Claims
- 1. In a method for the extraction and purification of biologically active lymphokines from large scale culture supernatants comprising the steps of effecting the growth of lymphoblastoid cells from human lymphoblastoid cultured cell lines, and thereafter harvesting, concentrating and clarifying the resulting supernatant culture fluids, the improvement which comprises extracting said supernatant culture fluids with a solvent selected from the group consisting of trichloroacetic acid, guanidine hydrochloride, sodium dodecyl sulfate and perchloric acid and removing said solvent to produce a lymphokine fraction which exhibits the capability of inducing cell-mediated immunity reactions and tumor regression in mammals.
- 2. The method as set forth in claim 1 wherein said lymphokine fraction is further purified by treatment with a dissociating solvent selected from the group consisting of guanidine hydrochloride and sodium dodecyl sulfate.
- 3. The method as set forth in claim 1 wherein said solvent is removed by gel filtration.
- 4. The method as set forth in claim 1 wherein said lymphokine fraction is subjected to further purification by a procedure selected from the group consisting of gel filtration, sodium dodecyl sulfate gel electrophoresis and high pressure liquid chromatography.
- 5. The method as set forth in claim 1 wherein the concentration of trichloroacetic acid is approximately 10% w/v.
- 6. The method as set forth in claim 2 wherein said dissociating solvent is 6 molar guanidine hydrochloride.
- 7. The method as set forth in claim 2 wherein said dissociating solvent is a 1% sodium dodecyl sulfate solution.
- 8. The method of producing lymphokine fraction which exhibits the capability of inducing cell-mediated immunity reactions and tumor regression in mammals by the extraction and purification of biologically active lymphokines from large scale culture supernatants comprising the steps of effecting the growth of lymphoblastoid cells from human lymphoblastoid cultured cell lines, harvesting, concentrating and clarifying the resulting supernatant culture fluids, extracting said supernatant fluids with a solvent selected from the group consisting of trichloroacetic acid, guanidine hydrochloride, sodium dodecyl sulfate and perchloric acid, and removing said solvent.
- 9. The method as set forth in claim 8 wherein said lymphokine fraction is further purified by treatment with a dissociating solvent selected from the group consisting of guanidine hydrochloride and sodium dodecyl sulfate.
- 10. The method as set forth in claim 8 wherein said solvent is removed by gel filtration.
- 11. The method as set forth in claim 8 wherein said lymphokine fraction is subjected to further purification by a procedure selected from the group consisting of gel filtration, sodium dodecyl sulfate gel electrophoresis and high pressure liquid chromatography.
- 12. The method as set forth in claim 8 wherein the concentration of trichloroacetic acid is approximately 10% w/v.
- 13. The method as set forth in claim 9 wherein said dissociating solvent is 6 molar guanidine hydrochloride.
- 14. The method as set forth in claim 9 wherein said dissociating solvent is a 1% sodium dodecyl sulfate solution.
- 15. The method of inducing cell-mediated immunity reactions and tumor regression in mammals comprising administering thereto a lymphokine fraction produced by effecting the growth of lymphoblastoid cells from human lymphoblastoid cultured cell lines, harvesting, concentrating and clarifying the resulting supernatant culture fluids, extracting aid supernatant culture fluids with a solvent selected from the group consisting of trichloroacetic acid, guanidine hydrochloride, sodium dodecyl sulfate and perchloric acid, and removing said solvent.
- 16. The method as set forth in claim 15 wherein said lymphokine fraction has been further purified by treatment with a dissociating solvent selected from the group consisting of guanidine hydrochloride and sodium dodecyl sulfate.
- 17. The method as set forth in claim 15 wherein said solvent is removed by gel filtration.
- 18. The method as set forth in claim 15 wherein said lymphokine fraction has been subjected to further purification by a procedure selected from the group consisting of gel filtration, sodium dodecyl sulfate gel electrophoresis and high pressure liquid chromatography.
- 19. The method as set forth in claim 15 wherein the concentration of trichloroacetic acid is approximately 10% w/v.
- 20. The method as set forth in claim 16 wherein said dissociating solvent is 6 molar guanidine hydrochloride.
- 21. The method as set forth in claim 16 wherein said dissociating solvent is a 1% sodium dodecyl sulfate solution.
- 22. The method as set forth in claim 15 wherein said lymphokine fraction is administered intravenously.
- 23. The method as set forth in claim 15 wherein said lymphokine fraction is administered intradermally.
- 24. The method as set forth in claim 15 wherein said lymphokine fraction is administered subcutaneously.
- 25. The method as set forth in claim 15 wherein said lymphokine fraction is administered as an adjuvant to chemotherapy.
Government Interests
The invention described herein was made in the course of work under a grant or award from the Department of Health, Education and Welfare.
Non-Patent Literature Citations (1)
Entry |
"Nature", vol. 224, Oct. 4, 1969, pp. 38-42, Article by Dumonde et al. |