Adsorption resin AAS was used for the separation and purification of milupeinan (M-13) from its mother liquor. The good adsorption property of AAS adsorption resin was found for milupeinan (M-13) compared with the Dia...Adsorption resin AAS was used for the separation and purification of milupeinan (M-13) from its mother liquor. The good adsorption property of AAS adsorption resin was found for milupeinan (M-13) compared with the Diaion CHP 20p. When acetone and water (2:1 in volume) were used as desorption reagents, it was found that 3 times of bed volume of the desorption reagents would make the AAS resin which was saturately adsorpted M-13 desorption completely. Also a macroporous anion exchange resin (Poly-4-vinylpyridine type) D-280 was used for the decolourization from M-13 solution. Good results was given when the sv=1~2.展开更多
Alpha-interferon 2b (IFN 2b) was produced both in soluble and insoluble forms from recombinant E. coli. The dissolution of the expressed IFN 2b in inclusion body was carried out and it was found that the optimal condi...Alpha-interferon 2b (IFN 2b) was produced both in soluble and insoluble forms from recombinant E. coli. The dissolution of the expressed IFN 2b in inclusion body was carried out and it was found that the optimal condition to dissolve the expressed protein was 7 mol稬1guanidinium salt solution at pH 3.0. The resultant solution was diluted 20 times using pH 6.0 buffer to refold the protein correctly. The cation exchange column was employed to purify both refolded and soluble IFN 2b. For soluble IFN sample, high IFN 2b recovery yield (92.1%) with 91.7% purity was obtained in the eluate. However, for refolded IFN sample, only 72.7% of IFN 2b was recovered with relatively low purity (56.8%) by cation exchange chromatography. Although the expression level of insoluble IFN was higher than that of co-expressed soluble IFN in this recombinant E. coli cells, the productivity of bioactive IFN 2b was higher with soluble expressed IFN after primary purification process. Soluble expression of foreign proteins in recombinant bacteria might be an alternative strategy for efficient production of heterogeneous pro-teins due to high bioactivity and simple downstream protein purification process.展开更多
文摘Adsorption resin AAS was used for the separation and purification of milupeinan (M-13) from its mother liquor. The good adsorption property of AAS adsorption resin was found for milupeinan (M-13) compared with the Diaion CHP 20p. When acetone and water (2:1 in volume) were used as desorption reagents, it was found that 3 times of bed volume of the desorption reagents would make the AAS resin which was saturately adsorpted M-13 desorption completely. Also a macroporous anion exchange resin (Poly-4-vinylpyridine type) D-280 was used for the decolourization from M-13 solution. Good results was given when the sv=1~2.
文摘Alpha-interferon 2b (IFN 2b) was produced both in soluble and insoluble forms from recombinant E. coli. The dissolution of the expressed IFN 2b in inclusion body was carried out and it was found that the optimal condition to dissolve the expressed protein was 7 mol稬1guanidinium salt solution at pH 3.0. The resultant solution was diluted 20 times using pH 6.0 buffer to refold the protein correctly. The cation exchange column was employed to purify both refolded and soluble IFN 2b. For soluble IFN sample, high IFN 2b recovery yield (92.1%) with 91.7% purity was obtained in the eluate. However, for refolded IFN sample, only 72.7% of IFN 2b was recovered with relatively low purity (56.8%) by cation exchange chromatography. Although the expression level of insoluble IFN was higher than that of co-expressed soluble IFN in this recombinant E. coli cells, the productivity of bioactive IFN 2b was higher with soluble expressed IFN after primary purification process. Soluble expression of foreign proteins in recombinant bacteria might be an alternative strategy for efficient production of heterogeneous pro-teins due to high bioactivity and simple downstream protein purification process.
