CAJ | 학술논문

研究了重组牛白介素2复性条件,如pH,蛋白浓度,DTT浓度,氧化剂(GSSG)和还原剂(GSH)的比例,Cu2+ 及L-Arg,低浓度的盐酸胍的添加,复性方法等.结果表明,复性液中添加20 mmol/mLDTT可以提高rBoIL-2的复性效率; 利用尿浓度梯度复性法3.5 mg/mL的rBoIL-2蛋白溶液复性率可达50%以上,测定活性效价达9.6×104 IU/mL.
연구료중조우백개소2복성조건,여pH,단백농도,DTT농도,양화제(GSSG)화환원제(GSH)적비례,Cu2+ 급L-Arg,저농도적염산고적첨가,복성방법등.결과표명,복성액중첨가20 mmol/mLDTT가이제고rBoIL-2적복성효솔; 이용뇨농도제도복성법3.5 mg/mL적rBoIL-2단백용액복성솔가체50%이상,측정활성효개체9.6×104 IU/mL.
Recombinant bovine interleukin-2 forms insoluble inclusion body when overexpressed in Escherichia coli, and must be denatured and renatured in vitro before acquiring biological activity. We systematically studied such conditions of renaturing rBoIL-2 as pH, protein concentration, DTT concentration, ratio of oxidizer (GSSG) to reducer (GSH), and additions of Cu2+ , L-Arginine and Guandine-HCl of low concentration, as well as renaturation methods. The results showed that adding 20 mmol/L DTT to denaturation solution could improve the efficiency of the subsequent rBoIL-2 renaturation, and the refolding rate of 3.5 mg/mL rBoIL-2 protein solution could reach 50% by means of urea gradient renaturation, in which the bioactivity of 9.6 × 104 IU/mL could be obtained.