Regeneration of chromatography material
View Patent ↗Methods and kits for regenerating antibody binding resins are provided. In some embodiments the methods include a first step of washing an antibody binding resin with a reducing solution, followed by a second step of washing the antibody binding resin with a chaotropic solution.
1. A method for regenerating an antibody binding resin, the method comprising:
a first step of washing an antibody binding resin with a reducing solution, followed by a second step of washing the antibody binding resin with a chaotropic solution, thereby regenerating the antibody binding resin, wherein the first step and the second step are performed separately.
2. The method of claim 1 , further comprising a step of washing the antibody binding resin with an equilibration buffer, wherein the step of washing with the equilibration buffer is a separate step that is performed after the first washing step and prior to the second washing step.
3. The method of claim 2 , wherein the equilibration buffer is selected from the group consisting of phosphate buffers and carbonate buffers.
4. The method of claim 3 , wherein the equilibration buffer is a phosphate buffer.
5. The method of claim 1 , further comprising washing the antibody binding resin with an acidic solution, wherein the step of washing with the acidic solution is a separate step that is performed prior to the first washing step.
6. The method of claim 5 , wherein the acidic solution is selected from the group consisting of phosphoric acid and acidic acid.
7. The method of claim 6 , wherein the acidic solution is phosphoric acid.
8. The method of claim 5 , further comprising washing the antibody binding resin with a neutralizing solution, wherein the step of washing with the neutralizing solution is a separate step that is performed after the step of washing with the acidic solution and prior to the first washing step.
9. The method of claim 8 , wherein the neutralizing solution is selected from the group consisting of phosphate buffers, weak acids and weak bases.
10. The method of claim 9 , wherein the neutralizing solution is a phosphate buffer.
11. The method of claim 1 , further comprising using the antibody binding resin to purify an antibody.
12. The method of claim 1 , further comprising repeating the first and second washing steps.
13. The method of claim 1 , wherein the reducing solution includes a reducing agent selected from the group consisting of thioglycerol, 1-4,-dithiothreitol, and 2-mercaptoethanol.
14. The method of claim 13 , wherein the reducing agent is thioglycerol.
15. The method of claim 1 , wherein the chaotropic solution includes a chaotropic agent selected from the group consisting of guanidine HCl, urea, and guanidine acetate.
16. The method of claim 15 , wherein the chaotropic agent is guanidine HCl or urea.
17. The method of claim 1 , wherein the antibody binding resin is selected from the group consisting of a protein A resin and a protein G resin.
18. The method of claim 17 , wherein the antibody binding resin is a protein A resin.
19. A method for regenerating an antibody binding resin, the method comprising the following ordered steps:
(a) a first step of washing an antibody binding resin with a reducing solution selected from the group consisting of thioglycerol, 1-4,-dithiothreitol, and 2-mercaptoethanol;
(b) a second step of washing the antibody binding resin with an equilibration buffer selected from the group consisting of phosphate buffers and carbonate buffers; and
(c) a third step of washing the antibody binding resin with a chaotropic solution selected from the group consisting of guanidine HCl, urea, and guanidine acetate, thereby regenerating the antibody binding resin.
20. The method of claim 19 , wherein the reducing agent is thioglycerol.
21. The method of claim 19 , wherein the chaotropic agent is guanidine HCl or urea.
22. The method of claim 19 , wherein the equilibration buffer is a phosphate buffer.