DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Election/Restrictions
Applicant’s election without traverse of claims 1-5, 8-10, 12, 14, 16, 19, 33, 40, 47, and 49; as well as the species elections of option (a) in claim 4; option (a) in claim 19(iii); option a in claim 33(iii); and option (a) in claim 40(v) in the reply filed on 9/11/2025 is acknowledged.
Claims 58, 60, and 62-63 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 9/11/2025.
In view of the prior art search, the species election of a single amino acid modification as recited in claim 4, options (a)-(d), is withdrawn.
Priority
This application is a 371 of PCT/US21/52938 (9/30/2021) which claims benefit of 63/085,699 (9/30/2020) as reflected in the filing receipt issued on 11/29/2023.
Information Disclosure Statement
The information disclosure statement (IDS) filed on 8/1/2023 is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Drawings
The drawings are objected to for the following reasons:
37 CFR 1.84(u)(1) states “View numbers must be preceded by the abbreviation “FIG.”.
In the current case, the view numbers for Figures 1-33 are preceded by the word “Figure” instead of the abbreviation “FIG.”. View numbers should be updated to recite the abbreviation “FIG.”. Any changes to the drawings should also be reflected in the specification.
Corrected drawing sheets in compliance with 37 CFR 1.121(d) are required in reply to the Office action to avoid abandonment of the application. Any amended replacement drawing sheet should include all of the figures appearing on the immediate prior version of the sheet, even if only one figure is being amended. The figure or figure number of an amended drawing should not be labeled as “amended.” If a drawing figure is to be canceled, the appropriate figure must be removed from the replacement sheet, and where necessary, the remaining figures must be renumbered and appropriate changes made to the brief description of the several views of the drawings for consistency. Additional replacement sheets may be necessary to show the renumbering of the remaining figures. Each drawing sheet submitted after the filing date of an application must be labeled in the top margin as either “Replacement Sheet” or “New Sheet” pursuant to 37 CFR 1.121(d). If the changes are not accepted by the examiner, the applicant will be notified and informed of any required corrective action in the next Office action. The objection to the drawings will not be held in abeyance.
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 2-5, 8-9, 19, 33, and 40 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
Regarding written description, 35 U.S.C. 112(a) and the first paragraph of pre-AlA 35 U.S.C. 112 require that the "specification shall contain a written description of the invention ...." This requirement is separate and distinct from the enablement requirement. Ariad Pharm., Inc. v. Eli Lilly & Co., 598 F.3d 1336, 1340, 94 USPQ2d 1161, 1167 (Fed. Cir. 2010). To satisfy the written description requirement, a patent specification must describe the claimed invention in sufficient detail that one skilled in the art can reasonably conclude that the inventor had possession of the claimed invention (MPEP § 2163(I)).
MPEP 2163(II)(A)(3)(a)(i and ii) states that the written description requirement for a claimed genus may be satisfied through sufficient description of a representative number of species by actual reduction to practice, reduction to drawings, or by disclosure of relevant, identifying characteristics, i.e., structure or other physical and/or chemical properties, by functional characteristics coupled with a known or disclosed correlation between function and structure, or by a combination of such identifying characteristics, sufficient to show the applicant was in possession of the claimed genus.
A "representative number of species" means that the species which are adequately described are representative of the entire genus. See AbbVie Deutschland GmbH & Co., KG v. Janssen Biotech, Inc., .759 F.3d 1285, 1300, 111 USPQ2d 1780, 1790 (Fed. Cir. 2014). Thus, when there is substantial variation within the genus, one must describe a sufficient variety of species to reflect the variation within the genus.
In the instant case, claim 2 recites “the engineered lactate oxidoreductase of claim 1, comprising a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 1-16, provided at least one of the amino acids at a position in said sequence corresponding to positions 10 to 30, positions 119-139, positions 154-174, or positions 175 to 195 of SEQ ID NO: 1 is different from the amino acid occupying the corresponding position in said SEQ ID NO: 1-16”. Claim 3 recites “the engineered lactate oxidoreductase of claim 2, comprising a sequence
having at least 90% sequence identity to SEQ ID NO: 1 provided at least one of positions 10 to
30 or positions 175 to 195 is different from the amino acid occupying the corresponding position
in SEQ ID NO: 1”.
There is not sufficient written description support for a lactate oxidoreductase enzyme having 90% sequence identity to SEQ ID NOs: 1-16 that retains the claimed function, i.e. lactate oxidoreductase activity and increased stability as recited in claim 1. There is not a disclosed structure-function relationship for the claimed sequence. 90% identity means that 10% of the amino acids can vary from the original sequence, yielding a large number of potential variant sequences for each of SEQ ID NOs: 1-16. A skilled artisan would not be aware of which of these potential variants of SEQ ID NOs: 1-16 are within the scope of the claim. The claimed lactate oxidoreductases are directed to some specific amino acid substitutions, i.e. in positions 10 to 30, positions 119-139, positions 154-174, or positions 175 to 195 of SEQ ID NO: 1. However, an additional 10% of residues aside from these specified residues may vary, and it is unclear which of these variants would result in a lactate oxidoreductase having the claimed function. There are no disclosed examples of representative sequences with modifications other than substitutions at the positions specifically disclosed in the claims, or any disclosure of regions or residues where variation may or may not occur, to provide a sufficient structure-function relationship for a lactate oxidoreductase enzyme with 90% identity to SEQ ID NO: 1.
It is established in the art that substitutions of even a single amino acid residue can have large impacts on function, and the effect of these substitutions further depends on whether the substitution is conservative, and therefore expected to maintain the overall enzyme structure, or non-conservative (see Minagawa et al. pg. 78-79 “Results and discussion” and Table 1 demonstrating the varying effects on enzyme function for mutations at different residues in lactate oxidase; and Leitgeb et al. pg. 5 para. 46-47 for discussion on conservative mutations). Thus, substitutions of enzyme amino acid residues are considered unpredictable and a skilled artisan would not be aware of which 10% of amino acids could be substituted in the lactate oxidoreductase of the invention and maintain the claimed function.
For these reasons, claims 2 and 3 fail to comply with the written description requirement, as the disclosure does not reasonably convey to a person having ordinary skill in the art that the inventors had possession of the entire scope of the claimed invention at the time of filing.
Claims 4-5, 8-9, 19, 33, and 40 are included in the rejection because they depend on a rejected claim and do not clarify the issue.
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 8-10, 12, 14, 16, 19, 33, 40, and 49 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.
Claim 8 recites the limitation “wherein the modification includes a substitution of the wild-type amino acid residue with an amino acid residue Cys” (emphasis added). Similarly, claims 9, 10, 12, 14, 19, 33, 40, and 49 recite “wherein the modification includes”; and claim 16 recites “which includes”. The use of the term “includes” makes it unclear whether these limitations are required or optional. As an example, it is not clear whether the limitation in claim 8 means “wherein the modification is”, in which case the substitution of the WT residue at positions 20, 185, 129, or 164 must be to a Cys residue; or if it means “wherein the modification could be”, meaning that the specific substitution of a Cys residue at positions 20, 185, 129, or 164 is one of multiple options, and therefore it is not required that the WT residue at these positions be substituted with a Cys residue. If it is intended that these limitations are required, it is suggested that the claims be amended to clarify the wording, for example, “wherein the modification is” rather than “wherein the modification includes”.
For the purposes of examination, the term “includes” as used in claims 8-10, 12, 14, 16, 19, 33, 40, and 49 is interpreted to mean that the recited limitation is one of multiple potential options, and therefore is not required. In the case of claims 8-10, 12, 14, 19, 33, 40, and 49, this means that the oxidoreductase is required to have a substitution at the claimed position, but a substitution of any amino acid at this position reads on 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)(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.
Claims 1-3, 12, 14, and 49 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Leitgeb et al., US 2013/0071868 A1.
Regarding claim 1, Leitgeb teaches engineered lactate oxidases with increased stability compared to wild-type (Leitgeb pg. 1 para. 2). The instant specification, pg. 7 para. 58, states that engineered stable lactate oxidoreductases include lactate oxidases.
Regarding claim 2, Leitgeb teaches that the engineered lactate oxidase with increased stability has a sequence according to SEQ ID NO: 2 (Leitgeb pg. 2 para. 20). SEQ ID NO: 2 of Leitgeb is 99.5% identical to instant SEQ ID NO: 1 (see sequence alignment in OA Appendix). Leitgeb teaches that the mutant lactate oxidase has a substitution at position Tyr191 and at least one further amino acid substitution at a position selected from the group consisting of Gly36, Ala95, Thr103, Glu160, Val198, Asn212, Ala/Gly232 and Phe277 of SEQ ID NO: 2 (Leitgeb pg. 4 para. 32-33). Thus, Leitgeb teaches an engineered lactate oxidoreductase wherein an amino acid in positions 154-174 (Glu160) or 175-195 (Tyr191) is different from the corresponding WT position.
Regarding claim 3, Leitgeb teaches an engineered lactate oxidoreductase with at least 90% identity to instant SEQ ID NO: 1 (Leitgeb SEQ ID NO: 2, see sequence alignment in OA Appendix) wherein an amino acid in positions 175-195 (Tyr191) is different from the corresponding WT position (Leitgeb pg. 4 para. 32-33).
Regarding claim 12, Leitgeb teaches an engineered lactate oxidoreductase with a modification of the WT residue at position 212 of instant SEQ ID NO: 1, specifically Asn212Glu and Asn212Asp (Leitgeb pg. 5 para. 51-52). As discussed in the rejection under 35 U.S.C. § 112(b) above, the use of the term “includes” makes it unclear whether the limitation “wherein the modification includes a substitution of the wild-type amino acid residue with an amino acid residue Lys” is a required or optional claim element. As set forth above, for the purposes of examination this is being interpreted to mean that the specific substitution of the WT residue for Lys at positions 212 is optional. Therefore, the teachings of Leitgeb, directed to an engineered lactate oxidoreductase comprising a modification at position 212 of instant SEQ ID NO: 1, anticipate claim 12.
Regarding claims 14 and 49, Leitgeb teaches an engineered lactate oxidoreductase with a modification of the WT residue at position 95 of instant SEQ ID NO: 1 to Ser, Ala95Ser (Leitgeb pg. 5 para. 52).
Claims 1-5 and 8-9 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Minagawa et al., Cellular and molecular life sciences; 64(1):77-81.
Regarding claim 1, Minagawa teaches engineered lactate oxidases with increased stability compared to wild-type (Minagawa “Abstract”).
Regarding claim 2, Minagawa teaches lactate oxidase (LOX) from Aerococcus viridans (Minagawa “Abstract”). The lactate oxidoreductase according to instant SEQ ID NO: 1 is 100% identical to Aerococcus viridans lactate oxidase (see sequence alignment in OA Appendix). Therefore, the lactate oxidase taught by Minagawa is expected to have an identical sequence to instant SEQ ID NO: 1. Minagawa teaches mutations at positions 160, 198, 103, 345, 164, 355, 104, 232, 36, 75, 185, 225, and 277 (Minagawa pg. 78 Table 1). Thus, Minagawa teaches an engineered lactate oxidoreductase according to SEQ ID NO: 1, wherein an amino acid in positions 154-174 (D164) and 175-195 (V185) is different from the amino acid occupying the position in the WT sequence according to SEQ ID NO: 1.
Regarding claim 3, Minagawa teaches an engineered lactate oxidoreductase according to SEQ ID NO: 1, wherein an amino acid in positions 175-195 (V185) differs from the amino acid occupying the position in the WT sequence according to SEQ ID NO: 1 (Minagawa Table 1).
Regarding claim 4, Minagawa teaches an engineered lactate oxidoreductase comprising a modification at a position corresponding to positions 185 and 164 of the amino acid sequence set forth in SEQ ID NO: 1 (Minagawa pg. 78 Table 1).
Regarding claim 5, Minagawa teaches an engineered lactate oxidoreductase comprising a modification at a position corresponding to position 185 of the amino acid sequence set forth in SEQ ID NO: 1 (Minagawa pg. 78 Table 1).
Regarding claims 8 and 9, as discussed in the rejection under 35 U.S.C. § 112(b) above, the use of the term “includes” makes it unclear whether the limitation “wherein the modification includes a substitution of the wild-type amino acid residue with an amino acid residue Cys” is a required or optional claim element. As set forth above, for the purposes of examination this is being interpreted to mean that the specific substitution of the WT residue for Cys at positions 20, 185, 129, and 164 is optional. Therefore, the teachings of Minagawa, directed to an engineered lactate oxidoreductase comprising a modification at positions 185 and 164 of instant SEQ ID NO: 1, anticipate claims 8 and 9.
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.
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.
Claims 10 and 16 are rejected under 35 U.S.C. 103 as being unpatentable over Leitgeb as applied to claims 1-3, 12, 14, and 49 above, and further in view of Hiraka et al., Biosensors and Bioelectronics; 103:163-70.
Leitgeb teaches an engineered lactate oxidase according to claim 1, as set forth above. Leitgeb does not teach that the engineered lactate oxidoreductase further comprises a modification at position 96 of SEQ ID NO: 1, wherein the modification includes a substitution of the WT residue to Leu, as recited in claim 10.
Regarding claim 10, Hiraka teaches an engineered lactate oxidase from Aerococcus viridans, wherein the lactate oxidase has a substitution at position 96, Ala96Leu (Hiraka “Abstract”). Hiraka teaches that the stability of Ala96Leu mutant was almost indistinguishable from the wild-type enzyme (Hiraka pg. 166 Section 3.2). Hiraka teaches that L-Lactate is an essential biomarker for clinical diagnostics, sports medicine, and food quality control (Hiraka pg. 163 para. 1). Hiraka teaches that the major biological component of L-lactate biosensors is L-lactate oxidase (LOx), which can be obtained from Aerococcus viridans, AvLOx (Hiraka pg. 164 para. 1). Developing lactate sensors with minimal oxygen interference remains challenging in the field of biomedical engineering research, and thus it is desirable to engineer an AvLOx that minimizes the effects of oxygen (Hiraka pg. 164 para. 1 and 4-5). Hiraka teaches that the Ala96Leu mutant of AvLOx decreases the effects of oxygen interference for biosensing purposes, and the Ala96Leu mutant has higher dehydrogenase activity with reduced oxidase activity (Hiraka pg. 180 “Conclusion”). Hiraka teaches that the Ala96Leu mutant has the potential to be a practical L-lactate biosensor without oxygen interference (Hiraka pg. 180 “Conclusion”).
Regarding claim 16, Leitgeb and Hiraka teach the lactate oxidoreductase of claim 10, which includes a mutation of Ala96Leu, as discussed above. Hiraka teaches that the Ala96Leu mutant has increased dehydrogenase activity with reduced oxidase activity compared to the WT (Hiraka pg. 180 “Conclusion”)
It would have been obvious to a skilled artisan, before the effective filing date, to combine the teachings of Leitgeb and Hiraka to create an engineered lactate oxidase with a sequence according to SEQ ID NO: 1 and a modification of Ala96Leu. Both Leitgeb and Hiraka teach engineered lactate oxidases from A. viridans. Leitgeb teaches amino acid substitutions which enhance the stability of the enzyme, including a lactate oxidase with multiple amino acid substitutions (Leitgeb pg. 5 para. 50-57). Hiraka similarly teaches amino acid substitutions of A. viridans lactate oxidase. It would have been obvious to a person having ordinary skill in the art that a substitution of Ala96Leu, as taught by Hiraka, could be incorporated in an engineered lactate oxidase as taught by Leitgeb.
A person of ordinary skill in the art would have been motivated to modify the lactate oxidase of Leitgeb and include a substitution of Ala96Leu because this substitution results in higher dehydrogenase activity with reduced oxidase activity, therefore decreasing the effects of oxygen interference for biosensing purposes (Hiraka pg. 180 “Conclusion”). Leitgeb teaches that the engineered lactate oxidase is used as a biosensor and it is of interest to increase stability of lactate oxidase in order to increase its shelf-life and use time as a biosensor (Leitgeb pg. 2 para. 17). Thus, as the invention of Leitgeb is directed to engineering lactate oxidases that are better biosensors, it would be considered advantageous to incorporate an Ala96Leu mutation as taught by Hiraka, as this mutation provides additional benefits for biosensing, specifically by decreasing the effects of oxygen interference. Leitgeb teaches a more stabilized lactate oxidase for biosensing, as a lack of stability is a drawback of using lactate oxidases in biosensors (Leitgeb pg. 2 para. 17). A skilled artisan would have been motivated to further improve upon this lactate oxidase with a mutation that alleviates another challenge associated with using lactate oxidases in biosensing, oxygen interference.
A skilled artisan would have a reasonable expectation of success in modifying the lactate oxidase taught by Leitgeb with the Ala96Leu modification taught by Hiraka to achieve the predictable outcome of an enzyme having greater stability and decreased oxygen interference. Leitgeb teaches a lactate oxidase with modifications to increase stability. Hiraka teaches that a substitution, Ala96Leu, results in decreased oxygen interference in the same enzyme. Thus, a skilled artisan could reasonably expect success in incorporating an Ala96Leu mutation in the enzyme taught by Leitgeb, as this substitution has been proven to result in beneficial alterations of function by Hiraka.
Claim 47 is rejected under 35 U.S.C. 103 as being unpatentable over Leitgeb as applied to claims 1-3, 12, 14, and 49 above, in view of Ma et al., ChemElectroChem; 6(4):958-75.
Leitgeb teaches an engineered lactate oxidase according to claim 1, as set forth above.
Leitgeb does not teach that the engineered lactate oxidoreductase further comprises a fused cytochrome domain of flavocytochrome b2, as recited in claim 47.
Regarding claim 47, Ma teaches that the direct electron transfer (DET) of enzymes has been utilized to develop biosensors to detect glucose, lactate, and other analytes, and that while some enzymes exhibit direct electron transfer between their active-site cofactor and an electrode, other oxidoreductases depend on acquired cytochrome domains as built-in redox mediators, which transfer electrons between the active-site cofactor and a redox partner protein (Ma “Abstract”). Ma teaches the use of cytochrome domains from flavocytochrome b2 for biosensing (Ma pg. 970 Section 6.2). Ma teaches that DET enzymes for biosensing have been developed by fusion of a biosensing oxidoreductases such as glucose dehydrogenase to a cytochrome domain to create new multi-cofactor enzymes capable of DET (Ma pg. 971 “Generating DET Enzymes by Fusion of a Cytochrome Domain”). Ma teaches that the fusion of cytochromes to catalytically interesting enzymes is a promising strategy to generate DET-enzymes for new substrates and analytes, and that initial research has proven the validity of this concept which can be used for rational design of fusion enzymes to obtain DET-capable biosensor catalysts (Ma pg. 973 para. 3).
It would have been obvious to a skilled artisan, before the effective filing date, to combine the teachings of these references and create an engineered lactate oxidoreductase comprising a fused cytochrome domain from flavocytochrome b2. Leitgeb teaches an engineered lactate oxidoreductase for use as a biosensor (Leitgeb pg. 2 para. 17). Ma teaches fusion proteins comprising biosensing enzymes fused to a cytochrome domain. It would have been obvious to one skilled in the art that the biosensing lactate oxidoreductase taught by Leitgeb could be fused to a cytochrome domain as taught by Ma.
A person of ordinary skill in the art would have been motivated to create a lactate oxidase fused to a cytochrome domain of flavocytochrome b2 because it is of interest to develop oxidoreductases capable of direct electron transfer, as the performance of oxidoreductase biosensors depends on the DET rate (Ma pg. 959 para. 1-3). As Ma teaches the fusion of cytochrome domains to biosensing oxidoreductases is a successful technique for developing DET enzymes for biosensing, a skilled artisan would be motivated to fuse a cytochrome domain to the oxidoreductase taught by Leitgeb as an improvement to the biosensing capability of the enzyme.
A skilled artisan would have a reasonable expectation of success in creating a lactate oxidase fused to a cytochrome domain from flavocytochrome b2 because Ma teaches that initial research has proven the validity of fusing of cytochromes to catalytically interesting enzymes to generate DET-enzymes, so a person having ordinary skill in the art could reasonably expect success in creating such a fusion enzyme with a known oxidoreductase biosensing enzyme such as the lactate oxidase taught by Leitgeb.
Allowable Subject Matter
The prior art does not teach an engineered lactate oxidoreductase having at least 90% sequence identity to any of SEQ ID NOs: 1-16 and also having a modification at a position corresponding to positions 20 or 129 of SEQ ID NO: 1, as required by claims 19, 33, and 40. The closest prior art is Leitgeb et al., US 2013/0071868 A1; Minagawa et al., Cellular and molecular life sciences; 64(1):77-81; and Hiraka et al., Biosensors and Bioelectronics; 103:163-70.
Leitgeb teaches engineered lactate oxidoreductases with amino acid substitutions at position Tyr191, Gly36, Ala95, Thr103, Glu160, Val198, Asn212, Ala/Gly232 and Phe277 corresponding to SEQ ID NO: 1. Leitgeb does not teach or suggest a modification at positions 20, 185, 129, or 164 as required by claims 19, 33, and 40. There is nothing in the teachings of Leitgeb to provide motivation for a skilled artisan to modify any of these residues in SEQ ID NO: 1, or corresponding residues in SEQ ID NOs: 2-16.
Minagawa teaches engineered lactate oxidases with modifications at a position corresponding to positions 185 and 164 of the amino acid sequence set forth in SEQ ID NO: 1 (Minagawa pg. 78 Table 1). Minagawa teaches the substitutions D164V and V185M (Minagawa Table 1). Minagawa does not teach substitutions at positions 20 or 129, and there is nothing in the teachings of Minagawa to provide motivation for a skilled artisan to modify any of these residues in SEQ ID NO: 1, or corresponding residues in SEQ ID NOs: 2-16.
Hiraka teaches an engineered lactate oxidase from Aerococcus viridans, with a sequence according to instant SEQ ID NO: 1, wherein the lactate oxidase has a substitution at position 96, Ala96Leu (Hiraka “Abstract”). Hiraka does not teach or suggest substitutions at positions 20, 185, 129, or 164 of SEQ ID NO: 1, or provide motivation for a skilled artisan to modify any of these residues in SEQ ID NO: 1 or corresponding residues in SEQ ID NOs: 2-16.
The prior art does not teach lactate oxidoreductases having at least 90% identity to any of SEQ ID NOs: 1-16 and a modification at the positions corresponding to positions 20 or 129 of SEQ ID NO: 1. The prior art does not teach a lactate oxidoreductase with substitutions at positions 20, 185, and 96 as required by claim 19; at positions 129, 164, and 96 as required by claim 33; or at positions 20, 185, 129, 164, and 96 as required by claim 40.
Claims 19, 33, and 40 would be allowable if rewritten to overcome the rejections under 35 U.S.C. 112(a) and (b) set forth in this Office action, and to include all of the limitations of the base claim and any intervening claims.
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to EMILY F EIX whose telephone number is (571)270-0808. The examiner can normally be reached M-F 8am-5pm ET.
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/EMILY F EIX/Examiner, Art Unit 1653
/JENNIFER M.H. TICHY/Primary Examiner, Art Unit 1653