Method for the manufacture of recombinant unhydroxylated collagen polypeptide fibres, and recombinant unhydroxylated collagen polypeptide fibres obtained thereby
The present invention relates to a process for the production of fibres of an unhydroxylated recombinant collagen polypeptide, and the fibres obtained thereby, as well as to uses of such fibres as a biomaterial.
1. A method for producing an unhydroxylated recombinant collagen polypeptide fiber comprising:
a. diluting an unhydroxylated recombinant collagen polypeptide in an acid at a temperature of between about 4° C. and about 26° C.; and
b. dialyzing the diluted unhydroxylated recombinant collagen polypeptide against a buffer below the usual physiological temperature of the corresponding native collagen to form an unhydroxylated recombinant collagen polypeptide fiber.
2. The method of claim 1 , wherein the unhydroxylated recombinant collagen polypeptide is expressed by a plant.
3. The method of claim 1 , wherein the unhydroxylated recombinant collagen polypeptide expressed by a plant is obtained by insertion of one or more DNA sequences coding for said polypeptide into a plant.
4. The method of claim 1 , wherein the recombinant collagen polypeptide is a collagen comprising at least one human alpha I chain.
5. The method of claim 4 , wherein the unhydroxylated recombinant collagen polypeptide is a collagen comprising at least one human alpha I chain, and at least another human alpha chain.
6. The method of claim 4 , wherein the unhydroxylated recombinant collagen polypeptide is a collagen comprising at least three human alpha I chains.
7. The method of claim 1 , wherein the acid is 0.1M acetic acid.
8. The method of claim 1 , wherein the dialysis is carried out at 4° C.
9. The method of claim 1 , wherein the dialysis is carried out against a monopotassium phosphate buffer without the addition of salts.
10. The method of claim 9 , wherein the dialysis is carried out against a 10 mM pH 7 monopotassium phosphate buffer for 12 hours.
11. The method of claim 1 , wherein the fibers formed after dialysis are separated by centrifugation.
12. The method of claim 11 , wherein centrifugation is carried out at about 20000 G for about 10 minutes.
13. The method of claim 1 , wherein the fibers have a diameter of between about 30 nm to about 550 nm.
14. The method of claim 1 , further comprising the step of reticulating said unhydroxylated recombinant collagen fiber.
15. The method of claim 1 , further comprising the step of incorporating said unhydroxylated recombinant collagen fiber into a biomaterial.
16. The method of claim 15 , wherein said biomaterial is in the form of a sponge.
17. The method of claim 15 , wherein said biomaterial is in the form of a film or a gel.
18. The method of claim 17 , wherein said film or gel adheres to human tissue.
19. The method of claim 17 , wherein said film or gel does not adhere to human tissue.
20. The method of claim 15 , wherein said biomaterial has a three dimensional structure.
21. The method of claim 20 , wherein said three dimensional structure is the form of a tube.
22. The method of claim 15 , wherein said biomaterial is incorporated by gluing, spraying, induction, impregnation, fusion or extrusion to a prosthetic structure.
23. The method of claim 21 , wherein said prosthetic structure is a knitted web.
24. The method of claim 23 , wherein said prosthetic structure is an unknitted web.
25. The method of claim 21 , wherein said prosthetic structure is a vascular prosthesis.
26. The method of claim 23 , wherein said prosthetic structure is an endovascular prosthesis.
27. The method of claim 26 , wherein said endovascular prosthesis is a stent.