摘要
重组人粒 -单系集落刺激因子 (rh GM- CSF)在大肠杆菌表达后积累在细胞中形成包涵体 ,在变性的包涵体的抽提液复性过程中 ,目的产物可能聚积而使活性蛋白的得率降低。本研究对聚乙二醇 (PEG)增加复性的能力进行试验。将 rh GM- CSF在 6 mol/L尿素中变性 ,迅速稀释到不同浓度尿素和 PEG的溶液中并保持蛋白浓度为 0 .2 mg/ml,在尿素浓度为 2 .0 mol/L,PEG4 0 0 0对rh GM- CSF的摩尔比为 2 0∶ 1时活性蛋白从 4 0 %增加到 55% ,rh GM- CSF的比活从 5.1× 10 6u/mg增加到 1.0× 10 7u/mg,总蛋白得率从 50 %增加到 10 0 %。表明 PEG能增加变性蛋白的复性效率。
During refolding process,the protein could aggregate and the recovery of active product was reduced This paper tested the ability of polyethylene glycol(PEG) to enhance refolding rhGM CSF was unfolded in 6mol/L urea and rapidly diluted to 2 or 4mol/L urea and 0 2mg/ml protein 2mol/L urea and a PEG4000 to rhGM CSF molar ratio of 20 to 1 increased the yield of active protein from 40% to 55%,specific activity of rhGM CSF from 5 1×10 6u/mg to 1 0×10 7u/mg,total protein recovery from 50% to 100% It proved that PEG has significant ability for enhancing the refolding of unfolded proteins
出处
《药物生物技术》
CAS
CSCD
1997年第1期5-8,共4页
Pharmaceutical Biotechnology
关键词
集落刺激因子
聚乙二醇
复性
Recombinant human granulocyte macrophage colony stimulating factor,Polyethylene glycol,Unfolding,Refolding
