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 .
Priority
This application is a 371 of PCT/US2020/050056 filed 09/10/2020 which claims benefit of provisional application 63/076,442 filed 09/10/2020 and claims benefit of provisional application 62/898,243 filed 09/10/2019. 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.
Status of the Claims
Claims 8, 11, 12, 14 and 15 are amended.
Claims 1-8, 10-12 and 14-19 are pending (claim set filed 02/27/2026) and are examined on the merits herein.
Withdrawal of Rejections
The response and amendment filed on 02/27/2026 is acknowledged. All of the amendment and arguments have been thoroughly reviewed and considered.
For the purposes of clarity of the record, the reasons for the Examiner's withdrawal and/or maintaining if applicable, of the substantive or essential claim rejections are detailed directly below and/or in the Examiner's response to arguments section.
The previous claims 11, 12, 14 and 15 objection under 35 U.S.C. 112(b) has been withdrawn necessitated by amendment of claim 11, 12, 14 and 15.
The previous claims 11, 12 and 14-19 rejection under 35 U.S.C. 112(b) has been withdrawn necessitated by amendment of claims 11 and 14.
New Rejections
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.
Claims 11, 12 and 14-19 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.
The term “preferentially” in claim 11 is a relative term which renders the claim indefinite. The term “preferentially” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention. It is not clear: (i) whether “preferentially” refers to level of sulfation of chondroitin or to the level of production of sulfated chondroitin relative to other sulfated glycosaminoglycans and (ii) what ratio of chondroitin sulfate to chondroitin or to other sulfated glycosaminoglycans in the product is considered preferential. The scope and boundaries of claim 11 are not certain making claim 11 indefinite.
Claims 12 and 14-19, dependent on claim 11, do not resolve the issue mentioned above and are rejected.
Maintained/Modified Rejections
Claim Rejections - 35 USC § 103
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claims 1-8, 10-12 and 14-19 are rejected under 35 U.S.C. 103 as being unpatentable over He (He et al. Appl. Microbiol. Biotechnol., 2017, 101, 6919-6928 on record in IDS), Trilli (US 20120010399 A1), Longo (Longo et al. Chemico-Biological Interactions, 2016, 259, 39-47), DeAngelis (DeAngelis Glycobiology, 2002, 12, 9R-16R on record in IDS), Deng (US 20040091976 A1) and Glass (US 20200332088 A1 on record in IDS), as evidenced by Hiraoka (Hiraoka et al. J. Biol. Chem., 2000, 275, 20188-20196).
Regarding claims 1 and 11, He teaches expression of human chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris which is exogenous for those microorganisms (Abstract). He teaches that the mammalian cell expression system is not feasible for large scale industrial manufacturing indicating the advantage of using bacterial expression system for making sulfated GAG (p. 6920, right column, 2nd paragraph). The E. coli cells expressing human chondroitin-4-O-sulfotransferase (C4ST-1) were harvested by centrifugation (p. 6921, right column, last paragraph), lysed with sonication, C4ST-1 was purified from the cell lysate (p. 6922, left column, 1st paragraph) and tested for enzymatic activity on the substrate, unsulfated chondroitin, in the presence of sulfo donor, PAPS (p. 6923, left column 3rd paragraph). C4ST-1 appeared to be functional and acting on microbially prepared unsulfated chondroitin producing chondroitin sulfate (Abstract, p. 6924, left column). Although He did not show sulfation of chondroitin in the bacterial cell, it would have been obvious to one of ordinary skill in the art that sulfation of chondroitin is an inherited property of recombinant C4ST-1 and that C4ST-1 will perform the same reaction in the bacterial cell in the presence of its substrates, chondroitin and PAPS. One would have been motivated to assume so with reasonably expected success since C4ST-1 was shown by He to be expressed in bacterial cell and to catalyze in vitro sulfation of chondroitin produced in bacterial cells.
He does not teach reduced expression of kfoE, cysH and proteins forming ATP-binding cassette transporters and does not teach extracting product and isolating sulfated glycosaminoglycans from the product.
Trilli teaches a method of production of chondroitin by recombinant microorganism (Abstract). The method includes culturing of the recombinant microorganism and recovering chondroitin from the broth culture (paragraph 0017) by purification (paragraph 0106). Trilli mentions that chondroitin is defined as linear glycosaminoglycan (paragraph 0003) and chondroitin sulfate is a sulfated glycosaminoglycan (paragraph 0004). Trilli discloses the modification of the microorganism producing glycosaminoglycan, chondroitin, by inactivation or deletion of the gene of endogenous fructosyltransferase: “…subjecting a gene originally present therein, which encodes a protein responsible for the addition of fructose residues to the linear chondroitin back-bone, to inactivation, said inactivation including deletion or substitution entirely or part of said gene or disruption by insertion of an additional nucleotide sequence.” (paragraph 0016). Trilli discloses that the gene is kfoE gene: “…the inactivated gene is the kfoE gene…” (paragraph 0030) and the preferred microorganism is E. coli serotype O5:K4:H4, containing kfoE gene (paragraph 0023).
Longo teaches that sulfotransferases use 3’-phosphoadenosine-5’- phosphosulfate (PAPS) as a sulfate donor (p. 39, left and right columns). Longo discloses the presence in E. coli of another enzyme using PAPS as a substrate, i.e. 3’- phosphoadenosine-5’-phosphosulfite reductase (PAPS reductase) encoded by cysH gene: “In E. coli, PAPS is converted into sulphite (SO32-) and adenosine 3’,5’- bisphosphate (PAP) by the activity of the cysH-encoded PAPS reductase enzyme.” (p. 40, left column, 1st paragraph). Longo teaches that inactivation of cysH gene results in the increase of PAPS concentration (Abstract).
DeAngelis teaches that transport of glycosaminoglycans (GAG) synthesized in E. coli includes multiple distinct polypeptide species from which KpsM and KpsT form the ATP-binding cassette transporter and KpsD and KpsE help to transport GAG across the periplasmic space (p. 14R, left column 2nd paragraph). DeAngelis discloses that mutation of certain Kps results in GAG accumulation in cytoplasm or periplasm (p. 14R, left column, 2nd paragraph).
Deng teaches production glucosamine and N-acetylglucosamine by fermentation in genetically modified microorganism (Abstract). Deng mentions different microorganisms including E. coli (paragraph 0025). Deng discloses collecting and recovering of the end product which can be intracellular or extracellular from the microorganism (paragraph 0228). For intracellular product collecting and recovering is described as removing microorganism from the bioreactor (harvesting microorganism), lysing microorganism by a method that does not degrade the product, such as sonication, osmotic disruption, French press or homogenization, and recovering the end product (e.g. N-acetylglucosamine) by a conventional method such as chromatography, extraction, or membrane separation (paragraphs 0230, 0231, 0234).
Glass teaches methods of making and using substantially pure glycosaminoglycan (GAG) compositions derived from genetically modified cell lines (Abstract). The cell lines are modified to produce specific GAG, e.g. chondroitin sulfate, while being free of other GAGs, such as heparan sulfate (paragraph 0004). Glass teaches purification of GAGs such as heparan sulfate or chondroitin sulfate from cultured and lysed cells and describes different purification techniques including precipitation, filtration, extraction and chromatography for isolation of sulfated GAGs (paragraphs 0075-0077). Glass provides example of extraction of heparin sulfate from the cells or media “using chilled guanidine HCl/Zwittergent extraction buffer with 10 mM EDTA, protease inhibitors (10 mM NEM, 1 mM PMSF, 1 μg/ml pepstatin A, and 0.5 μg/ml leupeptin)” followed by centrifugation and chromatography purification (paragraph 0078).
First, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to add reduction of expression of kfoE gene from Trilli teaching to method of producing sulfated GAG including chondroitin sulfate by bacterial cells expressing sulfotransferase based on He teaching. One would have been motivated to make this combination to increase production of chondroitin sulfate by reducing the alternate reaction of fructosylation of GAG/chondroitin. A skilled artisan would have reasonably expected success in this combination since He described expression of functional sulfotransferase in bacterial cell that can produce sulfated GAG and Trilli disclosed modification to prevent chondroitin from being used in another reaction (fructosylation).
Second, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow Longo guidance on the reduction in activity of cysH gene to increase PAPS concentration for production of sulfated GAG/chondroitin sulfate in E. coli based on teachings of He and Trilli. One would have been motivated to make this modification to provide substrate for sulfotransferase, i.e. PAPS, necessary for production of chondroitin sulfate since otherwise PAPS will be utilized by PAPS reductase. A skilled artisan would have reasonably expected success in this combination since He described expression of functional sulfotransferase in bacterial cell that can produce sulfated GAG and Longo provided method to increase concentration of sulfotransferase substrate and hence increase sulfotransferase activity for efficient production of sulfated GAGs.
Third, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to combine teachings of DeAngelis with teachings of He, Trilli and Longo and add reduction of expression of KpsT, KpsM, KpsD or KpsE proteins involved in GAG transport to method of production of sulfated GAG based on combination of He, Trilli and Longo teachings. One would have been motivated to make this modification to prevent export of GAG from the E. coli cells and hence increase presence of the substrate for production of sulfated GAGs within bacteria since DeAngelis teaches increased accumulation of GAG in the cells caused by modification of transporter proteins. A skilled artisan would have reasonably expected success in this combination since He described expression of functional sulfotransferase in bacterial cell that can produce sulfated GAG and Longo, Trilli and DeAngelis provided methods to increase concentration of sulfotransferase substrates (PAPS and GAG) and hence increase production of sulfated glycosaminoglycans.
Last, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow method of isolation of the intracellular end product N-acetylglucosamine by lysis of E. coli cells and recovery of the product by various methods including extraction as described by Deng and method of isolation of sulfated GAGs described by Glass and apply these methods during production of sulfated GAGs based on combination of He, Trilli, Longo and DeAngelis teachings. One would have been motivated to do that since reduction in the expression of ATP-binding cassette transporters based on DeAngelis teaching leads to accumulation of GAGs in the E. coli cell, introduction of sulfotransferase based on He teaching results in the increased amount of sulfated GAGs inside the E. coli cell and Deng describes different methods of E. coli lysis and end product extraction and Glass provides specific method of isolating sulfated glycosaminoglycans which can be applied to the extract from bacterial cells. A skilled artisan would have reasonably expected success in that since He, Trilli, Longo and DeAngelis provided genetic modifications of the host E. coli cell to increase production of sulfated GAGs and Deng and Glass teach methods of isolation of intracellular products. Thus, combination of teachings of He, Trilli, Longo, DeAngelis, Deng and Glass renders claims 1 and 11 obvious.
Regarding claims 2-4, 18 and 19 Longo teaches expression of cysH encoded PAPS reductase with induction by 0.5 mM IPTG when the cell culture media OD600 reaches 0.6 and at 18° C: “… until OD600 reached 0.6, and proteins' expression was induced with 0.5 mM isopropyl b-D-thiogalactopyranoside (IPTG). … After IPTG addition, bacteria were grown overnight at 18° C under shaking conditions” (p. 40, right column, last paragraph). Longo describes the same OD600 as in claims 2, 3 and 18 and inducer concentration as in claims 3 and 18 and expression temperature in Longo teaching is close to temperature limitations in claims 2, 4 and 19. It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention that the OD600 for the start of protein expression induction and the temperature of the protein expression can be optimized. One would have been motivated to follow Longo guidance and optimize those parameters to increase the level of production of recombinant protein, i.e. sulfotransferase. A skilled artisan would have reasonably expected success in this optimization because selection of OD600 for start of induction and protein expression temperature is routine and conventional. Thus, combination of teachings of He, Trilli, Longo, DeAngelis, Deng and Glass renders claims 2-4, 18 and 19 obvious.
Regarding claims 5 and 16, He teaches expression of chondroitin 4-O-sulfotransferase 1 (Abstract). Thus, He, Trilli, Longo, DeAngelis, Deng and Glass teachings render claims 5 and 16 obvious.
Regarding claims 6 and 17, He discloses that chondroitin-4- sulfotransferase is encoded by Homo sapiens gene C4ST-1 (Abstract). The instant with SEQ ID NO:1 encoding sulfotransferase nucleotide sequence has 100% identity to the nucleotide sequence of C4ST-1 as evidenced by Hiraoka who identified and cloned genes C4ST-1 and C4ST-2 for human chondroitin-4-sulfotransferases 1 and 2, respectively (Hiraoka, Abstract). The GenBank accession number for the C4ST-1 is AF239820 (Hiraoka, p. 20188, left column, 3rd paragraph). Thus, He, Trilli, Longo, DeAngelis, Deng and Glass teachings as evidenced by Hiraoka render claims 6 and 17 obvious.
Regarding claims 7 and 14, Trilli teaches inactivation or deletion of kfoE gene (paragraph 0016) as described for claim 1 above.
Regarding claims 8 and 15, Trilli teaches modified E. coli K4 strain: “the recombinant derivative of Escherichia coli O5:K4:H4 strain U1-41 (from now on referred to as E. coli K4) is obtained by inactivation of the kfoE gene…” (paragraph 0036).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to combine follow Trilli teaching and use modified host E. coli K4 strain with deleted kfoE gene for the method of production of chondroitin sulfate based on He, Longo, DeAngelis, Deng and Glass teachings. One would have been motivated to make this combination with reasonably expected success since Trilli used that strain modification to increase production of chondroitin by reducing the alternate reaction of fructosylation of chondroitin. Thus, He, Trilli, Longo, DeAngelis, Deng and Glass teachings render claims 7, 8, 14 and 15 obvious.
Regarding claim 10, DeAngelis teaches the ATP-binding cassette transporter proteins KpsT, KpsM, KpsD and KpsE as described for claim 1 above. DeAngelis discloses that mutation of certain Kps results in GAG accumulation in cytoplasm or periplasm (p. 14R, left column 2nd paragraph).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow DeAngelis teaching and add reduction of expression of KpsT, KpsM, KpsD or KpsE proteins involved in GAG transport to method of production of sulfated GAG based on combination of He, Trilli, Longo, Deng and Glass teachings. One would have been motivated to make this modification to prevent export of GAG from the E. coli cells and hence increase presence of the substrate for production of sulfated GAGs within bacteria since DeAngelis teaches increased accumulation of GAG in the cells caused by modification of transporter proteins. A skilled artisan would have reasonably expected success in this combination since He provided method for expression of sulfotransferase in bacteria which converted bacterially produced chondroitin into chondroitin sulfate, Trilli described method to prevent alternate reaction of chondroitin fructosylation and Longo and DeAngelis provided methods to increase concentration of sulfotransferase substrates (PAPS and GAG, respectively) and hence increase production of sulfated glycosaminoglycans. Thus, He, Trilli, Longo, DeAngelis, Deng and Glass teachings render claim 10 obvious.
Regarding claim 12, Longo teaches E. coli MG1655 strain for deletion of cysH gene (p. 40, right column, 3rd paragraph).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow Longo teaching and use E. coli MG1655 strain as a host cell for production of sulfated chondroitin based on He, Trilli, DeAngelis, Deng and Glass teachings. One would have been motivated to do that because Longo teaches MG1655 strain for deletion of cysH gene that would increase substrate for sulfotransferase, i.e. PAPS, necessary for production of chondroitin sulfate since otherwise PAPS will be utilized by PAPS reductase. A skilled artisan would have reasonably expected success in this combination since He, Trilli and DeAngelis and Longo describe genetic modifications of the host E. coli cell to increase production of sulfated GAGs. Thus, He, Trilli, Longo, DeAngelis, Deng and Glass teachings render claim 12 obvious.
Response to Arguments
Applicant's arguments filed 02/27/2026 have been fully considered but they are not persuasive.
Applicant argues (addressing p. 6-7 of the Remarks, section 1) that He does not disclose or suggest intracellular production of chondroitin sulfate and He showed that most of produced C4ST-1 was trapped in insoluble form in inclusion bodies and that does not provide motivation to produce sulfated chondroitin in vivo in cell. These arguments are not persuasive because:
He teaches that human sulfotransferase expressed in bacterial cells is fully functional (Abstract) indicating that in the presence of its substrates the recombinant sulfotransferase will inherently possess ability to produce sulfated glycosaminoglycans in bacterial cell. Although He mentions that much of the expressed sulfotransferase was insoluble presumably due to misfolding (p. 6923, right column, last paragraph), He purified sulfotransferase from the soluble fraction and demonstrated its activity of sulfation of chondroitin (p. 6924, left column) confirming presence of sulfotransferase in a soluble form. It is known that expression of eukaryotic proteins in E. coli leads to a pool of partially folded, unfolded or misfolded insoluble proteins since eukaryotic proteins require longer times and/or assistance of chaperones to fold in their native state (p. 5.24.1, right column, last paragraph, p. 5.24.3., left column, 1st paragraph) as evidenced by Francis (Francis and Rage Curr. Prot. Prot. Sci., 2010, 5.24.1-5.24.29). Francis teaches various approaches to increase soluble fractions of expressed proteins one of which is reducing the rates of transcription/translation to allow the newly synthesized protein to fold before it aggregates and forms inclusion bodies. That can be achieved by lowering the temperature of expression (p. 5.24.18, left column, 3rd and 4th paragraphs). He expresses sulfotransferase at 22°C (p. 6921, right column, last paragraph), while Longo is using temperature of 18°C for protein expression as applied for claims 2-4, 18 and 19 in the rejection (p. 40, right column, last paragraph). It would have been obvious to one of ordinary skill in the art that conditions of protein expression in E. coli can be optimized to increase the level of soluble protein produced and that the lower expression temperature will increase the amount of expressed sulfotransferase in soluble form.
Applicant argues (addressing p. 6-7 of the Remarks, section 2) that Trilli discloses method of producing chondroitin by the microorganism and transporting it into the supernatant for separation and purification and combination of He and Trilli teaches away from method of claim 1 and directs one of ordinary skill in the art towards producing of chondroitin sulfate by production and transport of chondroitin according to Trilli and in vitro enzymatic reaction according to He. Applicant adds that “reducing the expression of ATP cassette transporters according to DeAngelis would inhibit the transport of chondroitin required by the method of Trilli, reducing the efficiency of the production of chondroitin, and thus the efficiency of the entire process of producing chondroitin sulfate”. These arguments are not persuasive because:
Trilli teaches increased production of chondroitin by reducing expression of kfoE gene leading to reduction of the alternate reaction of fructosylation of chondroitin (paragraph 016) that provides motivation to combine teaching of He and Trilli to obtain cells producing higher amount of chondroitin due to genetic modification taught by Trilli as a substrate for sulfotransferase expressed based on He teaching resulting in the cell producing chondroitin sulfate. In this case prevention of transport of chondroitin from the cells by reduction of expression of ATP transporters as taught by DeAngelis (p. 14R, left column, 2nd paragraph), will not reduce but on the contrary would increase production of chondroitin sulfate in the cells.
Applicant argues (addressing p. 8 of the Remarks, section 3) that: “Longo describes how sulfation of global regulatory proteins requires PAPS, and that increasing PAPS can have multiple effects on "global cell processes" in E. coli, including "enhancing surface adhesion and cell aggregation," but makes no reference to GAG sulfation.”. These arguments are not persuasive because:
First, although Longo discloses that high concentrations of PAPS can promote production of extracellular structures in E. coli, Longo mentions that: “PAPS promotes cell adhesion and aggregation in sulfate-containing media, while an excess of exogenous cysteine counteracts its effects” (p. right column, 3rd paragraph) indicating that effect of PAPS on production of extracellular structures depends on conditions of E. coli growth medium. Second, although Longo does not describe GAG sulfation, Longo mentions that sulfotransferases use PAPS as a substrate (p. 39). Another enzyme using PAPS as a substrate is PAPS reductase encoded by cysH gene (p. 40, left column, 1st paragraph) and according to Longo deletion of cysH results in PAPS accumulation (Abstract). That provides motivation to add that genetic modification to increase PAPS as a substrate for sulfotransferase and hence increase amount of produced chondroitin sulfate. Lastly, one of ordinary skill in the art would expect that in conditions of expressed sulfotransferase catalyzing sulfation of chondroitin and hence utilizing PAPS, PAPS will not accumulate to the extent to cause significant alterations of cellular structure.
Applicant argues (addressing p. 8 of the Remarks, section 4) that: “Deng discloses the isolation of small molecules from E. coli by fermentation processes involving cell lysis. And Glass discloses methods of isolating GAGs, including sulfated GAGs from eukaryotic cells (not bacteria).” These arguments are not persuasive because:
Deng teaches method of collecting intracellular product from microorganisms including E. coli by lysing cells by a method that does not degrade the product, separating cells and recovering of the product by a conventional method such as chromatography, extraction, or membrane separation (paragraphs 0230, 0231, 0234). That product collection method is general and is applicable not only to small molecules but to sulfated GAGs as well. Although Glass teaches isolation of GAGs from eukaryotic cells, prior art of Glass provides specific purification techniques related to chondroitin sulfate isolation providing motivation to apply it to purification of sulfated GAGs from the lysate obtained as described by Deng.
Applicant argues (addressing p. 8-9 of the Remarks, section 5) that “no combination of the cited prior art references discloses or suggests producing the appropriate sulfotransferase in vivo in the presence of substrates” and that “a PHOSITA would be led by the teachings of the recited references not to the methods of claims 1 and 11, but rather to the simpler method of combining He with Trilli to produce sulfated GAG in vitro according to Trilli, using sulfotransferases according to He. Moreover, given the complex effects of increased PAPS concentrations on "global cell processes," as described by Longo, and the trapping of a large portion of the sulfotransferase enzyme in inclusion bodies as disclosed by He, the ability of the method of claims 1 and 11 to effectively produce GAGs would not be an obvious and predictable result.” These arguments and are not persuasive and were addressed as described above for section 1-3.
Applicant argues (addressing p. 9-11 of the Remarks, sections 6 and 7) that the Office Action used hindsight to select prior art to develop a prima facie case of obviousness and that given the multiple potential complications (effect of increased PAPS concentration on “global cell processes”, trapping of sulfotransferase in inclusion bodies and reduced transport of GAG out of cells can lead to excess of GAG inhibiting intracellular processes prior to effective production of sulfated GAGs), and the availability of an effective process for producing sulfated GAGs by simply combining the in vivo production and transport of GAGs out of the bacterial cell with the in vitro sulfation with GAG-specific sulfotransferase according to He, Applicant respectfully argues that a PHOSITA would not a) be motivated to try the claimed methods, and b) have any reasonable expectation of success that the claimed methods would succeed. These arguments are not persuasive because:
In response to applicant's argument that the examiner's conclusion of obviousness is based upon improper hindsight reasoning, it must be recognized that any judgment on obviousness is in a sense necessarily a reconstruction based upon hindsight reasoning. But so long as it takes into account only knowledge which was within the level of ordinary skill at the time the claimed invention was made, and does not include knowledge gleaned only from the applicant's disclosure, such a reconstruction is proper. See In re McLaughlin, 443 F.2d 1392, 170 USPQ 209 (CCPA 1971). In instant case, one of ordinary skill in the art would be motivated with reasonable expectation of success to combine expression of fully functional sulfotransferase in bacterial cells as taught by He (Abstract) with increase in the amount of chondroitin in the cell by reducing the alternate reaction of chondroitin fructosylation (by kfoE reduction or deletion) as described by Trilli (paragraph 016) expecting that combination to lead to production of sulfated glycosaminoglycans in bacterial cell and further improve the production method by increasing the amount of substrates for sulfotransferase, i.e. PAPS, by reducing expression of cysH as described by Longo (Abstract) and chondroitin by preventing its transport out of cell by reducing expression of ATP transporters based on DeAngelis teaching (p. 14R, left column, 2nd paragraph). One would be motivated to expect production of sulfated GAG in the modified bacterial cells since combination of prior art provides expression of sulfotransferase and increase of concentration of its substrates, chondroitin and PAPS inside the cells. One would be motivated with reasonable expectation of success to isolate produced sulfated GAG by applying methods of bacterial cell harvesting, lysis and product extraction taught by Deng for collection of intracellular products (paragraphs 0230, 0231, 0234) and add specific purification techniques for sulfated glycosaminoglycan on the final purification steps as described by Glass (paragraph 0077).
Therefore, the 35 U.S.C. 103 rejection is maintained.
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 nonprovisional extension fee (37 CFR 1.17(a)) 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.
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/L.G.K./Examiner, Art Unit 1653
/SHARMILA G LANDAU/Supervisory Patent Examiner, Art Unit 1653
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