包含体中单链抗体3H11的复性条件对抗体活性的影响

THE AFFECTION OF RENATURATION CONDITIONS ON ACTIVITY OF SINGLE CHAIN ANTIBODY 3H11 FROM INCLUSION BODIES

研究包含体中单链抗体的最佳复性条件，并探讨复性后单链抗体的保存条件。方法：将含抗胃癌单链抗体3H11（ScFv3H11）的包含体纯化后在不同温度、分批加入、不同的变性液和复性液体积比的条件下进行复性，用ELISA法检测活性。结果：在10℃复性的抗体活性是4℃条件下的3倍；变性蛋白的分批加入及变性液和复性液的体积比为1：50时，均可分别获得相对最佳活性；复性液是复性抗体的理想保存条件。结论：此复性条件可以应用于其他原核表达的重组蛋白的复性。

Objective: The most recombinant antibodies were accumulated in inclusion bodies in E. coli. To get an optimum renaturation condition is essential for their activity. Metbods: ScFv 3H11 in purified inclusion bodies was refolded at different temperatures, different volume ratios of denature-renaturation solution and different addition times of denatured protein. Results: By measuring the activity of the refolded ScFv 3H11 with ELISA, it was found that the best renaturation temperature is 10℃, the best ratio is 1: 50. And frequent addition with small amount each time can improve the proportion of active protein relatively.Moreover, the stability of the ScFv 3H11 was also explored. Conclusion: This protocol may be used to rescue the activity of other recombinant proteins.