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METHOD FOR PURIFICATION OF RECOMBINANT INTERFERON β-1b

机译:重组干扰素β-1b的纯化方法

摘要

FIELD: medicine.;SUBSTANCE: proposed method of purification of human recombinant interferon beta-1b (rIFN beta-1b) provides for optimisation of processes of purification of inclusion bodies, their dissolution and refolding of target protein Purification of human rIFN beta-1b is conducted with zwittehent 3-14 as a main detergent for purification and dissolving of inclusion body of Ecoli with the following refolding of rIFN beta-1b. After that prechromatographic processing of a sample is conducted and after that chromatographic processing of Ceramic S and of Source S30 and oxidation using a mixture of cysteamine/cystamine are performed. Then gel-filtration on Superdex 75 Prep grade, chromatographic processing on Source Q30, gel-filtration on Sephacril S200HR are made and finally the effectiveness of purification of the target protein by high performance liquid chromatography is monitored.;EFFECT: raise of the target protein yields and obtaining human rIFN beta-1b which meets the requirements of normative documents with high levels of specific antiviral activity.;5 tbl, 3 ex
机译:领域:医学;目的:拟议的纯化人重组干扰素β-1b(rIFN beta-1b)的方法可优化包涵体的纯化工艺,其溶解和靶蛋白的重折叠纯化人rIFN beta-1b的方法为用zwittehent 3-14作为主要去污剂进行纯化和溶解Ecoli包涵体,随后重新折叠rIFN beta-1b。之后,进行样品的预色谱处理,然后进行陶瓷S和源S30的色谱处理,并使用半胱胺/胱胺的混合物进行氧化。然后在Superdex 75 Prep级凝胶过滤,在Source Q30上进行色谱处理,在Sephacril S200HR上进行凝胶过滤,最后监测通过高效液相色谱纯化目标蛋白的有效性。产生并获得人rIFN beta-1b,其符合具有高水平抗病毒活性标准文件的要求。5tbl,3 ex

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