摘要
目的快速制备大量具有生物活性的独特型纳米抗体F7。方法构建pET22b-F7原核表达载体,转化至大肠杆菌E.coliBL21(DE3)中进行诱导表达,对诱导温度、诱导剂浓度和诱导时间进行优化。结果独特型纳米抗体F7表达量有所增加但主要以包涵体形式存在。经过包涵体的溶解、复性获得了具有生物活性的抗AFB1独特型纳米抗体,ELISA证实复性后的蛋白依然存在对鼠源AFB1抗体的结合特性。结论为后续抗体的性质研究和改造奠定了基础。
Objective To rapidly produce a large amount of anti-idiotype nanobody against aflatoxin B1 (AFB1). Methods Prokaryotic expression vector pET22b-F7 were constructed, and then transformed into E.coli BL21(DE3) for expression of anti-idiotype nanobody against aflatoxin B1 (AFB1). The conditions such as temperature, concentration of inducer, and induction time were optimized. Results The expression quantity of anti-idiotype nanobody F7 increased, with the main form of inclusion body. Anti-idiotype nanobody against AFB1 with bioactivity were obtained after dissolution and renaturation. ELISA results indicated that the rena-tured protein had the properties of combination with AFB1 antibody. Conclusion It provides a reference for further study of properties and subsequent transformation.
出处
《食品安全质量检测学报》
CAS
2013年第4期1222-1227,共6页
Journal of Food Safety and Quality
关键词
黄曲霉毒素B1
独特型抗体
纳米抗体
原核表达
复性
aflatoxin B 1
anti-idiotype antibody
nanobody
prokaryotic expression
renaturation
