METHOD FOR OBTAINING IMMUNOTROPIC SUBSTANCES OUT OF SWINE SKIN
FIELD: immunology, immunopharmacology, medicinal biotechnology, pharmaceutics. SUBSTANCE: method deals with reduction, homogenization of skin, centrifuging, thermodenaturation of supernatant, centrifuging of denaturated material, treatment of supernatant with cooled acetone, washing out of residue w...
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Zusammenfassung: | FIELD: immunology, immunopharmacology, medicinal biotechnology, pharmaceutics. SUBSTANCE: method deals with reduction, homogenization of skin, centrifuging, thermodenaturation of supernatant, centrifuging of denaturated material, treatment of supernatant with cooled acetone, washing out of residue with cooled acetone, drying, dissolving of residue and centrifuging, followed by chromatography at column with Sephadex G-50 and collection of fraction of target product at 1400-15000 Da molecular weight, desalinization and lyophilization, where centrifuging of dissolved residue is carried out at 8000-10000 g for 10-20 min at +4-6 C, one should add dropwise saturated solution of ammonium sulfate to supernatant at 2:1-3:1 ratio, centrifuge at 8000-10000 g for 40- 50 min at 4-6 C to achieve pH of obtained supernatant to be 3.5- 4.5 due to adding dropwise 8-10%-acetic acid solution, then dry ammonium sulfate should be added at 14-15 g/100 ml supernatant, centrifuged at 10000-12000 g for 20-30 min at 4-6 C, residue should be homogenized in 0.01-0.02 M tris HCl pH 7.5-8.5, then one should add dropwise saturated solution of ammonium sulfate to homogenate at 1:1-1.5:1 ratio, centrifuge at 16000-18000 g for 20-30 min at 4-6 C, residue is homogenized in 30-50 ml 0.01-0.02 M tris-HCl + 0.14-0.15 M NaCl pH 7.5-8.5, centrifuged at 16000-18000 g for 20-30 min at 4-6 C, and at chromatography upon column with Sephadex G-50 one should simultaneously collect fraction at molecular weight being above 15000 Da. The present innovation enables to increase activity of obtained immunotropic substances and obtain substances affecting both B- and T-systems of immunity. EFFECT: higher efficiency. 3 tbl |
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