Zhang Jian-Lin, Wang Jian-Hua, Zhong Ru-Gang, Niu Bo
Department of Biochemisty and Molecular Biology, Shanxi Medical University, Taiyuan 030001, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009 Feb;23(1):50-2.
To explore the conditions for high expression of anti-HBsAg scFv A-15 in E. coli, increase the production of the scFv in the culture medium.
By changing induction occasion, concentration of inductor IPTG and induction time, influence of various conditions on expression of anti-HBsAg scFv A-15 was analyzed through ELISA. In addition, the effects of sucrose, glycine and Triton X-100 at different concentrations on the scFv excretion into culture medium was evaluation.
The optimal expression conditions were as follows: the induction was started after culturing for 4 h, the concentration of IPTG was 0.5 mmol/L, and the induction lasted for 8 h. The scFv affinity in culture medium with 0.3 mol/L sucrose, 2% glycine, 1% Triton X-100, 16.78-fold higher, respectively than that without the three chemicals. The final yield of anti-HBsAg scFv A-15 was estimated to be 7.4 mg/L.
The conditions for production of anti-HBsAg scFv A-15 were optimized, which provides a practical method for more efficient production of the scFv in E. coli for further studying structure and function.
探索抗HBsAg单链抗体A-15在大肠杆菌中高效表达的条件，提高其在培养基中的产量。
通过改变诱导时机、诱导剂IPTG浓度和诱导时间，采用ELISA分析各种条件对抗HBsAg单链抗体A-15表达的影响。此外，评估不同浓度的蔗糖、甘氨酸和Triton X-100对单链抗体分泌到培养基中的作用。
最佳表达条件如下：培养4 h后开始诱导，IPTG浓度为0.5 mmol/L，诱导持续8 h。在含有0.3 mol/L蔗糖、2%甘氨酸、1% Triton X-100的培养基中，单链抗体的亲和力分别比不含这三种化学物质时高16.78倍。抗HBsAg单链抗体A-15的最终产量估计为7.4 mg/L。
优化了抗HBsAg单链抗体A-15的生产条件，为在大肠杆菌中更高效地生产单链抗体以进一步研究其结构和功能提供了一种实用方法。
