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 .
DETAILED ACTION
Applicant filed a claim amendment on July 9, 2025. Claims 1-20, 25, 31-32, and 34-35 are canceled. Claims 21, 33, and 38 are amended. The previous objection to claim 38 and 112(b) rejection are withdrawn.
Claims 21-24, 26-30, 33, and 36-38 are pending. Claims 36-37 are withdrawn.
The instant application is a U.S. national phase of PCT/EP2019/053716, filed February 14, 2019.
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on July 9, 2025 has been entered.
Claim Objections
Claims 21, 33, and 38 are objected to because of the following informalities:
Claims 21 and 38, lines 1-5, needs to be changed to “A method for the purification of collagenase type I, collagenase type II, neutral protease and clostripain, from a mixture of substances ing
Claim 33 has an improper status identifier, (Currently Amended), while the claim is not amended. The status needs to be changed to (Previously Presented).
Appropriate correction is required.
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.
Claim 23 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.
Claim 23 recites the limitation "the at least one enzyme" in line 2. There is insufficient antecedent basis for this limitation in the claim.
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 21-24, 26-30, 33, and 38 are rejected under 35 U.S.C. 103 as being unpatentable over JP 2004535197 A (hereinafter “JP”) in view of Kula et al. (Biochem and Biophys Research Comm. 1976. Vol. 69, No. 2, pgs. 389- 396, previously cited in PTO-892 mailed 10/17/2024, hereinafter “Kula”) and Fres et al. (Journal of Lipid Research Volume 51, 2010, pgs. 2454-2459, hereinafter “Fres”).
JP teaches a method for partially or completely purifying at least one kind of enzyme contained in an excess fermentation product of Clostridium histolyticum (abstract). JP teaches the microorganism Clostridium histolyticum extracellularly produces a complex enzyme mixture containing collagenase, various proteinaceous enzymes and low molecular weight components when cultured in a peptone-containing nutrient medium, wherein the main components are type I and type II collagenases, clostripain, and a neutral protease [0002]. JP teaches the enzyme of the Clostridium histolyticum overferment is separated by a single-stage or preferably a multi-stage chromatographic method exclusively using chromatographic materials based on styrene / divinylbenzene and / or especially on the hydroxylapatite ceramics, which results in a very rapid and cost-effective enzyme purification [0007]. JP teaches hydroxylapatite is a calcium phosphate represented by the general formula Ca10 (PO4 )6 (OH)2 that allows protein binding for ionic electrostatic exchange and strong ionic complex salt binding [0007]. JP teaches three elution steps, Fraction 1 obtained in the first elution stage contains exclusively low molecular weight components. Fraction 2 of the second elution stage also contains a neutral protease (caseinase) in addition to low molecular weight components. Fraction 3 of the third elution step also contains low molecular weight components, including all clostripains and all type I and type II collagenases [0017]. Fraction 3 is subjected to a second chromatography step to further separate the clostripains and collagenases into further fractions [0018]. JP teaches the method does not have any protein precipitation steps that can cause unwanted structural changes of the protein, such as ammonium sulfate [0011].
JP does not teach the method utilizes hydrophobic interaction chromatography comprising material propylene glycol or butyl sepharose, nor the type of salt used in the aqueous solution in the mobile phase, such as ammonium sulfate or potassium chloride. JP also does not teach the clostripain and collagenases are further separated in the single chromatography step.
However, Kula teaches a purification method of collagenase A and clostripain from Clostridium histolyticum (title). Kula teaches the use of hydrophobic chromatography (HIC) utilizing butyl Sepharose to separate clostripain in a homologous series of hydrocarbon-coated agaroses for the development of fractionators which meets the limitation of claim 21 (abstract). Kula teaches clostripain was extracted by passing the culture filtrate though the butyl agarose, and then collagenase A was extracted by passing the excluded mixture through heptylagarose (abstract). Kula teaches the enzymes were a lyophilized collagenase preparation containing several extracellular proteins from Clostridium histolyticum served as a source for both collagenase and clostripain (pg. 390, para 2). Kula teaches separation in a step elution scheme by passage of the crude protein first on the butyl sepharose to extract the clostripain, and then by applying the excluded proteins on the heptyl agarose in order to purify the collagenase (pg. 392, para 3).
JP and Kula do not teach butyl sepharose that is a cross-linked agarose with 3-n-butoxy-2-hydroxypropyl residues.
However, Fres teaches optimizing separation and purification of a recombinant farnesylated guanylate-binding protein 1 (hGBP1), a dynamin-related large GTPase, using Butyl Sepharose High Performance (HP) (abstract, pg. 2455, col. 2, para 1). As disclosed in the specification, the claimed method utilizes butyl Sepharose HP which is a cross-linked agarose with 3-n-butoxy-2-hydroxypropyl residues (pg. 5, para 2). Fres teaches baseline purification of the farnesylated protein was made possible by using the new HIC resin, Butyl Sepharose HP, which separation of the farnesylated and unmodified hGBP1 had not been possible with the previously available butyl Sepharose 4 Fast Flow (pg. 2456, col. 1, para 4-5). Fres also teaches the elution profile of hGBP1 from a preparative HIC using Butyl Sepharose High Performance (GE Healthcare) with an ammonium sulfate gradient from 0.5 M to 0 M, which meets the limitations recited in claims 21-23, 27-29, and 38 (pg. 2456, Fig. 1).
Therefore, it would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the JP method of separating collagenase type I & II, clostripain, and neutral protease from a C. histolyticum supernatant into separate fractions using a single chromatography step, as taught by JP, by replacing the chromatography materials with the newly available butyl Sepharose HP, which is a cross-linked agarose with 3-n-butoxy-2-hydroxypropyl residues as the stationary phase in the HIC, as taught by Fres. One of ordinary skill in the art would have been motivated to substitute chromatography materials because Fres discloses the more effective separation of closely related proteins utilizing butyl Sepharose HP. There would have been a reasonable expectation of success because Kula discloses separation of clostripain from collagenase in a step elution scheme using butyl sepharose.
Further, it would have been prima facie obvious to optimize the method of separating the aforementioned enzymes utilizing the HIC by incorporating a gradient or step elution scheme as taught by the prior art. One of ordinary skill in the art would have been motivated to separate the clostripain from collagenase fractions in the method taught by JP, as both are useful due to their narrow specificity, and thus depends on their being free of other proteolytic activities as taught by Kula (pg. 390, para 1). Thus, the method of purifying these four enzymes as described in JP could have been easily conceived by a person skilled in the art based on the technical matter described in Kula and Fres, wherein one of ordinary skill in the art would optimize the appropriate elution scheme based on well-known chromatography techniques in the art.
Response to Arguments
Applicant’s arguments with respect to the 103 rejection of claims 21-24, 26-30, 33, and 38 have been considered but are moot because the new ground of rejection does not rely on any reference applied in the prior rejection of record for any teaching or matter specifically challenged in the argument.
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to JESSICA EDWARDS whose telephone number is (571)270-0938. The examiner can normally be reached M-F 8am-5pm EST.
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/LOUISE W HUMPHREY/Supervisory Patent Examiner, Art Unit 1657
/JESSICA EDWARDS/
Examiner, Art Unit 1657