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 .
Claims 1-20 are pending and under consideration in this Office Action.
Claim Rejections - 35 USC § 112(b) or 35 U.S.C. 112 (pre-AIA ) 2nd Paragraph
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 1-20 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 pre-AIA the applicant regards as the invention.
Claim 1 recites the phrase “comprising using metal chelate affinity chromatography to purify tetanus toxin or variants thereof, wherein the tetanus toxin does not contain an affinity tag” which renders the claim vague and indefinite since the specific method steps for purifying tetanus toxin or variants thereof are not known and not stated in the claim. Dependent claims 2-20 are also rejected because they do not correct the defect.
Claim 3 recites the phrase “he tetanus toxin comprises an amino acid sequence as shown in SEQ ID NO: 1 or SEQ ID NO: 3” which renders the claim vague and indefinite since it is unclear if the claim is limited to the full length amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3, or any part or portion of SEQ ID NO: 1 or SEQ ID NO: 3. For examination purposes the claims will not be limited to any specific amino acid sequence.
Claim 4 recites the phrase “the variants have the following amino acid residue differences compared to SEQ ID NO: 1: C869S/A or C1093S/A” which renders the claim vague and indefinite since it is unclear if the variants comprise SEQ ID NO: 1 or SEQ ID NO: 3 and have the substitutions at the specific positions. Further claims 5 is vague and indefinite since it is unclear if the variants comprise SEQ ID NO: 1 or SEQ ID NO: 3 and have the substitutions at the specific positions.
Claim Rejections - 35 USC § 102
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 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.
Claim 1 is rejected under 35 U.S.C. 102(a)(1) as being anticipated by
CN116731130A (2023-09-12; PTO 892).
CN116731130A teaches a purification method of tetanus toxin or a variant thereof where the purification method of the tetanus toxin or the variant thereof uses a metal chelating affinity chromatography column for purification, and the tetanus toxin does not contain an affinity tag (see entire publication and abstract especially claims 1-10 of English translation). Thus, the reference teachings anticipate the claimed 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 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 1-20 are rejected under 35 U.S.C. 103 as being unpatentable over Rossetto et al. (Biochem J. 1992 Jul 1; 285:9-12; PTO 892) in view of Accession C4PD05 (07-JUL-2009; PTO 892), Accession P04958 (13-AUG-1987; PTO 892), Bornscheuer et al. (Curr Protoc Protein Sci. 2011 Nov;Chapter 26:Unit26.7; PTO 892), US20130053551(2013-02-28; PTO 781), CN116731130A (2023-09-12; PTO 892).
Rossetto et al. teach that tetanus toxin contains 14 histidine residues where six of them are localized in the light chain (L), one is present in the N-terminal half of the heavy chain (HN), and the remaining seven histidines are localized in the C-terminal half of the heavy chain
(Hc). Rossetto et al. teach using immobilized-metal-ion affinity chromatography with Chelating Superose-Zn(II) that histidines of Hc are exposed to the protein surface and are responsible for the binding of tetanus toxin and of Hc to the immobilized metal. Rossetto et al. teach that chromatography on Superose-Zn(II) is shown to be a simple and efficient method for the rapid isolation of tetanus toxin and of its Hc fragment, which can be extended to the botulinum neurotoxins. Rossetto et al. teach simple and effective methods for purification of tetanus toxin and fragments comprising using Superose-Zn(II) metal chelate affinity chromatography to the purify tetanus toxin and fragments. See entire publication and abstract especially EXPERIMENTAL section, Figs. 1-4, and pages 10-12
The teachings of the reference differ from the claims in that the reference does not teach the claimed method for purifying tetanus toxin or variants thereot
Accession C4PD05 teaches the tetanus toxin from Clostridium tetani having an amino acid sequence that is 100% identical to SEQ ID NO: 1 (see attached reference).
Accession P04958 teaches the tetanus toxin from Clostridium tetani having an amino acid sequence that is 100% identical to SEQ ID NO: 3 (see attached reference).
Bornscheuer et al. (Curr Protoc Protein Sci. 2011 Nov;Chapter 26:Unit26.7; PTO 892) teach protein engineering strategies to improve or change the properties of proteins, teach concepts for protein engineering using rational design including substitution and/or deletion of amino acids, directed evolution, and combinations of them where different strategies are presented for identifying the best mutagenesis method, how to identify desired variants by screening or selection, and examples for successful applications are shown which enable researchers to choose the most promising tools to solve their protein engineering challenges (see entire publication especially pages 26.7.1- 26.7.10 and Tables 26.7.1, 26.7.2, and 26.7.3).
US20130053551 teaches cation and anion exchange chromatography methods comprising the steps of i) applying a solution comprising the polypeptide to an ion exchange chromatography material, and ii) recovering the polypeptide with a solution comprising a denaturant and thereby purifying the polypeptide, whereby the ion exchange chromatography material comprises a matrix of cross-linked poly (styrene-divinylbenzene) to which ionic ligands have been attached, and wherein the solution comprising the polypeptide applied to the ion exchange chromatography material is free of the denaturant and the polypeptide adsorbed to the ion exchange chromatography material is recovered with a solution comprising a denaturant at a constant conductivity (see entire publication and claims especially paragraphs [0007]- [0041]. US20130053551 teaches the following in the claims:
1. A method for purifying a polypeptide comprising the step of:
recovering the polypeptide with a solution comprising a denaturant from an ion exchange chromatography material at constant conductivity and thereby purifying the polypeptide,
whereby the ion exchange chromatography material comprises a matrix of cross-linked poly (styrene-divinylbenzene) to which ionic ligands have been attached.
2. A method for producing a polypeptide comprising the following steps:
cultivating a prokaryotic or eukaryotic cell comprising a nucleic acid encoding the polypeptide,
recovering the polypeptide from the cells or/and the cultivation medium,
purifying the polypeptide with an ion exchange chromatography method comprising the following step
recovering the polypeptide with a solution comprising a denaturant from the ion exchange chromatography material and thereby producing a polypeptide,
whereby the ion exchange chromatography material comprises a matrix of cross-linked poly (styrene-divinylbenzene) to which ionic ligands have been attached.
3. The method of claim 1, characterized in that the denaturant is one denaturant and is selected from the group comprising urea, guanidine, urea-derivatives, and guanidine-derivatives.
4. The method of claim 2, characterized in that the denaturant is one denaturant and is selected from the group comprising urea, guanidine, urea-derivatives, and guanidine-derivatives.
5. The method of claim 1, characterized in that the ion exchange chromatography material is a cation exchange chromatography material.
6. The method of claim 2, characterized in that the ion exchange chromatography material is a cation exchange chromatography material.
7. The method of claim 5, characterized in that the ionic ligand is a sulfopropyl ligand or a carboxymethyl ligand.
8. The method of claim 6, characterized in that the ionic ligand is a sulfopropyl ligand or a carboxymethyl ligand.
9. The method of claim 1, characterized in that the ion exchange chromatography material is an anion exchange chromatography material.
10. The method of claim 2, characterized in that the ion exchange chromatography material is an anion exchange chromatography material.
11. The method of claim 9, characterized in that the ionic ligand is an ethyleneimine ligand or a quaternized ligand.
12. The method of claim 10, characterized in that the ionic ligand is an ethyleneimine ligand or a quaternized ligand.
13. The method of claim 1, further characterized in comprising the following steps:
wherein recovering the polypeptide from the ion exchange chromatography comprises a first ion exchange chromatography material by applying a solution comprising a denaturant,
applying the recovered polypeptide to a second ion exchange chromatography material, and
recovering the polypeptide from the second ion exchange chromatography material by applying a solution comprising a denaturant and thereby obtaining or purifying the polypeptide.
14. The method of claim 2, further characterized in comprising the following steps:
wherein recovering the polypeptide from the ion exchange chromatography comprises a first ion exchange chromatography material by applying a solution comprising a denaturant,
applying the recovered polypeptide to a second ion exchange chromatography material, and
recovering the polypeptide from the second ion exchange chromatography material by applying a solution comprising a denaturant and thereby obtaining or purifying the polypeptide.
15. The method according to claim 13, characterized in that
the first ion exchange chromatography material is an anion exchange chromatography material and the second ion exchange chromatography material is a cation exchange chromatography material, or
the first ion exchange chromatography material is a cation exchange chromatography material and the second ion exchange chromatography material is an anion exchange chromatography material.
16. The method according to claim 14, characterized in that
the first ion exchange chromatography material is an anion exchange chromatography material and the second ion exchange chromatography material is a cation exchange chromatography material, or
the first ion exchange chromatography material is a cation exchange chromatography material and the second ion exchange chromatography material is an anion exchange chromatography material.
17. The method of claim 13, characterized in that the recovered polypeptide is refolded prior to the applying to the second ion exchange chromatography material.
18. The method of claim 14, characterized in that the recovered polypeptide is refolded prior to the applying to the second ion exchange chromatography material.
19. The method of claim 1, wherein the purifying further comprises the following steps:
applying a first solution to the ion exchange chromatography material,
applying a second solution comprising the polypeptide to the ion exchange chromatography material, and
recovering and thereby producing or purifying the polypeptide with a fourth solution comprising a denaturant,
whereby the first solution comprises a first buffer substance, the second solution comprises a second buffer substance, and the fourth solution comprises a fourth buffer substance, wherein the fourth buffer substance comprises a denaturant.
20. The method of claim 2, wherein the purifying further comprises the following steps:
applying a first solution to the ion exchange chromatography material,
applying a second solution comprising the polypeptide to the ion exchange chromatography material, and
recovering and thereby producing or purifying the polypeptide with a fourth solution comprising a denaturant,
whereby the first solution comprises a first buffer substance, the second solution comprises a second buffer substance, and the fourth solution comprises a fourth buffer substance, wherein the fourth buffer substance comprises a denaturant.
21. The method according to claim 19, characterized in that after applying the second solution and prior to applying the fourth solution the following step is added:
applying a third solution to the ion exchange chromatography material,
whereby the third solution comprises a third buffer substance.
22. The method of claim 1, characterized in that the polypeptide has an amino acid sequence selected from the group of amino acid sequences of SEQ ID NO: 01, SEQ ID NO: 02, SEQ ID NO: 03, and SEQ ID NO: 04.
23. The method of claim 2, characterized in that the polypeptide has an amino acid sequence selected from the group of amino acid sequences of SEQ ID NO: 01, SEQ ID NO: 02, SEQ ID NO: 03, and SEQ ID NO: 04.
24. The method of claim 1, wherein the denaturant is a urea or a urea-derivate.
25. The method of claim 2, wherein the denaturant is a urea or a urea-derivate.
CN116731130A teaches a purification method of tetanus toxin or a variant thereof where the purification method of the tetanus toxin or the variant thereof uses a metal chelating affinity chromatography column for purification, and the tetanus toxin does not contain an affinity tag (see entire publication and abstract especially claims 1-10 of English translation).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify and/or combine the reference teachings to make the claimed invention by using the protein engineering strategies engineering strategies of Bornscheuer et al. on the tetanus toxin of Accession C4PD05 or Accession P04958 to make the variant, using the method of Rossetto et al. in combination with methods of US20130053551 to purify the tetanus toxin. One of ordinary skill in the art before the effective filing date of the claimed invention would have been motivated to do this in order to obtain a simple and efficient method for purifying tetanus toxin or variants thereof where Rossetto et al. teach that chromatography on Superose-Zn(II) is shown to be a simple and efficient method for the rapid isolation of tetanus toxin. In view of the reference teachings, it would have been obvious to use all of the method steps, components, pH and elution conditions recited in the claims as routine optimization and/or as desired for purifying tetanus toxin or variants thereof. One of ordinary skill in the art at the time the invention was made would have a reasonable expectation of success because purifying tetanus toxin are known in the art as shown by the above reference teachings especially the teachings of CN116731130A. Hence, the claimed invention as a whole is prima facie obvious.
Conclusion
No claim is allowed.
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/CHRISTIAN L FRONDA/Primary Examiner, Art Unit 1652
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