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 .
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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 6/18/25 has been entered.
Election/Restrictions – Retained for the Record
Applicant’s election without traverse of the species of a purified metallothionein protein having an amino acid identified by SEQ ID NO:6, in the reply filed on 3/11/24 is acknowledged.
Claim 2 is withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected species, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 3/11/24. This is consistent with applicant’s specific request, page 2 of 3/11/24 Remarks.
The Examiner notes that on further consideration and analysis that the sequence of SEQ ID NO:6 excluding the C-terminus histidine tag is a species of the genus of the claim 2 formula.
Claim Status
Claims 1 and 3-17 are pending.
Claim 2 is cancelled.
Claims 1, 3-17 are under examination.
Claims 1, 3-17 are rejected.
Priority
The instant application, filed 01/12/2021 is a Continuation in Part of 14937142 , filed 11/10/2015, now abandoned, claims foreign priority to 103139135, filed 11/11/2014.
Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged. Applicant has not complied with one or more conditions for receiving the benefit of an earlier filing date under 35 U.S.C 120 as follows:
The later-filed application must be an application for a patent for an invention which is also disclosed in the prior application (the parent or original nonprovisional application or provisional application). The disclosure of the invention in the parent application and in the later-filed application must be sufficient to comply with the requirements of 35 U.S.C. 112(a) or the first paragraph of pre-AIA 35 U.S.C. 112, except for the best mode requirement. See Transco Products, Inc. v. Performance Contracting, Inc., 38 F.3d 551, 32 USPQ2d 1077 (Fed. Cir. 1994).
The disclosure of the prior-filed application, Application No. 14937142, fails to provide adequate support or enablement in the manner provided by 35 U.S.C. 112(a) or pre-AIA 35 U.S.C. 112, first paragraph for one or more claims of this application. SEQ ID NO:6, explicitly recited in instant claim 1, was not taught in the prior-filed application, Application No. 14937142, nor, from the examiner’s understanding, in the foreign priority document from which Application No. 14937142 claims priority. The first recitation of SEQ ID NO:6 is in the instant application, which is identified as a Continuation in Part. This sequence, as well as the instant claim 1 step of purifying the metallothionein protein by utilizing cleavage of histidine tag and metal affinity chromatography, are found in instant claim 1, but not in prior applications.
The earliest priority for what is instantly claimed in claim 1 and all claims depending from it is 01/12/2021.
Information Disclosure Statement
The Examiner has considered the reference(s) provided in the 6/18/25 Information Disclosure Statement, and provides a signed and dated copy of such herewith.
Claim Objections
Response to Arguments
Applicant’s response, see page 5, filed 6/18/25, and claim amendments, with respect to the objections to claim(s) 1 and 3 have been fully considered and are persuasive. Therefore, the objection has been withdrawn.
Claim Interpretation
The claim limitations are given their broadest reasonable interpretation (BRI) consistent with the specification, MPEP 2111, and under the BRI, words of the claim must be given their plain meaning, unless such meaning is inconsistent with the specification, MPEP 2111.01.
The transitional term “comprising” is inclusive or open-ended and does not exclude additional, unrecited elements. See MPEP 2111.03.
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 (i.e., changing from AIA to pre-AIA ) 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, 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.
Response to Arguments
Applicant's arguments filed 6/18/25 have been fully considered but they are not persuasive.
Applicant, pages 5-6, argues that the previously applied obviousness rejection is flawed because of the reasoning previously applied with regard to the additional leucine in GenBank AAP36553, stating in part, “the Office does not provide any scientific or technical reasoning why a skilled person would be motivated to remove the leucine residue of GenBank AAP36553.1 in order to carry out the claimed methods. Without such reasoning, a skilled person would not have had an expectation of success. Furthermore, the argument that it would have been obvious to remove the leucine residue appears to be based on hindsight bias.” Without responding to the bases or motivations in the previous rejection, the examiner currently appreciates that given the use of comprises, in claim 1’s “wherein the purified metallothionein protein comprises the amino acid sequence of SEQ ID NO:6,” the presence of any additional amino acids to the N- or C-terminus of the amino acid sequence of SEQ ID NO:6 – these being additional, unrecited elements, is/are not excluded. This simplifies the rejection as modified below.
Also, and related to the arguments and the Declaration and its Appendix I, as set forth in Melcher, there are taught multiple ways to produce a protein such as a metallothionine protein, see Melcher’s para 83, copied below, which includes protein synthesis (so including organic syntheses that do not require use of bacteria, etc., for recombinant protein production). Also, the assertion that “human MTA2A, but not human MT1A, could be produced was completely unexpected” on page 6 refers to a single experiment using Pichia pastoris, and is not considered dispositive of the broad range of possible recombinant approaches with regard to species/cell types, vectors and other factors that influence recombinant protein production. Also, the production by a recombinant microorganism cell is not claimed, so such ‘unexpected’ assertion does not have a direct association to what is being claimed.
With regard to the assertions with respect to new claim 17 that “none of the cited references teach or suggest the additional step of determining the concentration of the purified metallothionein protein by ultraviolet (UV) absorbance at 280 nm. Thus, new claim 17 is not prima facie obvious based on the combination of cited references,” a newly identified reference is combined with the previously applied references to reject claim 17 below.
Applicant argues on page 7 refers back to the Response to Non-Final Office Action submitted on July 26, 2024, regarding features/advantages of inclusion of a His tag, that Response including the “claimed invention addresses the problem of protein quantification during purification by incorporating a poly-histidine tag into the metallothionein (MT) sequence,” and asserting that “This is the first known instance of utilizing a poly-histidine tag for MT, providing a novel method to detect the protein through absorbance measurement at 280 nm. Natural MT sequences lack aromatic amino acids, such as Histidine, Phenylalanine, Tryptophan, and Tyrosine, which are critical for absorbance at this wavelength (280 nm). By adding the poly-histidine tag, the invention not only facilitates protein purification but also introduces a new method for quantifying protein concentration, which was previously unachievable with MT due to the absence of these amino acids. This approach is both innovative and non-obvious in light of prior art.”
As previously stated, the prior art basis for using a histidine tag fused to the MT protein is set forth below, and is silent on the quantification of protein based on the aromatic histidines of the histidine tag. However, the Office’s reasons for combining the prior art, to obtain an improved separation and purification, are sufficient to make and sustain the rejection. That the Office sets forth a different reason for combining prior art teachings to reject a claim than applicant now asserts does not negate the basis for the rejection.
Claim(s) 1, 3-12 and 14-16 are rejected under 35 U.S.C. 103 as being unpatentable over US 20160130325, published 5/12/16, inventors Melcher et al. (Melcher, parent of instant CIP), in view of Honda et al, “One-step purification of metallothionein extracted from two different sources,” Journal of Chromatography B, 820 (2005) 205–210 (Honda), Abdullah and Chase, BIOTECHNOLOGY AND BIOENGINEERING, VOL. 92, NO. 4, NOVEMBER 20, 2005, 501-513 (Abdullah), and GenBank AAP36553.1 (2003), all such references previously applied and prior art references provided, as evidenced by Metallothionein Pfam downloads, two screen shots combined to one page, downloaded 10/15/25 (“Pfam”).
Claim 1 is directed to a method for producing a hypo-metallated redox-active metallothionein protein, the hypo-metallated redox-active metallothionein protein having 20 cysteine sulfhydryl groups and 7 binding pockets, 2 to 16 of the 20 cysteine sulfhydryl groups being free and reduced, and 1 to 6 of the 7 binding pockets being occupied by metal ions, comprising the following steps:
producing a metallothionein protein;
purifying the metallothionein protein by utilizing a histidine tag and metal affinity chromatography;
de-metallating the metallothionein protein;
chemically reducing the de-metallated metallothionein protein; and
partially metallating the reduced de-metallated metallothionein protein with 4 molar equivalents of zinc ions,
wherein the purified metallothionein protein comprises the amino acid sequence of SEQ ID NO: 6.
The elected sequence is SEQ ID NO:6, which includes a six histidine “his tag” at its C-terminus.
Melcher teaches an invention that relates to method for producing hypo-metallated redox-active metallothionein (MT) proteins, pharmaceutical compositions containing the proteins, and uses the pharmaceutical compositions for treatment of conditions originating from elevated intracellular oxidative stress and/or dis-balanced intracellular redox-potential and/or redox-potential-dependent imbalance of metal ions, Abstract.
In some embodiments Melcher’s method steps are as follows:
[0053] The first aspect of the present invention relates to a method for producing a hypo-metallated redox-active metallothionein protein. The hypo-metallated redox-active metallothionein protein has 20 cysteine sulfhydryl groups, thus forming 7 metal ion binding pockets. Two (2) to 16 of the 20 cysteine sulfhydryl groups are free and reduced, and 1 to 6 of the 7 binding pockets are occupied by metal ions. The method comprises the following steps:
[0054] providing a metallothionein protein;
[0055] de-metallating the metallothionein protein;
[0056] chemically reducing all the 20 cysteine sulfhydryl groups of the de-metallated metallothionein protein; and
[0057] partially metallating the reduced de-metallated metallothionein protein by providing 1, 2, 3, 4, 5, or 6 metal ions to the 7 binding pockets.
Melcher teaches that
[0083] The metallothione in apo-proteins of the present invention, in their native form with undefined metallation status and undefined redox-status of their cysteine sulfur residues, may be produced by any conventional method of recombinant DNA technology, protein synthesis, enzymatic cleavage of pre-pro-proteins, or isolation of naturally occurring variants of mammalian metallothionein proteins.
[0084] Thus, in one preferred embodiment of the invention, the metallothionein apo-proteins are produced by established recombinant DNA technology (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, 1989, and references therein). The DNA sequence encoding the respective full-length metallothionein protein isoform may either be synthesized by standard nucleic acid synthesis methods, or may be obtained by established laboratory methods of directly isolating it from appropriate genomic or cDNA libraries, or indirectly by copying and amplifying it from said libraries by standard polymerase chain reaction (PCR) method.
[0085] The DNA sequence, after appropriate preparation and purification, may then be introduced into a recombinant expression vector of choice, using, again, established standard methods. The choice of an appropriate vector may depend on the choice of protein expression system to be used. The expression systems, as established and extensively described in scientific literature well known to the person skilled in the art, include prokaryotic bacterial cells, eukaryotic yeast and fungal cells, as well as cell cultures derived from higher animals such as insects, arthropods or mammals. Introduction of the protein expression vector, equipped with the coding sequence for the chosen MT protein variant, into appropriate host cells, may be facilitated by established protocols.
Therefore Melcher’s step of providing includes preparing per any of the approaches of paras 83-85, and instant claim 1’s first step of producing a metallothionein protein would have been obvious.
Melcher paras 55 and 56 make obvious instant claim 1’s steps 3 and 4.
As to instant claim 1’s step 5, in addition to para 57 above, that statement from the first Example, from para 143, “As representative examples for one preferred embodiment of the invention, the main protein fractions were then partially metallated with 4 molar equivalents of zinc ions (“hypo-MT1a” and “hypo-MT2a”), by adding respective amounts of zinc chloride, based on Bradford protein quantification,” see also para 145, make obvious step 5’s “partially metallating the reduced de-metallated metallothionein protein with 4 molar equivalents of zinc ions,” based on applying a specifically taught amount in the same step.
However, Melcher does not teach step 2 of claim 1, “purifying the metallothionein protein by utilizing of [sic] histidine tag and metal affinity chromatography.”
The level of ordinary skill in the art of purification of peptides and proteins is high.
Honda teaches a “one-step purification of hepatic metallothionein from the Amazon fish Colossoma macropomum injected with cadmium and from the copper-loaded metallothionein from the yeast Saccharomyces cerevisiae, performed by affinity chromatography through metal chelating columns. … hepatic metallothionein extracted from fishes was purified by Ni2+-loaded resin and eluted by a continuous imidazol gradient. Purified metallothioneins were evaluated by SDS–PAGE and characterized by UV spectra of the apo- and Cd2+-loaded protein. This method allowed high purity and yield as well as rapid one-step extraction of both metal-loaded and apoprotein.” Abstract.
Honda teaches that other methods for metallothionein purification generally include several steps, take time, and have a high loss of protein mass, page 205, whereas its method is simpler and only requires a single column, Abstract and page 206, section 2.4.
Based on Honda’s teachings and results, one of ordinary skill in the art would have been motivated to further improve the methods of Melcher by purification that includes metal affinity chromatography. The motivation is to purify metallothioneins with a simpler, faster method that provides greater yield by having less protein losses. There would have been a reasonable expectation of success given the teachings and results of Honda, including for metallothioneins from a fish species.
Honda teaches that affinity chromatography “has been a widespread and powerful technique for purification of mainly recombinant proteins modified with specific sequences, e.g., polyhistidine tails, enzymes, monoclonal antibodies, DNA-binding proteins and so on,” page 209, but does not extensively teach the advantages of utilizing a poly-histidine tag for purification procedures.
Abdullah teaches that poly-histidine tags, “fused to either the N- or C-terminal end of the target protein and hence provides a strong selective affinity for binding to metal ions and allows histidine tagged proteins to be separated using the immobilized metal affinity chromatography (IMAC) strategy. IMAC separation works efficiently under mild operating condition is low in cost, robust, and has high ligand selectivity and capacity, thus making it suitable for large-scale applications,” (citation omitted), page 502.
Thus Abdullah expands on the single statement in Honda regarding “polyhistidine tails”, and teaches that this provides a strong selective affinity for binding to metal ions and allows histidine tagged proteins to be separated using the immobilized metal affinity chromatography. This clearly supports that metal affinity chromatography utilizing histidine tags was commonly practiced and common in the art, so that one of ordinary skill in the art wanting to improve purification and separation of a desired protein would reasonably consider applying his-tag separation and purification. The rationale is to improve the process of Melcher to obtain a desired purified metallothionein by incorporating a Histidine tag, as routinely used in the art to obtain a simpler purification with improved yield. There would have been a reasonable expectation of success given the teachings and results of Abdullah, also considering the teachings of Honda.
None of Melcher, Honda or Abdullah explicitly teach the specific sequence of SEQ ID NO:6’s first 61 amino acids.
However, Melcher clearly teaches producing and purifying and using metallothionein 2a (MT2a), see paras 69-72, 106 with SEQ ID NO:4 referred to as bovine metallothionein 2a protein (MT2a), 143, 145-150.
Melcher’s SEQ ID NO:4 compared with instant SEQ ID NO:6 is as follows:
Query Match 95.5%; Score 364; DB 1; Length 67;
Best Local Similarity 91.8%;
Matches 56; Conservative 3; Mismatches 2; Indels 0; Gaps 0;
Qy 1 MDPNCSCTAGESCTCAGSCKCKDCKCASCKKSCCSCCPVGCAKCAQGCVCKGASDKCSCC 60
||||||| ||:|||||||||||:||| |||||||||||||||||||||:|||||||||||
Db 1 MDPNCSCAAGDSCTCAGSCKCKECKCTSCKKSCCSCCPVGCAKCAQGCICKGASDKCSCC 60
Qy 61 A 61
|
Db 61 A 61
Melcher teaches multiple uses of its processed metallothioneins for treating human subjects having a variety of diseases/conditions, see paras 5, 7-9 (providing background), 92, teaching alternatives of bovine and human MT isoforms, and 120-132, see also claims 10-15, so one of ordinary skill in the art would have considered substituting a similar human MT2a for Melcher’s bovine MT2a at least because a human MT2a would reasonably have greater chance of greater tolerance and effectiveness than a bovine MT2a when administering to a human subject.
Among known human metallothionein 2A sequences is a 62 amino acid sequence, identified as a partial synthetic construct, that is 100% identical with SEQ ID NO:2’s first 61 amino acids (i.e., without the His tag), this provided by Kalnine et al., submitted to GenBank 5/13/2003, having the following sequence:
mdpncscaag dsctcagsck ckeckctsck ksccsccpvg cakcaqgcic kgasdkcscc al (GenBank AAP36553.1, provided).
The only difference between this sequence and instant SEQ ID NO:6 excluding the His tag is that the GenBank sequence comprises a 62nd amino acid, Leucine. However, GenBank AAP36553.1 clearly teaches that the region of 4 to 61, which excludes the leucine at position 62, is the region having the name “Metallothio” that belongs in pfam00131. Pfam00131 comprises a group of sequences of different species having a common core that is identified as having metallothionein function, see evidentiary reference Pfam. At a minimum, this suggests to one of ordinary skill in the art that the C-terminus leucine is not required for metallothionein activity, and a person of ordinary skill in the art would reasonably understand that the leucine at position 62 is not expected to be required for metallothionein function or activity based on this distinction. That is, GenBank AAP36553.1 states that from the sequence between 4 to 61, inclusive of these end amino acids, is that portion of the entire sequence that belongs within the pfam00131 of methallothioneins, and is responsible for metallothionein activity, so the C-terminus leucine would not be required for such metallothionein activity.
One of ordinary skill in the art seeking a human MT2a sequence instead of the bovine MT2a taught by Melcher, to better treat human diseases/conditions, would have identified the human MT2A sequence of GenBank AAP36553.1, and would have considered substitution of this for the bovine sequence of Melcher. Based on the above teachings in GenBank AAP36553.1, and as further evidenced by Pfam, in developing a desired metallothionein for therapeutic use the C-terminal leucine would be understood by one of ordinary skill in the art to not be required for function. The motivation to not include this C-terminus lysine is to provide a human metallothionein-derived sequence that comprises the sequence providing the metallothionein function, without amino acids not required for such function or for other functions (e.g., methionine as the first amino acid in an expressed sequence) and is derived from a human metallothionein so as to afford potentially improved performance when treating human diseases/conditions, including, additionally, with less risk of potential adverse immune response.
As an additional basis, when considering that Melcher SEQ ID NO:4 terminates with SCCA, without a leucine, one of ordinary skill in the art would have found additional support for there being no need for the terminal leucine of GenBank AAP36553.1. That is, this teaching and example of Melcher further supports that the C-terminus leucine of GenBank AAP36553.1 would not be required for metallothionein activity.
There would have been a reasonable expectation of success given the description in GenBank AAP36553.1 of the 4..61 portion being that portion in the metallothio pfam00131 – so indicating desired function, and also the teachings of Melcher including that of SEQ ID NO:4 and as to use of human MT2a. Additionally, it is not inventive to substitute a known human protein for a previously taught bovine protein of the same type and function when considering use of that protein in therapies for humans, and to not include any amino acid of a reported human sequence that is not attributed to the desired function, here the metallothionein function.
Accordingly, claim 1 would have been obvious, as would the elected species SEQ ID NO:6.
Claim 3 would have been obvious based on the teachings in references applied to claim 1, specifically including Melcher’s teachings of fully pre-metallating, see para 59, and Melcher claim 3, and of magnetic force purification including when fully pre-metallated, paras 96 and 97.
Claim 4 would have been obvious based on the additional teaching of using zinc sulfate or zinc chloride, “…in order to allow for full pre-metallation. From this solution, the fully reconstituted Zn.sub.7-MT protein may be recovered using magnetic force,” para 97 of Melcher.
Claim 5 would have been obvious based on the teachings in references applied to claim 1, and the additional Melcher claim 4, having substantially the same language as instant claim 5.
Claim 6 would have been obvious based on the teachings in references applied to claim 1, and specifically Melcher’s para 99 teachings, these including relevant portions of claim 6 limitations.
Claims 7 and 8 would have been obvious based on the teachings in references applied to claim 1, and specifically Melcher’s para 100 teachings combined with Melcher claims 5 and 6.
Claims 9 and 10 would have been obvious based on the teachings in references applied to claim 1, and specifically Melcher’s para 101 teachings combined with Melcher claims 7 and 8.
Claims 11 and 12 would have been obvious based on the teachings in references applied to claim 1, and specifically Melcher’s para 102 teachings.
Claim 14 would have been obvious based on the teachings in references applied to claim 1, and specifically Melcher’s para 82 teachings combined with Melcher claims 11 and 12.
Claim 15 would have been obvious based on the teachings in references applied to claim 1, and specifically Melcher’s paras 128, 148-150 and Fig. 3A and 3B.
Claim 16 would have been obvious based on the teachings in references applied to claim 1, and specifically Melcher’s paras 115, 121-130, and 151-154.
Claim(s) 13 is rejected under 35 U.S.C. 103 as being unpatentable over US 20160130325, published 5/12/16, inventors Melcher et al. (Melcher, parent of instant CIP), in view of Honda et al, “One-step purification of metallothionein extracted from two different sources,” Journal of Chromatography B, 820 (2005) 205–210 (Honda), Abdullah and Chase, BIOTECHNOLOGY AND BIOENGINEERING, VOL. 92, NO. 4, NOVEMBER 20, 2005, 501-513 (Abdullah), and GenBank AAP36553.1 (2003), as evidenced by Metallothionein Pfam downloads, two screen shots combined to one page, downloaded 10/15/25 (“Pfam”), as applied to claim 1 above, and as evidenced by GE Instructions 11-0008-88 AH, 24 pages, 2014 (GE).
The rejection of claim 1 is set forth above.
Claim 13 depends from claim 1 and states “wherein the metal affinity chromatography utilizes a nickel (Ni++) prepacked column.
Honda teaches using a HiTrapTM Chelating HP column that comprises Ni2+, and was previously loaded, page 206 sections 2.2 and 2.4, see also page 207 bottom right column, but does not explicitly state that this was prepacked.
As noted above, the level of ordinary skill in the art of purification of peptides and proteins is high.
GE evidences that “HisTrap FF is a ready to use HiTrap™ column, prepacked with precharged Ni Sepharose™ 6 Fast Flow. This prepacked column is ideal for preparative purification of histidine-tagged recombinant proteins by immobilized metal ion affinity chromatography (IMAC).
HisTrap FF columns provide fast, simple, and easy separations in a convenient format, including an excellent start for scaling up.”
GE evidences that the column used by Honda was prepacked, so claim 13 would have been obvious over the rejection applied to claim 1 above, with the evidentiary reference GE supporting the rejection as to the prepacked limitation.
Claim(s) 17 is rejected under 35 U.S.C. 103 as being unpatentable over US 20160130325, published 5/12/16, inventors Melcher et al. (Melcher, parent of instant CIP), in view of Honda et al, “One-step purification of metallothionein extracted from two different sources,” Journal of Chromatography B, 820 (2005) 205–210 (Honda), Abdullah and Chase, BIOTECHNOLOGY AND BIOENGINEERING, VOL. 92, NO. 4, NOVEMBER 20, 2005, 501-513 (Abdullah), and GenBank AAP36553.1 (2003), as evidenced by Metallopthionein Pfam downloads, two screen shots combined to one page, downloaded 10/15/25 (“Pfam”), as applied to claim 1 above, and further in view of Quantifying protein using absorbance at 280 nm, Experimental Biosciences web page from www.rice.edu, 2015, 3 pages, downloaded from the internet 10/14/25 (Rice).
The rejection of claim 1 is set forth above.
Claim 17 depends from claim 1 and further comprises determining the concentration of the purified metallothionein protein by ultraviolet (UV) absorbance at 280 nm.
None of the references applied to claim 1 explicitly teach this claim 17 step.
As noted above, the level of ordinary skill in the art of purification of peptides and proteins is high.
Rice teaches that “Quantifying protein by directly measuring absorbance is fast and convenient, since no additional reagents or incubations are required. No protein standard need be prepared and the procedure does not consume the protein,” page 1. Rice also teaches specific steps for determining protein content by measuring ultraviolet (UV) absorbance at 280 nm, page 2, Procedure.
It would have been obvious to apply the method of Rice to measure protein content as set forth in claim 17 because this method is fast, convenient, and does not consume the protein. There would have been a reasonable expectation of success because the method as taught by Rice is commonly used for the same or similar purpose as set forth in Rice, “The most common use for this method is to monitor fractions from chromatography columns, or any time a quick estimation is needed and error in protein concentration is not a concern.”
Accordingly, claim 17 would have been obvious and is rejected under this section.
Conclusion
No claim is allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to JOSEPH FISCHER whose telephone number is (571)270-7925. The examiner can normally be reached on Monday to Friday, 9:00 AM to 5:00 PM, however noting that the examiner will not normally be working on Wednesday-Friday and on Monday/Tuesday on alternating weeks, but will promptly answer messages upon his return to work.
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, MELISSA FISHER, can be reached on 571-270-7430. 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.
/JOSEPH FISCHER/Examiner, Art Unit 1658