论文部分内容阅读
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。
The optimal refolding conditions of the single-chain antibody in the inclusion body were studied, and the preservation conditions of the single-chain antibody after refolding were explored. Methods: The inclusions containing anti-gastric cancer single chain antibody 3H11 (ScFv3H11) were purified and renaturated at different temperatures, in batches, volume ratio of denatured solution to renaturation solution, and the activity was determined by ELISA. Results: The activity of renaturation at 10 ℃ was three times of that at 4 ℃. The best relative activity was obtained when the denatured protein was added in batches and the volume ratio of denatured solution to renaturation solution was 1:50. Refolding fluid is the ideal storage condition for refolding antibodies. Conclusion: The refolding conditions can be applied to the renaturation of recombinant proteins expressed in other prokaryotes.