of the at least one enzyme per kg of fermentation broth (claim 30).
Robinson teaches the amount of enzyme controls reaction rates, the relationship between the reaction rate and the amount of enzyme is a simple linear relationship, and substrate concentration effects the linear relationship (p.10 para 3-5).
Thus, before the effective filing date of the claimed invention, it would have been obvious to one of ordinary skill in the art to incorporate an optimized concentration of an enzyme, since it was well-known in the art that the amount of enzyme controls reaction rates, and that substrate concentration effects the amount of enzyme used in the reaction. In other words, the optimized amount of enzyme used in a fermentation broth depends on other parameters such as substrate concentration. Moreover, before the effective filing date of the claimed invention, one of ordinary skill in the art would have been motivated by the cited reference to incorporate an optimized concentration of an enzyme with a reasonable expectation for successfully producing a polypeptide.
Response to Arguments
Applicant argues that Kimmenade teaches away from the Applicant's invention, and that Kimmenade discloses polypeptides "that possess antimicrobial activity" (Abstract), as such, the skilled person would not consider adding these polypeptides to a fermentation process with an expectation of improving the yield.
These arguments are not found persuasive because applicant’s arguments regarding Kimmenade’s teaching of polypeptides appear to have no connection to the subject at issue, because Kimmenade is relied upon to teach that lysozyme solution reduces broth viscosity.
Applicant argues that Lindskog fails to cure the defects of Liu, Zhou and Kimmenade.
These arguments are not found persuasive because Lindskog is relied upon to teach that the at least one enzyme is added as a bolus addition.
Applicant argues that Schnorr fails to cure the defects of Liu, Zhou and Kimmenade.
These arguments are not found persuasive because Schnorr is relied upon to teach that the at least one enzyme comprises a GH24 or GH25 muramidase comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence shown in any of SEQ ID NO: 1.
Applicant argues that Robinson fails to cure the defects of Liu, Zhou and Kimmenade.
These arguments are not found persuasive because Robinson is relied upon to teach that the method comprises adding up to 500 mg/kg of the at least one enzyme per kg of fermentation broth.
Conclusion
No claims are allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any extension fee pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Contact Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to LYNN Y FAN whose telephone number is (571)270-3541. The examiner can normally be reached on M-F 7am-4pm.
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/Lynn Y Fan/
Primary Examiner, Art Unit 1759