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CN-121985886-A - Hollow fiber

CN121985886ACN 121985886 ACN121985886 ACN 121985886ACN-121985886-A

Abstract

The present disclosure provides extruded or spun semipermeable porous hollow fibers comprising covalent ester, thioester, and/or amide crosslinked polypeptides and methods of producing the same. The hollow fibers may be produced from proteins, protein extracts and/or protein isolates derived from plants, animals, bacteria, algae, archaea and/or fungi, and in certain embodiments are intended to be suitable for human and/or animal ingestion. In some embodiments, the hollow fibers may be designed for producing cartridges compatible with existing and/or new bioreactor platforms for containing cell cultures in the production of cultured meat.

Inventors

  • R.J. MARTIN
  • H. R. Ingram

Assignees

  • 知识产权风险投资股份有限公司

Dates

Publication Date
20260505
Application Date
20240402
Priority Date
20230406

Claims (20)

  1. 1. An edible semipermeable porous hollow fiber comprising one or more polycarboxylic acid derived covalent ester, thioester and/or amide bond crosslinked polypeptides, Wherein: (a) The outer diameter of the hollow fiber is 50-6600 mu M; (b) The wall thickness of the hollow fiber is 20-800 mu M; (c) The hollow fiber has a lumen diameter of 20-5000 μm, and (D) The hollow fiber has a pore volume of 1% -95%.
  2. 2. A semipermeable porous hollow fiber comprising one or more polycarboxylic acid-derived covalent ester, thioester and/or amide bond crosslinked polypeptides, Wherein: (a) The outer diameter of the hollow fiber is 50-6600 mu M; (b) The wall thickness of the hollow fiber is 20-800 mu M; (c) The hollow fiber has a lumen diameter of 20-5000 μm, and (D) The hollow fiber has a pore volume of 1% -95%.
  3. 3. An edible semipermeable porous hollow fiber comprising one or more polycarboxylic acid derived covalent ester, thioester and/or amide bond crosslinked polypeptides, Wherein: (a) The outer diameter of the hollow fiber is 50-6600 mu m; (b) The wall thickness of the hollow fiber is 20-800 mu m; (c) The hollow fiber has a lumen diameter of 20-5000 μm, and (D) The porosity of the hollow fiber is 1% -95%.
  4. 4. A semipermeable porous hollow fiber comprising one or more polycarboxylic acid-derived covalent ester, thioester and/or amide bond crosslinked polypeptides, Wherein: (a) The outer diameter of the hollow fiber is 50-6600 mu m; (b) The wall thickness of the hollow fiber is 20-800 mu m; (c) The hollow fiber has a lumen diameter of 20-5000 μm, and (D) The porosity of the hollow fiber is 1% -95%.
  5. 5. A method for producing a semipermeable porous hollow fiber comprising a covalently crosslinked polypeptide, the method comprising the steps of: a. Combination: i. a first composition comprising a polypeptide, and A second composition comprising a solvent and one or more denaturants and/or reducing agents to produce a third composition; b. Incubating the third composition under conditions sufficient to solubilize, denature and/or reduce at least a portion of the polypeptide, and To produce a fourth composition; c. extruding or spinning the fourth composition with a pore solution through a plurality of coaxial orifices to produce hollow fibers; d. treating the hollow fibers with a polycarboxylic acid crosslinking reagent to form covalent cross-links of the polypeptide and/or polycarboxylic acid-derived esters, thioesters, or amides within the polypeptide within at least a portion of the polypeptide in the hollow fibers to produce covalently cross-linked semipermeable porous hollow fiber hollow fibers; e. Treating the covalently crosslinked semipermeable porous hollow fibers with one or more post-production modification methods selected from the group consisting of: i. treating the covalently cross-linked semipermeable porous hollow fiber with an organic solvent to increase the relative abundance of beta-sheets in the secondary structure of the polypeptide, and Annealing the covalently crosslinked semipermeable porous hollow fibers; Treating the covalently crosslinked semipermeable porous hollow fibers with a solvent to remove void-containing elements in the hollow fibers; Washing the covalently crosslinked semipermeable porous hollow fibers with one or more acids, bases and/or buffer solutions to reduce at least one of young's modulus, ultimate tensile strength and/or ultimate tensile strain of the hollow fibers; v. coating the covalently crosslinked semipermeable porous hollow fibers; modifying the surface morphology of the covalently crosslinked semipermeable porous hollow fibers to aid cell attachment and/or cell alignment; To produce a plurality of treated covalently cross-linked semipermeable porous hollow fiber hollow fibers, and F. Drying the treated covalently crosslinked semipermeable porous hollow fibers to produce dried treated covalently crosslinked semipermeable porous hollow fibers.
  6. 6. The method of claim 5, further providing that the first composition comprises a plant-derived polypeptide.
  7. 7. The method of claims 5 to 6, further providing for the supplementation of polysaccharides to the first composition of step (a) of claim 5 in a proportion in the range of 1% to 10000% (weight/weight of polypeptide).
  8. 8. The method of any one of claims 5 to 7, further providing for the supplementation of lipids into the first composition of step (a) of claim 5 in a proportion in the range of 1% to 10000% (weight/weight of polypeptide).
  9. 9. The method of any one of claims 5 to 8, further providing for supplementing a polyol and/or a polymer of a polyol into the first composition of step (a) of claim 5 in a proportion in the range of 1% to 10000% (weight/weight of polypeptide).
  10. 10. The method of any one of claims 5 to 9, wherein the second composition consists of one or more solvents comprising water, ethanol, oxalic acid, malic acid, succinic acid, adipic acid, tartaric acid, citric acid, malonic acid, acetic acid, formic acid, sodium hydroxide, and/or potassium hydroxide.
  11. 11. The method of any one of claims 5 to 10, wherein the second composition comprises one or more solvents comprising water, ethanol, oxalic acid, malic acid, succinic acid, adipic acid, tartaric acid, citric acid, malonic acid, acetic acid, formic acid, sodium hydroxide, and/or potassium hydroxide.
  12. 12. The method of any one of claims 5 to 11, wherein the second composition comprises one or more denaturants that are at least one of oxalic acid, malic acid, succinic acid, adipic acid, tartaric acid, citric acid, malonic acid, acetic acid, formic acid, urea, sodium hydroxide, and/or potassium hydroxide.
  13. 13. The method of any one of claims 5 to 12, wherein the second composition consists of one or more reducing agents comprising N-acetylcysteine, L-cysteine, glutathione, ascorbic acid, citric acid, tartaric acid, malic acid, sodium borohydride, sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium hypophosphite, sodium dithionite, mercaptoethanol, and/or dithiothreitol.
  14. 14. The method of any one of claims 5 to 13, wherein the second composition comprises one or more reducing agents comprising N-acetylcysteine, L-cysteine, glutathione, ascorbic acid, citric acid, tartaric acid, malic acid, sodium borohydride, sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium hypophosphite, sodium dithionite, mercaptoethanol, and/or dithiothreitol.
  15. 15. The method of any one of claims 5 to 14, wherein the third composition of step (b) is incubated for a total duration of 5 minutes to 1 week.
  16. 16. The method of any one of claims 5 to 15, wherein the third composition or the fourth composition of step (b) is incubated at a temperature between 0 ℃ for a minute and 150 ℃.
  17. 17. The method of any one of claims 5 to 16, wherein the fourth composition comprises one or more void elements comprising calcium carbonate, ice, air, nitrogen, carbon dioxide, argon, sodium chloride, or potassium chloride.
  18. 18. The method of any one of claims 5 to 17, further providing for adding a void-containing element to the composition in step (a) or step (b) such that the void-containing element comprises from 1% to 80% of the fourth composition (iv).
  19. 19. The method of any one of claims 5 to 18, wherein the coagulation bath solution comprises one or more polycarboxylates comprising sodium citrate, sodium malate, potassium malate, and/or potassium citrate.
  20. 20. The method of any one of claims 5 to 19, further providing for adding a salt to the composition of the coagulation bath solution, the salt comprising zinc sulfate, sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium hypophosphite, and/or ammonium sulfate.

Description

Hollow fiber Technical Field In certain aspects and embodiments, the present disclosure relates in part to hollow fibers and methods of producing and using the same. 1.2 Abstract In some aspects and embodiments of the present disclosure, there are provided extruded or spun semipermeable porous hollow fibers comprising covalent ester, thioester, and/or amide crosslinked polypeptides and methods of producing the same. The hollow fibers of the present disclosure may be produced from proteins, protein extracts, and/or protein isolates derived from plants, animals, bacteria, algae, archaea, and/or fungi, and are intended in certain embodiments to be suitable for human and/or animal ingestion. In some embodiments, the hollow fibers of the present disclosure may be designed for use in producing cartridges compatible with existing and/or new bioreactor platforms for containing cell cultures in cultured meat production. Background U.S. patent application publication No. US2013/0192459 discloses "hollow fiber membranes having a support layer and an inner (i.e., lumen-facing side of the support layer) or outer separation layer, and methods for manufacturing such hollow fiber membranes". PCT application No. WO2019158494A1 discloses "an edible fiber comprising a biopolymer and a plasticizer, wherein the weight ratio of biopolymer to plasticizer is from about 1:0.25 to about 1:3, and wherein the fiber has a diameter of from about 0.5 μm to about 1 mm". European patent application No. EP0077098A2 discloses "hollow chitosan fibers in general ultrafiltration and dialysis processes (and in particular renal dialysis) and a process for their preparation". WO 2016/007879A1 discloses that "a bioprotein tube having an outer diameter of about 1mm and an inner diameter of about 0.80 can be prepared by extruding a bioprotein precursor solution comprising 50mg/ml alginate and gelatin (in a 3:1 ratio) through an orifice into a solution of about 5mg/ml calcium chloride and exposing the tube to a wash solution within about 10 seconds". WO 2018/01805 A2 discloses "a system for culturing cells that may be used to produce edible meat in some embodiments of the invention". It also indicates that "cultivation is performed on edible hollow fiber cartridges". In addition, "plant derived substrate is from the families of the cereal, leguminous (Fabaceae) or pseudocereal families. In addition, "beans are soybeans or peas". WO 2022/038241A1 discloses "a closed, continuous, semi-continuous or batch culture system for cell growth and differentiation followed by tissue growth to produce, for example, clean meat. " WO 2009/035414A1 discloses "the present invention relates to a chitosan construct and a method for preparing the same". The chitosan construct may have a high mechanical strength. In particular, the present invention relates to chitosan fibers or chitosan hollow fibers. It also indicates that "in particular, the construct may have a spongy porous structure. The porosity of the construct can be as high as 80%. For example, the porosity may be about 20% -80% ". WO 2022/038240A2 discloses "a method for producing structured clean meat products produced with hollow fibers, cartridges and bioreactors". WO 2023/021213A1 discloses "methods of making crosslinked, edible, porous hollow fibers and sheet-like films suitable for making clean meat products, hollow fibers and sheet-like films made therefrom, and methods of using the same. Among them, "the innovation of the present invention is the use of physical crosslinking that is generated via an energy source, such as one or more of heat, gamma, electron beam, beta, x-ray, or UV. Modrzejewska and Eckstein, biopolymers,73:61-68 (2004) concluded that "chitosan hollow fibers can be formed by wet weaving using a water-solidifying agent (sodium hydroxide). JP2022072917a discloses "a cultured meat complex comprising a hollow fiber membrane or its decomposition product or lysate and a cultured meat of an animal cell population present along the hollow fiber membrane or its decomposition product or lysate". Among them, "a cultured meat composite obtained using a hollow fiber membrane composed of collagen may be subjected to a heat treatment to gel the collagen". WO 2023/152492A1 discloses "a matrix assembly for culturing cells, wherein the matrix assembly comprises a plurality of edible fibers, wherein each fiber has an internal channel extending along its length. Wherein the "plurality of fibers may comprise alginate. Wherein "calcium chloride acts as a crosslinker for the alginate. WO 2023/152493A1 discloses "a plurality of edible fibers, wherein each fiber has an internal channel extending along its length". Wherein "calcium chloride is used as a cross-linking agent for the alginate" and "freeze-drying step" can also be used to adjust the porosity of the final fibrous material. Furthermore, "it has been found that the freeze-drying step is particularly useful when the fibers comprise alginate. WO 2024/03