The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
DETAILED ACTION
1. Preliminary amendment and claims 1-20 filed 12/12/24 are present and under consideration in this Office Action.
2. Priority
Receipt is acknowledged of papers (foreign priority filed 6/14/22 & 6/14/22) submitted under 35 U.S.C. 119(a)-(d), which papers have been placed of record in the file.
3. IDS filed 5/22/26 is considered. A signed copy of the IDS is provided with this Office Action.
4. Specification
The specification has not been checked to the extent necessary to determine the presence of all possible minor errors. Applicant's cooperation is requested in correcting any errors of which applicant may become aware in the specification.
5. Claim Rejections - 35 USC § 112 (second 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.
Claims 1-10 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 – step e. concentrating and drying the purified HMO-containing stream to obtain the neutral or sialylated HMO in solidified form, provided that the concentration step is optional when the drying step is freeze-drying.
The claim is confusing because if the “the neutral or sialylated HMO is already obtained in solidified form; the optional step is redundant. Also the use of the expression “optional” (claim 1), attempts to give both broad and narrow meaning to the scope of the above claims. The claim is unclear.
Claims 2-20 are included in the rejection for failing to correct the defect present in the base claim(s).
6. Claims 3, 4 & 18 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 3 recites “…wherein the active layer of the membrane is composed of polyamide and the MgSO.sub.4 rejection of the membrane is about 50-90%.”
The claim is unclear about the phrase “MgSO.sub.4 rejection of the membrane”. Clearly detailing what is meant by – “MgSO.sub.4 rejection of the membrane”, will overcome this rejection.
Claims 4 & 18 are included in the rejection for failing to correct the defect present in the base claim(s).
7. 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 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.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claim(s) 1-2, 4-10, 12 & 16 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by WO 2017/182,965 A1 (cited in the IDS).
Instant claims 1-20 are drawn to as follows:
1. A method for the purification of a neutral or sialylated human milk oligosaccharide (HMO) from a fermentation broth, comprising the steps of: a. separating the fermentation broth to form an HMO-containing stream and a biomass waste stream; b. purifying the HMO-containing stream by nanofiltration; c. purifying the HMO-containing stream with an acidic cation exchange resin; d. purifying the HMO-containing stream with an adsorbent resin; and e. concentrating and drying the purified HMO-containing stream to obtain the neutral or sialylated HMO in solidified form, provided that the concentration step is optional when the drying step is freeze-drying.
2. The method according to claim 1, wherein the nanofiltration membrane in step b) has a molecular weight cut-off (MWCO) of 500-3500 Da.
3. The method according to claim 2, wherein the active layer of the membrane is composed of polyamide and the MgSO.sub.4 rejection of the membrane is about 50-90%.
4. The method according to claim 3, wherein step b) is performed so that the pH is set below 5.0.
5. The method according to claim 1, wherein the acidic cation exchange resin in step c) is a strongly acidic cation exchange resin.
6. The method according to claim 5, wherein the strongly acidic cation exchange resin is a polystyrene-divinylbenzene cation exchange resin with sulfonic acid functional groups.
7. The method according to claim 1, wherein the concentration step before drying is evaporation or nanofiltration.
8. The method according to claim 1, wherein the drying step is spray-drying or freeze-drying.
9. The method according to claim 1, wherein the adsorbent resin is an acid adsorbent derived from cross-linked polystyrene or polyacrylic polymers and partially functionalized with tertiary amine functional groups.
10. The method according to claim 9, wherein the adsorbent resin has a surface area of >400 m.sup.2/g.
11. The method according to claim 10, wherein the adsorbent resin has an acid adsorbent capacity of 0.6-1.0 eq/kg on dry weight.
12. The method according to claim 1, wherein the HMO is a neutral HMO.
13. The method according to claim 12, wherein the neutral HMO is selected from the group consisting of: 2′-fucosyllactose, 3-fucosyllactose, 2′,3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-fucopentaose VI, lacto-N-difucohexaose I, lacto-N-difucohexaose II, lacto-N-difucohexaose III, 6′-galactosyllactose, 3′-galactosyllactose, lacto-N-hexaose and lacto-N-neohexaose.
14. The method according to claim 1, wherein the HMO is a sialylated HMO.
15. The method according to claim 14, wherein the sialylated HMO is 3′-sialyllactose (3′-SL) or 6′-sialyllactose (6′-SL).
16. The method according to claim 1, wherein the method lacks a purification/decolourization step with active carbon.
17. The method according to claim 12, wherein the neutral HMO is selected from the group consisting of: 2′-fucosyllactose, 3-fucosyllactose, 2′,3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose and a lacto-N-fucopentaose.
18. The method according to claim 3, wherein the polyamide is piperazine-based polyamide.
19. The method according to claim 6, wherein the polystyrene-divinylbenzene cation exchange resin with sulfonic acid functional groups is in H.sup.+-form.
20. The method according to claim 4, wherein step b) is performed so that the pH ranges from 3.0 to 4.5.
WO 2017/182,965 A1 teaches A method for separating an N-acetylglucosamine containing neutral oligosaccharide from dissolved inorganic and organic salts, acids and bases in an aqueous medium from a fermentation or enzymatic process, comprising the step of treating said aqueous medium with a strong cation exchange resin in H+-form and a weak anion exchange resin in free base form (See claim 1); which further comprises ultrafiltration and nanofiltration (See claim 4). The treatment with a strong cation exchange resin in H+-form and a weak anion exchange resin in free base form is preceded by ultrafiltration followed by nanofiltration (See claim 5). Ultrafiltration membrane can be used having a molecular weight cut-off (MWCO) range between about 1 and about 500 kDa, such as 10-250, 50-100, 200-500, 100-250, 1-100, 1-50, 10-25, 1-5 kDa, or any other suitable sub-ranges (page 15, line 12-22).
The application of a weak basic anion exchanger in free base form (resin's functional group is a tertiary amine) is advantageous (page 18, lines 9-17). Examples of a suitable basic anion exchange resin can be e.g. Amberlite IRA67, Amberlite IRA 96, Amberlite IRA743, Amberlite FPA53, Diaion CRB03, Diaion WA10, Dowex 66, Dowex Marathon, Lewatit MP64 (page 17, line 29 - page 18, line 3). As optional step, an active charcoal treatment is indicated. Spray drying and freeze drying is also indicated. Example 2 indicates a pH of 6,8. Evaporation can also be used (§ 2.2.4.1).
Also to enhance the purification of the N-acetylglucosamine containing neutral oligosaccharide, the pH of the aqueous medium is preferably adjusted to pH 3 to 8, such as 4 to 7, prior to reversed phase chromatography. See page 20, lines 14-16.
The adsorbed colour giving substances would still remain adsorbed on the charcoal, thus both decolourization and partial desalination can be achieved simultaneously in this optional step. See page 21, lines 20-22. Thus making the method lacks a purification/decolourization step with active carbon.
Applicants’ numerous claims limitations are also taught in WO 2017/182,965 – See for example claims 1-23. The N-acetylglucosamine containing core structures identified to date, for the 115 HMOs, are listed in Table 1. Lactose and the N-acetyllactosaminylated or lacto-N-biosylated core derivatives may further be substituted with one or more fucose and/or sialic acid residue(s), or lactose may be substituted with an additional galactose, to give HMOs known so far.
The invention relates to a method for obtaining an N-acetylglucosamine containing neutral oligosaccharide from a fermentation broth, wherein said oligosaccharide is produced by culturing a genetically modified microorganism capable of producing said oligosaccharide from an internalized carbohydrate precursor, comprising the steps of: i) ultrafiltration (UF), preferably to separate biomass from the broth, ii) nanofiltration (NF), preferably to concentrate said oligosaccharide in the broth and/or reduce an inorganic salt content of the broth, and iii) treating the broth with an ion exchange resin, preferably to remove charged materials, and/or subjecting the broth to chromatography, preferably to remove hydrophobic impurities. See the abstract of WO 2017/182,965.
8. Double Patenting Rejection
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the claims at issue are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); and In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on a nonstatutory double patenting ground provided the reference application or patent either is shown to be commonly owned with this application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP §§ 706.02(l)(1) - 706.02(l)(3) for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
Effective January 1, 1994, a registered attorney or agent of record may sign a terminal disclaimer. A terminal disclaimer signed by the assignee must fully comply with 37 CFR 3.73(b).
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/forms/. The filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to http://www.uspto.gov/patents/process/file/efs/guidance/eTD-info-I.jsp.
Claims 1-20 are provisionally rejected under the judicially created doctrine of double patenting over claims 1-20 of copending Application No.18/874209 (US 20250361256 A1). This is a provisional double patenting rejection since the conflicting claims have not yet been patented.
The subject matter claimed in the instant application is fully disclosed in the referenced copending application and would be covered by any patent granted on that copending application since the referenced copending application and the instant application are claiming common subject matter, as follows:
Claims 1-20 of the present application are listed above in paragraph 7. Claims of the copending application (not yet docketed) are drawn to claims 1-20 as follows.
1. A method for the purification of a neutral or sialylated human milk oligosaccharide (HMO) from a fermentation broth, comprising the steps of: I. separating an HMO-containing stream from biomass; II. purifying the HMO-containing stream with an adsorbent resin, and III. concentrating and drying the purified HMO-containing stream to obtain the neutral or sialylated HMOs in solidified form.
2. The method according to claim 1, the concentration step is optional when the drying step is freeze-drying.
3. The method according to claim 1, wherein the method does not comprise treatment with ion exchange resin.
4. The method according to claim 1, wherein the method does not comprise purification/decolourization step with active carbon.
5. The method according to claim 1, wherein the separated HMO-containing stream after step I) or the adsorbent resin eluate after step II) is purified by nanofiltration using a membrane having a molecular weight cut-off (MWCO) of 500-3500 Da, and the active (top) layer of the membrane is composed of polyamide.
6. The method according to claim 5, wherein step I) is ultrafiltration using a membrane having a MWCO of 500 Da to 5 kDa, wherein the active (top) layer of the membrane is not a polyamide material.
7. The method according to claim 5, wherein the nanofiltration membrane has an active layer of the membrane composed of piperazine-based polyamide, and its MgSO.sub.4 rejection is about 50-90%.
8. The method according to claim 7, wherein the nanofiltration step is performed so that the pH is set below 5.0.
9. The method according to claim 1, wherein the concentration step before drying is evaporation or nanofiltration.
10. The method according to claim 1, wherein the drying step is spray-drying or freeze-drying.
11. The method according to claim 1, wherein the adsorbent resin is an acid adsorbent derived from cross-linked polystyrene or polyacrylic polymers and partially functionalized with tertiary amine functional groups.
12. The method according to claim 11, wherein the adsorbent resin has a surface area of >400 m.sup.2/g, preferably the adsorbent resin has an acid adsorbent capacity of 0.6-1.0 eq/kg on dry weight.
13. The method according to claim 1, wherein the HMO is a neutral HMO.
14. The method according to claim 13, wherein the neutral HMO is selected from the group consisting of: 2′-fucosyllactose, 3-fucosyllactose, 2′,3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-fucopentaose VI, lacto-N-difucohexaose I, lacto-N-difucohexaose II, lacto-N-difucohexaose III, 6′-galactosyllactose, 3′-galactosyllactose, lacto-N-hexaose and lacto-N-neohexaose.
15. The method according to claim 1, wherein the HMO is a sialylated HMO.
16. The method according to claim 15, wherein the sialylated HMO is 3′-sialyllactose (3′-SL) or 6′-sialyllactose (6′-SL).
17. The method according to claim 13, wherein the neutral HMO is selected from the group consisting of: 2′-fucosyllactose, 3-fucosyllactose, 2′,3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose and a lacto-N-fucopentaose.
18. The method according to claim 13, wherein the neutral HMO is selected from the group consisting of: 2′-fucosyllactose, 3-fucosyllactose and 2′,3-difucosyllactose.
19. The method according to claim 8, wherein the pH is below 4.5.
20. The method according to claim 19, wherein the pH ranges from 3.0 to 4.5.
Given the fact pattern of the instant case as well as the copending application the species claims of the copending application anticipates the instant genus claims.
9. Claims 1-20 are provisionally rejected under the judicially created doctrine of double patenting over claims 1-20 of copending Application No.18/569,986 (US 20250128210 A1). This is a provisional double patenting rejection since the conflicting claims have not yet been patented.
Claims 1-20 of the present application are listed above in paragraph 7.
Claims of the copending application are drawn to claims 1-20 as follows.
1. A method for recovery and purification of a neutral or sialylated human milk oligosaccharide (HMO) from a fermentation broth, comprising the steps of: a) separating the fermentation broth to form a separated HMO-containing stream and a biomass waste stream; b) purifying the separated HMO-containing stream by nanofiltration; c) optionally concentrating the purified HMO-containing stream; and d) drying the purified HMO-containing stream to obtain a solidified HMO, wherein the nanofiltration membrane in step b) has a molecular weight cut-off (MWCO) of 500-3000 Da, the active layer of the membrane is composed of polyamide, and its MgSO.sub.4 rejection is about 50-90%, and which does not comprise an ion exchange resin treatment step or an electrodialysis step.
2. The method according to claim 1, wherein step a) comprises ultrafiltration or centrifugation.
3. The method according to claim 1, wherein step b) is performed so that the pH is set below 5.0.
4. The method according to claim 1, wherein step b) further comprises nanofiltration conducted in diafiltration mode.
5. The method according to claim 1, wherein step b) comprises a second nanofiltration step, wherein the nanofiltration membrane has a molecular weight cut-off (MWCO) of 150-300 Da.
6. The method according to claim 1, wherein step c) comprises evaporation, nanofiltration, reverse-osmosis filtration, or a combination thereof.
7. The method according to claim 1, wherein step c) comprises concentration with a nanofiltration membrane, wherein the nanofiltration membrane has a molecular weight cut-off (MWCO) of 150-300 Da.
8. The method according to claim 1, wherein step c) comprises concentration with a nanofiltration membrane, the nanofiltration membrane is in the range of 500-3000 Da MWCO, has an active (top) layer composed of piperazine-based polyamide, a MgSO4 rejection factor of about 50-90% and a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0.
9. The method according to claim 1, wherein step c) is conducted and step d) consists of spray-drying to obtain solidified HMO.
10. The method according to claim 1, further comprising an active carbon treatment step following step b) or step c).
11. The method according to claim 1 comprising the following steps: i. separating the fermentation broth to form a separated neutral or sialylated HMO-containing stream and a biomass waste stream by ultrafiltration; ii. purifying the separated neutral or sialylated HMO-containing stream by combined nanofiltration and diafiltration; iii. purifying the neutral or sialylated HMO-containing stream by active carbon treatment; iv. purifying the separated neutral or sialylated HMO-containing stream by a second nanofiltration step; v. concentrating the purified neutral or sialylated HMO-containing stream by evaporation; and vi. spray-drying the purified neutral or sialylated HMO-containing stream to obtain solidified neutral or sialylated HMO.
12. The method according to claim 1 comprising the following steps: i. separating the fermentation broth to form a separated neutral or sialylated HMO-containing stream and a biomass waste stream by ultrafiltration; ii. purifying the separated neutral or sialylated HMO-containing stream by combined nanofiltration and diafiltration; iii. purifying the neutral or sialylated HMO-containing stream by active carbon treatment; iv. optionally purifying the separated neutral or sialylated HMO-containing stream by a second nanofiltration step, optionally combined with diafiltration; v. concentrating the purified neutral or sialylated HMO-containing stream by evaporation; and vi. spray-drying the purified neutral or sialylated HMO-containing stream to obtain solidified neutral or sialylated HMO.
13. The method according to claim 11, wherein the nanofiltration membrane has an active (top) layer composed of polyamide, the membrane has a MgSO.sub.4 rejection factor of about 50-90% and a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0.
14. The method according to claim 1 comprising the following steps: i. separating the fermentation broth to form a separated neutral or sialylated HMO-containing stream and a biomass waste stream by ultrafiltration; ii. purifying the separated neutral or sialylated HMO-containing stream by combined nanofiltration and diafiltration; iii. purifying the neutral or sialylated HMO-containing stream by active carbon treatment; iv. optionally purifying the separated neutral or sialylated HMO-containing stream by a second nanofiltration step, optionally combined with diafiltration, wherein the nanofiltration membrane is in the range of 500-3000 Da MWCO, has an active (top) layer composed of polyamide, a MgSO.sub.4 rejection factor of about 50-90% and a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0; v. concentrating the purified neutral or sialylated HMO-containing stream by evaporation; and vi. spray-drying the purified neutral or sialylated HMO-containing stream to obtain solidified neutral or sialylated HMO.
15. The method according to claim 1, wherein the HMO is a neutral HMO.
16. The method according to claim 15, wherein the HMO is selected from the group consisting of: 2′-fucosyllactose, 3-fucosyllactose, 2′,3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-fucopentaose VI, lacto-N-difucohexaose I, lacto-N-difucohexaose II, lacto-N-difucohexaose III, 6′-galactosyllactose, 3′-galactosyllactose, lacto-N-hexaose and lacto-N-neohexaose.
17. The method according to 15, wherein the HMO is 2′-fucosyllactose, 3-fucosyllactose, 2′,3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose or a lacto-N-fucopentaose.
18. The method according to claim 1, wherein the HMO is a sialylated HMO.
19. The method according to claim 3, wherein the pH is set to between 3.0 and 4.5.
20. The method according to claim 13, wherein the pH is set to between 3.0 and 4.5.
Given the fact pattern of the instant case as well as the copending application the narrower instant claims are obvious variation of the claims in the copending application.
10. Claims 1-20 are provisionally rejected under the judicially created doctrine of double patenting over claims 1-20 of copending Application No.18/570,003 (US 20240286081 A1). This is a provisional double patenting rejection since the conflicting claims have not yet been patented.
Claims 1-20 of the present application are listed above in paragraph 7.
Claims 1-19 & 21 of the copending application as follows:
1. A method for recovery and purification of a neutral or sialylated human milk oligosaccharide (HMO) from a fermentation broth, comprising the steps of: a. separating the fermentation broth to form a separated HMO-containing stream and a biomass waste stream; b. purifying the separated HMO-containing stream by membrane filtration using a membrane having a MWCO of 500 Da to 5 kDa, wherein the active (top) layer of the membrane is not a polyamide material, and wherein the rejection factor for the HMO with respect to the membrane is less than 90% allowing at least a part of the HMO product into permeate with optional diafiltration so that the HMO accumulates in the permeate and ensuring the retention of impurities with higher molecular weight than that of the HMO; c. purifying the HMO-containing stream by nanofiltration; d. optionally concentrating the purified HMO-containing stream; and e. drying the purified HMO-containing stream to obtain a solidified neutral or sialylated HMO. wherein the method does not comprise a basic anion exchange resin treatment step and/or an electrodialysis step.
2. The method according to claim 1, wherein the method does not comprise an ion exchange resin treatment step or an electrodialysis step.
3. The method according to claim 1, wherein step a) comprises ultrafiltration with an ultrafiltration membrane having a molecular weight cut-off (MWCO) higher than 30 kDa and lower than 500 kDa and/or centrifugation.
4. The method according to claim 1, wherein the nanofiltration membrane in step c) has a molecular weight cut-off (MWCO) of 500-3000 Da, and the active layer of the membrane is composed of polyamide and its MgSO.sub.4 rejection is about 50-90%.
5. The method according to claim 4, wherein step c) is performed so that the pH is set below 5.0.
6. The method according to claim 1, wherein step c) further comprises nanofiltration conducted in diafiltration mode.
7. The method according to claim 1, wherein the method comprises further purification of the HMO-containing stream with a cation exchange resin.
8. The method according to claim 1, wherein the method comprises further purification of the HMO-containing stream by an active carbon treatment.
9. The method according to claim 1, wherein step d) comprises evaporation, nanofiltration, reverse-osmosis filtration, or a combination thereof.
10. The method according to claim 1, wherein step d) comprises concentration with a nanofiltration membrane with a molecular weight cut-off (MWCO) of 150-300 Da.
11. The method according to claim 1, wherein step d) comprises concentration with a nanofiltration membrane, the nanofiltration membrane is in the range of 500-3000 Da MWCO, has an active (top) layer composed of piperazine-based polyamide, a MgSO.sub.4 rejection factor of about 50-90% and a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0.
12. The method according to claim 1, wherein step d) is conducted and step e) consists of spray-drying to obtain solidified HMO.
13. The method according to claim 1, further comprising an active carbon treatment step following step c) or step d).
14. The method according to claim 1 comprising the following steps: i. separating the fermentation broth to form a separated HMO-containing stream and a biomass waste stream by ultrafiltration using an ultrafiltration membrane having a MWCO of higher than 10 kDa and lower than 500 kDa; ii. purifying the separated HMO-containing stream by membrane filtration using a membrane having a MWCO of 500 Da to 5 kDa, wherein the active (top) layer of the membrane is not a polyamide material, and wherein the rejection factor for the HMO with respect to the membrane is less than 90% allowing at least a part of the HMO product into permeate with optional diafiltration so that the HMO accumulates in the permeate and ensuring the retention of impurities with higher molecular weight than that of the HMO; iii. purifying the separated neutral or sialylated HMO-containing stream by combined nanofiltration and diafiltration; iv. purifying the neutral or sialylated HMO-containing stream by active carbon treatment; v. optionally purifying the separated neutral or sialylated HMO-containing stream by a second nanofiltration step, optionally combined with diafiltration, wherein the nanofiltration membrane is in the range of 500-3000 Da MWCO, has an active (top) layer composed of polyamide, a MgSO.sub.4 rejection factor of about 50-90% and preferably-a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0; vi. concentrating the purified neutral or sialylated HMO-containing stream by evaporation; and vii. spray-drying the purified neutral or sialylated HMO-containing stream to obtain solidified neutral or sialylated HMO.
15. The method according to claim 1 comprising the following steps: i. separating the fermentation broth to form a separated HMO-containing stream and a biomass waste stream by ultrafiltration using an ultrafiltration membrane having a MWCO of higher than 10 kDa and lower than 500 kDa; ii. purifying the separated HMO-containing stream by membrane filtration using a membrane having a MWCO of 500 Da to 5 kDa, wherein the active (top) layer of the membrane is not a polyamide material, and wherein the rejection factor for the HMO with respect to the membrane is less than 90% allowing at least a part of the HMO product into permeate with optional diafiltration so that the HMO accumulates in the permeate and ensuring the retention of impurities with higher molecular weight than that of the HMO; iii. purifying the HMO-containing stream by combined nanofiltration and diafiltration; iv. purifying the separated neutral or sialylated HMO-containing stream by diafiltration; v. purifying the neutral or sialylated HMO-containing stream by active carbon treatment; vi. optionally purifying the separated neutral or sialylated HMO-containing stream by a second nanofiltration step, wherein the nanofiltration membrane is in the range of 150-300 Da MWCO; vii. optionally purifying the separated neutral or sialylated HMO-containing stream by a second diafiltration step; viii. concentrating the purified neutral or sialylated HMO-containing stream by evaporation; and ix. spray-drying the purified neutral or sialylated HMO-containing stream to obtain solidified neutral or sialylated HMO.
16. The method according to claim 14, wherein the nanofiltration membrane has an active (top) layer composed of piperazine-based polyamide, the membrane has a MgSO.sub.4 rejection factor of about 50-90% and a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0.
17. The method according to claim 1 comprising the following steps: i. separating the fermentation broth to form a separated HMO-containing stream and a biomass waste stream by ultrafiltration using an ultrafiltration membrane having a MWCO of higher than 10 kDa and lower than 500 kDa; ii. purifying the separated HMO-containing stream by membrane filtration using a membrane having a MWCO of 500 Da to 5 kDa, wherein the active (top) layer of the membrane is not a polyamide material, and wherein the rejection factor for the HMO with respect to the membrane is less than 90% allowing at least a part of the HMO product into permeate with optional diafiltration so that the HMO accumulates in the permeate and ensuring the retention of impurities with higher molecular weight than that of the HMO; iii. purifying the separated neutral or sialylated HMO-containing stream by combined nanofiltration and diafiltration; iv. purifying the neutral or sialylated HMO-containing stream by active carbon treatment; v. optionally purifying the separated neutral or sialylated HMO-containing stream by a second nanofiltration step, optionally combined with diafiltration, wherein the nanofiltration membrane has an active (top) layer composed of piperazine-based polyamide, the membrane has a MgSO.sub.4 rejection factor of about 50-90% and a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0; vi. concentrating the purified neutral or sialylated HMO-containing stream by evaporation; and vii. spray-drying the purified neutral or sialylated HMO-containing stream to obtain solidified neutral or sialylated HMO.
18. The method according to claim 1, wherein the HMO is a neutral HMO.
19. The method according to claim 18, wherein the HMO is selected from the group consisting of: 2′-fucosyllactose, 3-fucosyllactose, 2′,3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-fucopentaose VI, lacto-N-difucohexaose I, lacto-N-difucohexaose II, lacto-N-difucohexaose III, 6′-galactosyllactose, 3′-galactosyllactose, lacto-N-hexaose and lacto-N-neohexaose.
20. (canceled)
21. The method according to claim 1, wherein the HMO is a sialylated HMO.
Given the fact pattern of the instant case as well as the copending application the narrower instant claims are obvious variation of the claims in the copending application.
11. Claims 1-20 are provisionally rejected under the judicially created doctrine of double patenting over claims 1-20 of copending Application No.18/570,021 (US20240287116 A1). This is a provisional double patenting rejection since the conflicting claims have not yet been patented.
Claims 1-20 of the present application are listed above in paragraph 7.
Claims 1-20 of the copending application as follows:
1. A method for recovery and purification of a neutral or sialylated human milk oligosaccharide (HMO) from a fermentation broth, comprising the steps of: I. separating the fermentation broth to form a separated HMO-containing stream and a biomass waste stream; IIa. purifying the separated HMO-containing stream by nanofiltration (NF) or nanofiltration/diafiltration (NF/DF); IIb. an acidic cation exchange resin treatment then a nanofiltration step, wherein the nanofiltration membrane has a molecular weight cut-off (MWCO) of 500-3000 Da, the active (top) layer composed of polyamide, the membrane has a MgSO.sub.4 rejection factor of about 50-90% and a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0; III. optionally concentrating the purified HMO-containing stream; and IV. drying the purified HMO-containing stream to obtain a solidified neutral or sialylated HMO, with optional active charcoal treatment.
2. The method according to claim 1, wherein step I) comprises at least one of ultrafiltration, microfiltration, and centrifugation.
3. The method according to claim 1, wherein the acidic cation exchange resin in step IIb) is a strongly acidic cation exchange resin, preferably a styrene-divinylbenzene cation exchange resin.
4. The method according to claim 3, wherein the resin is in H.sup.+-form.
5. The method according to claim 1, wherein the method comprises further purification of the HMO-containing stream by an active carbon treatment.
6. The method according to claim 1, wherein step IIa) or IIb) further comprises nanofiltration conducted in diafiltration mode.
7. The method according to claim 1, wherein step III) comprises evaporation, reverse-osmosis filtration, or a combination thereof.
8. The method according to claim 1, wherein step III) comprises concentration with a nanofiltration membrane, wherein the nanofiltration membrane has a molecular weight cut-off (MWCO) of 150-300 Da.
9. The method according to claim 1, wherein step III) is conducted and step IV) consists of spray-drying to obtain solidified HMO.
10. The method according to claim 1 comprising the following steps: i. separating the fermentation broth to form a separated HMO-containing stream and a biomass waste stream by ultrafiltration; ii. purifying the separated HMO-containing stream by combined nanofiltration and diafiltration, wherein the nanofiltration membrane is in the range of 500-3000 Da MWCO; iii. purifying the HMO-containing stream by a strongly acidic cation exchange resin in H.sup.+-form; iv. purifying the HMO-containing stream by a second nanofiltration step, wherein the nanofiltration membrane has a molecular weight cut-off (MWCO) of 500-3000 Da, the active (top) layer composed of piperazine-based polyamide, the membrane has a MgSO.sub.4 rejection factor of about 50-90% and a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0; v. concentrating the purified HMO-containing stream by evaporation; and vi. spray-drying the purified HMO-containing stream to obtain a solidified neutral or sialylated HMO; optionally with active charcoal treatment after step ii), iii), iv) or v).
11. The method according to claim 1 comprising the following steps: i. separating the fermentation broth to form a separated HMO-containing stream and a biomass waste stream by ultrafiltration; ii. purifying the separated HMO-containing stream by combined nanofiltration and diafiltration, wherein the nanofiltration membrane is in the range of 500-3000 Da MWCO; iii. purifying the nanofiltration retentate by a strongly acidic cation exchange resin in H.sup.+-form; iv. purifying the resin eluate by a second nanofiltration step, preferably combined with diafiltration, wherein the nanofiltration membrane has a molecular weight cut-off (MWCO) of 500-3000 Da, the active (top) layer composed of piperazine-based polyamide, the membrane has a MgSO.sub.4 rejection factor of about 50-90% and a NaCl rejection factor of not more than 50%, and the nanofiltration step is performed so that the pH is set below 5.0; v. optionally concentrating the nanofiltration retentate by evaporation or reverse osmosis; and vi. freeze-drying the nanofiltration retentate or the concentrate to obtain a solidified neutral or sialylated HMO; optionally with active charcoal treatment between steps iii) and iv).
12. The method according to claim 10, wherein step iii) further comprises the addition of NaOH-solution to the resin eluate so that the pH is set to 3-5.
13. The method according to claim 1 which does not comprise a basic anion exchange resin treatment.
14. The method according to claim 13 which does not comprise electrodialysis.
15. The method according to claim 1, wherein the HMO is a neutral HMO.
16. The method according to claim 15, wherein the HMO is selected from the group consisting of: 2′-fucosyllactose, 3-fucosyllactose, 2′,3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-fucopentaose VI, lacto-N-difucohexaose I, lacto-N-difucohexaose II, lacto-N-difucohexaose III, 6′-galactosyllactose, 3′-galactosyllactose, lacto-N-hexaose and lacto-N-neohexaose.
17. The method according to 15, wherein the HMO is 2′-fucosyllactose, 3-fucosyllactose, 2′,3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose or a lacto-N-fucopentaose, preferably 2′-fucosyllactose, LNT, LNnT or a lacto-N-fucopentaose.
18. The method according to claim 1, wherein the HMO is a sialylated HMO.
19. The method according to claim 1, wherein the pH is set between 3.0 and 4.5.
20. The method according to claim 10, wherein the pH is set between 3.0 and 4.5.
Given the fact pattern of the instant case as well as the copending application the narrower instant claims are obvious variation of the claims in the copending application.
12. No claim is allowed.
13. Any inquiry concerning this communication or earlier communications from the examiner should be directed to TEKCHAND SAIDHA whose telephone number is (571)272-0940. The examiner can normally be reached on M-F 8.00-5.30. 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.
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/TEKCHAND SAIDHA/
Primary Examiner, Art Unit 1652
Recombinant Enzymes, Hoteling
Telephone: (571) 272-0940
Fax: (571) 273-0940
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