DETAILED ACTION
Notice of Pre-AIA or AIA Status
1. The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Preliminary Claim Amendment
2. The amendment filed October 27, 2025 was entered. Claims 1-14 were cancelled. Claims 15-29 were newly added. Claims 15-29 are under consideration in this Office Action.
Information Disclosure Statement
3. The information disclosure statement (IDS) submitted on January 17, 2025 was filed. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Claim Objections
4. Claims 19-22, 25-26 and 29 were objected to because of the following informalities:
A) Claims 19-20 recites “…in an amount of25-500mg…”, clarification is required to overcome the objection.
B) Claim 21 recites “… a diarrheogenic Escherichia coli spp. (enterotoxigenic and enterohemorrhagic) antigen…” First, “diarrheogenic” and “enterohemorragic” are misspelled, and should be spelled diarrheagenic and enterohemorrhagic. Second, diarrheagenic E coli includes enterotoxigenic and enterohemorrhagic E coli as primary categories of diarrheagenic E coli.
C) Claims 22 and 25 recites FeCl3, H2O2; however the correct chemical signature is FeCl3, H2O2. Therefore, clarification is required to overcome the objection.
D) Claim 26 was recite (‘CDAP’) and (‘CTEA’) It is unclear what the apostrophe and parentheses are for.
E) Claims 25 and 29 respectively recite “…(DT)is..” and “…(DT)may..” Thus, clarification is required to overcome the objection.
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.
5. Claims 15-20 and 24-26 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.
a) Claims 15-20 and 24 separates the method steps using a period. Where a claim sets forth a plurality of elements or steps, each element or step of the claim should be separated by a line indentation, 37 CFR 1.75(i). There may be plural indentations to further segregate subcombinations or related steps. Therefore, the periods should be removed and only the period at the completion of the claim should remain. See MPEP 608.01(m).
b) Claim 15(a) recites “…improved harvest yield (100-700 mg/L)…” However it is unclear if the claim is drawn to the improved harvest yield actually being between 100-700 mg/L or if an example of the improved yield is between 100-700 mg/L, thus the 100-700 is not a required limitation. Therefore the metes and bounds of the claim are unclear with regard to limitations are or are not included as a claim limitation. See also claim 15(b) with EDTA (4 to 10mM), Sodium acetate (5% to 10%); alcohol precipitation (30% to 60%); reduction of endotoxin (< 100EU of endotoxin per µg of PS), protein (< 1%) and nucleic acid 2%) impurities, higher recovery of capsular polysaccharide suitably in the range of 40% to 65%, with the desired O-acetyl levels (> 2.0 mmol/g polysaccharide), Molecular size distribution (>50% of PS is eluted before a distribution coefficient (KD) of 0.25 is reached). See also claim 24 which has the same issues.
c) Claim 15(a) recites a “high yield harvest” and “improved harvest yield” . However the metes and bounds for determining “high” and “improved” harvest yield is not clear. It is unclear how to determine high or improved yield when there is no comparison recited by the instant claims. Claim 15(b) recites “desired” however it is unclear what the metes and bounds for defining what the “desired” levels are or what levels of undesirable levels are. Thus, clarification is required to overcome the rejection. See also, “significant reduction”, “higher recovery”, and “substantially all” all recited by claim 15.
d) Claims 17, 18, 19, 20 and 25-26 recites alternative limitations which are improperly expressed. Alternative expressions are permitted if they present no uncertainty or ambiguity with respect to the question of scope or clarity of the claims. One acceptable form of alternative expression, which is commonly referred to as a Markush group recites members as being “selected from the group consisting of A, B and C”. Another acceptable form recites “selected from 1, 2, 3, or 4.” Applicant may correct this by amending the claim to recite the appropriate language.
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.
6. Claims 15-29 are rejected under 35 U.S.C. 103 as being unpatentable over Royal (CN102935226 published 2013-02-20; priority 2012-11-16) in view of Peddireddy et al., (EP2952586 published 2015-12-09; priority to 2015-06-01).
The claims are drawn to a method of manufacturing a Salmonella enterica serovar typhi Vi polysaccharide-tetanus toxoid conjugate antigen.
Royal teach a typhoid fever and paratyphoid fever combined vaccine. The typhoid fever and paratyphoid fever combined vaccine comprises typhoid Vi polysaccharide and paratyphoid fever thallus specific polysaccharide O-SP. The invention further discloses a preparation method of the vaccine. The preparation method comprises the steps of preparing the typhoid Vi polysaccharide, preparing the paratyphoid fever thallus specific polysaccharide O-SP, preparing typhoid Vi polysaccharide-carrier protein conjugate, preparing paratyphoid fever thallus specific polysaccharide O-SP-carrier protein conjugate and preparing end products [abstract]. Preferably, above-mentioned typhoid fever thalline capsular polysaccharide and paratyphoid A thalline specific polysaccharide O-SP all form conjugate with carrier protein. Preferably, carrier protein is tetanus toxoid [Summary of Invention].
The preparation method of above-mentioned typhoid fever paratyphoid A combined vaccine, comprises the following steps, 1) refining typhoid Vi polysaccharide: the Salmonella typhimurium strain is after the formaldehyde sterilization, and high speed centrifugation is removed thalline, adds cetyl trimethyl ammonium bromide in supernatant, fully mixes and forms precipitation, and centrifugal collecting precipitate, use CaCl 2Solution dissociates, then makes semifinished product by the ethanol precipitation; Semifinished product is dissolved in 10% saturated acetic acid sodium solution, then uses cold phenol extraction, centrifugal collection supernatant, and dialyse with the 0.1mol/L calcium chloride solution; Adding ethanol to ultimate density is 75～80% (V/V) again; Centrifugal collecting precipitate, centrifugal after washing, abandon supernatant, the polysaccharide of dissolution precipitation, obtain refining typhoid Vi polysaccharide;
2) prepare paratyphoid A thalline specific polysaccharide O-SP: the Salmonella paratyphi A kind is after the formaldehyde sterilization, and high speed centrifugation is collected thalline, the secondary formicester polysaccharide in the rough thalline of hot phenol method; With the refining secondary formicester polysaccharide of the cold ethanol sedimentation method, then secondary formicester polysaccharide is dissolved in 1% acetum with 10mg/ml concentration again, cooling after boiling water bath, the centrifugal precipitation of abandoning, the dialysis supernatant, then with cold ethanol sedimentation method collecting precipitation, through the molecular sieve chromatography purification;
3) prepare typhoid Vi polysaccharide-carrier protein conjugate: the derivation carrier protein; To make with extra care typhoid Vi polysaccharide solution and add carbodiimide, then add the carrier protein of derivation, and maintain pH and carry out association reaction 5.75, then adjust pH to neutral cessation reaction, reactant is dialysed overnight in normal saline, crosses the chromatographic column purification, obtains typhoid Vi polysaccharide-carrier protein conjugate;
4) prepare paratyphoid A thalline specific polysaccharide O-SP-carrier protein conjugate: with adipic dihydrazide and paratyphoid A thalline specific polysaccharide O-SP, carry out coupled reaction, make the O-SP-AH derivant, it is mixed with carrier protein, add carbodiimide to carry out association reaction, remove small-molecule substance through the ultrafilter membrane ultrafiltration, cross the chromatographic column purification, obtain paratyphoid A thalline specific polysaccharide O-SP-carrier protein conjugate;
5) prepare finished product: by typhoid fever thalline capsular polysaccharide-carrier protein conjugate stock solution with add autoclaving after paratyphoid A thalline specific polysaccharide O-SP-carrier protein conjugate stock solution is mixed after injection water and lactose, lyophilizing after aseptic filtration [Summary of Invention]. The invention has the beneficial effects as follows: the present invention is to the immunity simultaneously of typhoid fever and paratyphoid A, and noiseless between two kinds of antigen, all can produce the immune response identical with independent conjugate [Summary of Invention].
Embodiment 3 recites The preparation method of typhoid Vi polysaccharide-carrier protein conjugate: Using the carrier protein of tetanus toxoid (TT) as typhoid Vi polysaccharide. 1. TT derivation: TT is pressed to the protein content dilution for 10mg/ml, add 3.5 milligram 1 at 20～26 ℃ of every milligram of albumen of room temperature, 6-adipic dihydrazide (ADH), mix, adjust pH to 5.75, adding carbodiimide (EDAC) to make the ratio of EDAC and albumen is 0.1-0.8, maintains pH5.75 and stirs 1 hour, with the ultrafiltration of 30k ultrafilter membrane, removes the small molecular weight impurities such as ADH and EDAC. Measure protein content and ADH residue (AH) content, calculate its derivation rate. During derivation albumen, EDAC/TT fixes between 0.1～0.15, and more suitable derivation degree can be arranged.
2. Typhoid Vi polysaccharide is combined with the derivation carrier protein: by 0.1mol/l HCl adjust pH to 5.75 for typhoid Vi polysaccharide, by adding volume after albumen to add the EDAC of 5～10mmol/l, adjust pH to 5.75, react 2 minutes, add isopyknic derivation albumen TT-AH, maintain pH 5.75, react 3 hours or pH while no longer changing, adjust pH to neutral cessation reaction. Reactant is dialysed overnight in normal saline, crosses Sepharose 4FF column purification, collects first peak, removes floating proteins and other small-molecule substance. When polysaccharide and albumen coupling, high concentration and the easy plastic of high derivation degree conjugate, and be difficult for combination when low concentration and low derivation are spent, coupling preferably and suitable GL-PP ratio are only just arranged when suitable concentration. Albumen derivation degree can be avoided plastic at 1.0-3.0%, GL-PP final concentration during at 2-3mg/ml, and GL-PP ratio is preferably arranged again [Embodiment 3]. Therefore Royal teach claims are drawn to a manufacturing a Salmonella enterica serovar typhi Vi polysaccharide-tetanus toxoid conjugate antigen but does not teach the specific culturing techniques.
Peddireddy et al., teach cost-effective manufacturing processes for such vaccines have become a priority. Increasing capsular polysaccharide "fermentation harvest stage yield" by employing novel feed strategies and fermentation medium has been one of the preferable approach to achieve said objective. However all the available fermentation processes previously disclosed fail to improve "fermentation harvest stage yield" of N. meningitidis capsular polysaccharide beyond 1.5 g/L. Also formation of foam during large scale fermentation could i) reduce capsular polysaccharide yield due to loss of cells and culture medium to the foam phase, ii) can be detrimental to cells since when bubbles burst they exert sheer forces iii) result in a loss of sterility if the foam escapes and iv) can lead to over-pressure if a foam-out blocks an exit filter [para 6]. To prevent the formation of foam, mechanical foam breakers, ultrasound or, most often, the addition of chemical antifoaming agents (or "antifoams") are routinely employed [para 7]. These include their effects on foam height with time, their influence on the volumetric oxygen mass transfer coefficient (kLa) of the system, their gas hold-up characteristics and their globule size and distribution in relation to their action upon foams [para 8].
Peddireddy et al., teach gradual addition of a 10%v/v to 20% v/v solution of an antifoam, being an alkoxylated fatty acid ester on a vegetable base added at a final concentration from 0.01% to 0.1% to Neisseria meningitides A cultures from 4th hr of log phase onwards till 11th hr i) increase the OD590 by 5 units, thereby obtaining OD590 of about 22 to 27 in comparison to OD590 of about 15 to 17 observed for fermentation cultures without this antifoam, ii) extends the duration of log phase iii) thereby obtaining significantly improved capsular polysaccharide yield of about 2 g/L to 2.5 g/L at fermentation harvest stage such that the polysaccharide at fermentation harvest stage has minimum load of contaminants like protein and nucleic acid [para 12]. The N. meningitidis A polysaccharide yield at fermenter harvest stage can be at least 2 fold greater than the yield reported in prior art [para 18].
The fermentation medium used in the process of the invention having pH from 7 to 7.1 , comprises dextrose from 8 g/L to 10 g/L, sodium chloride from 5.6 g/L to 6 g/L, dipotassium sulfate from 0.8 g/L to 1 g/L, arginine from 0.2 g/L to 0.4 g/L, serine from 0.4 g/L to 0.6 g/L, cysteine from 0.20 g/L to 0.25 g/L, magnesium chloride (MgCl2) from 0.18 g/L to 0.20 g/L, calcium chloride (CaCl2) from 0.020 g/L to 0.022 g/L, ferrous sulphate (FeSO4) at 0.002 g/L, L-glutamic acid from 4.8 g/L to 5.2 g/L, soya peptone from 8.8 g/L to 9.2 g/L, ammonium chloride (NH4Cl) from 0.44 g/L to 0.46 g/L, di-potassium hydrogen phosphate (K2HPO4) from 3.8 g/L to 4.2 g/L and yeast extract from 5.30 g/L to 5.35 g/L. The fermentation method of the present invention uses a feed solution comprising dextrose from 70 g/L to 75 g/L, mono sodium glutamate from 37 g/L to 37.5 g/L, arginine from 2.8 g/L to 3.2 g/L, serine from 2.8 g/L to 3.2 g/L, magnesium chloride (MgCl2) from 1.8 g/L to 2.2 g/L, calcium chloride (CaCl2) from 0.13 g/L to 0.15 g/L, ferrous sulphate (FeSO4) at 0.02 g/L, soya peptone from 4.8 g/L to 5.2 g/L, cysteine from 1.8 g/L to 2.2 g/L and yeast extract from 8.2 g/L to 8.4 g/L. An important aspect of present invention is that antifoam is gradually added as a 10%v/v to 20% v/v solution at a final concentration from 0.01% to 0.1% to N. meningitides A cultures from 4th hr of log phase till 11th hr thereby resulting in i) an OD590 of about 22 to 27 in comparison to OD590 of about 15 to 17 observed for other antifoams, ii) extended duration of log phase iii) minimum level of protein, nucleic acid contaminants. Preferably said antifoam addition can begin at 4th hr of fermentation at a rate of about 20 ml /hr initially and thereafter at a rate of about 35- 50 ml /hr [para 20]. Peddireddy et al., provides a novel fermentation method for obtaining higher N. meningitidis serogroup A capsular polysaccharide yield of about 2 to 2.5 gm/L at fermentation harvest stage by gradual addition of an antifoam agent along with an optimal feed solution from log phase onwards, characterized in that an antifoam comprising of alkoxylated fatty acid ester on a vegetable base is metabolized as a nutrient by N. meningitidis thereby providing at least 2 fold polysaccharide yield improvement as compared to i) yield observed for fermentation cultures without this antifoam or ii) yield observed for fermentation cultures employing polyalkylene glycol based surfactants as antifoam [para 21].
The fermentation procedure includes seed vial containing 3 ml of N. meningitidis A - strain culture having OD 1/ml was frozen at -70°C. Then vial was thawed and seeded into 40 ml of seed medium which was incubated at 37°C and agitated at 150 to 180 rpm. The 40 ml seed culture with OD > 1.0 ± 0.2 was directly inoculated to 800 ml seed medium and the culture was again incubated at 37°C under agitation 160 -200 rpm, till the OD reached > 1.0 ± 0.2. Seed Culture having OD 1.0 ± 0.2 with volume 800 ml was seeded into the reactor, wherein the fermentation medium volume in the reactor was ~ 12L- 14L. After inoculation of reactor 0 hr OD was maintained at 0.04 to 0.06.pH during the fermentation process was maintained around 7.1.The entire fermentation process was carried out in a 30L Stainless steel bioreactor, wherein fermentation cycle was run in a continuous fed batch mode and total duration of fermentation cycle was around 12-18hrs [para 22]. The fermentation process was terminated once drop in optical density was observed followed by inactivation using 1-2% formaldehyde for about 2hrs to 3hrs at 37°C. Further the temperature was reduced to 10°C and incubated for 30 minutes. The harvest was unloaded and centrifuged at 14,500g for 45-60 minutes the inactivated cell pellet was discarded and the supernatant was subjected to 0.2u sterile filtration followed by 100KD diafiltration (10-15 times) and is further concentrated with WFI [para 23]. Table 1 shows the size of the harvest polysaccharide 900-1400 kDa.
A mixture of 1% deoxycholate, 6% sodium acetate, 2mM EDTA & 40% ethanol was added to the 100KD diafiltered harvest. Then the mixture was kept at 2―8°C for 3-4hrs with stirring. Later mixture was subjected to centrifugation at 10000 rpm for 20min and supernatant was diafiltered against 25 mM Tris with 100KD cassette membrane. Further 3% w/v cetyltrimethylammonium precipitation was carried overnight at 2―8°C with stirring and pellet was collected. Said pellet was dissolved in 96% ethanol with 0.05M NaCl at 2―8°C for 2 hrs on stirring. Then polysaccharide precipitation was carried for 30 minutes and pellet was collected. Said pellet was dissolved in 45% ethanol with 0.4M NaCl for 1hr. The supernatant was collected & filtered through CUNO R32SP carbon filter. Then polysaccharide was precipitated in 96 % ethanol for 1-2 hrs and pellet was collected. Then pellet was dissolved in WFI followed by TFF. Final purified N. meningitidis A polysaccharide was stored at -20° C [para 25]. Peddireddy et al., provide Table 2 shows Polysaccharide recovery, Polysaccharide content, O-Acetyl content, protein impurity, nucleic acid impurity, endotoxin molecular size distribution.
Therefore, it would have been prima facie obvious at the time of applicants invention to modify the method of manufacturing a Salmonella enterica serovar typhi Vi polysaccharide-tetanus toxoid conjugate antigen as taught by Royal and incorporating a optimal feed solution as taught by Peddireddy et al., comprising the advantageous combination to increase the optical density and thereby enhance yield to above 2 g/L at fermentation harvest stage. One of ordinary skill in the art would have had a reasonable expectation of success by incorporating defoaming agents for enhancing yields of recombinant proteins in prokaryotic and eukaryotic systems. Moreover, no more than routine skill would have been required to modify the method as taught by Peddireddy et al., by obtaining significantly improved capsular polysaccharide yield at fermentation harvest stage such that the polysaccharide at fermentation harvest stage has minimum load of contaminants like protein and nucleic acid.
It is noted, that while the references recite specific ranges; neither specifically recite the stated ranges. Regarding the specific ranges recited in the instant claims, MPEP 2144.05 states, "[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955) (Claimed process which was performed at a temperature between 40°C and 80°C and an acid concentration between 25% and 70% was held to be prima facie obvious over a reference process which differed from the claims only in that the reference process was performed at a temperature of 100°C and an acid concentration of 10%.); see also Peterson, 315 F.3d at 1330, 65 USPQ2d at 1382 ("The normal desire of scientists or artisans to improve upon what is already generally known provides the motivation to determine where in a disclosed set of percentage ranges is the optimum combination of percentages."); In re Hoeschele, 406 F.2d 1403, 160 USPQ 809 (CCPA 1969) (Claimed elastomeric polyurethanes which fell within the broad scope of the references were held to be unpatentable thereover because, among other reasons, there was no evidence of the criticality of the claimed ranges of molecular weight or molar proportions.). For more recent cases applying this principle, see Merck & Co. Inc. v. Biocraft Laboratories Inc., 874 F.2d 804, 10 USPQ2d 1843 (Fed. Cir.), cert. denied, 493 U.S. 975 (1989); In re Kulling, 897 F.2d 1147, 14 USPQ2d 1056 (Fed. Cir. 1990); and In re Geisler, 116 F.3d 1465, 43 USPQ2d 1362 (Fed. Cir. 1997)."
Additionally, KSR International Co. v. Teleflex Inc., 127 S. Ct. 1727, 1741 (2007), discloses combining prior art elements according to known methods to yield predictable results, thus the combination is obvious unless its application is beyond that person's skill. KSR International Co. v. Teleflex Inc., 127 S. Ct. 1727, 1741 (2007) also discloses that "The combination of familiar element according to known methods is likely to be obvious when it does no more than yield predictable results". It is well known to take a method of manufacturing where there is no change in the respective function of the reagents; thus the combination would have yielded a reasonable expectation or success along with predictable results to one of ordinary skill in the art at the time of the invention. Accordingly, it would have been obvious to a person of ordinary skill in the art to combine prior art elements according to known methods that is ready for improvement to yield predictable results. The claimed invention is prima facie obvious in view of the teachings of the prior art, absent any convincing evidence to the contrary.
Pertinent Art
7. The prior art made of record and not relied upon is considered pertinent to applicant’s disclosure. Biemans et al., (WO 2008081022 priority to 1008-01-02) teach the conjugation of saccharides (in particular the Vi capsular saccharide from Salmonella typhi) and proteins using carbodiimide condensation. It also relates to immunogenic compositions that may be made comprising the saccharide-protein conjugates of the invention. Levine et al., (US 20130129776 ) teach multivalent Salmonella enterica serovar conjugate vaccines comprising conjugates of S. Typhimurium, S. Enteritidis, S. Choleraesuis, S. Typhi, S. Paratyphi A and optionally S. Paratyphi B, wherein the conjugates comprise a hapten antigen and a carrier antigen, wherein at least one of the hapten antigens or carrier antigens is characteristic of the Salmonella enterica serovar. Costantino et al., (WO 2009081276 published 2009-07-02) teach the optimization of culture conditions to improve the production of bacterial capsular polysaccharides from Streptococcus strains in fed batch culture and to novel purification methods suitable for production scale purification of bacterial capsular polysaccharides from Streptococcus strains resulting in higher levels of purity than previously obtained for production scale. See also, AU201302943.
Conclusion
8. No claims allowed.
9. Any inquiry concerning this communication or earlier communications from the examiner should be directed to JA-NA A HINES whose telephone number is (571)272-0859. The examiner can normally be reached Monday thru Thursday.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor Peter Paras, can be reached on 571-272-4517. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/JANA A HINES/Primary Examiner, Art Unit 1645
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