Power B E, Hudson P J
CSIRO Health Sciences and Nutrition, 343 Royal Parade, Parkville, Victoria 3052, Australia.
J Immunol Methods. 2000 Aug 28;242(1-2):193-204. doi: 10.1016/s0022-1759(00)00201-5.
Multivalent antibody fragments (scFv dimers, trimers and tetramers) provide high avidity and ideal pharmacokinetics for tumour targeting applications. This protocol describes our optimised protocol for high-level bacterial synthesis of soluble antibody scFv fragments, as either monomers or multimers, using the heat-inducible bacterial expression vector pPOW3. Our protocol is rapid, which minimizes protein degradation, and utilises inexpensive reagents for cost-effective product synthesis. The strong, temperature-regulated promoters in pPOW3 provide efficient production of either monomeric or multimeric single-chain antibody fragments as dictated by the gene construct design.
多价抗体片段（单链抗体二聚体、三聚体和四聚体）为肿瘤靶向应用提供了高亲和力和理想的药代动力学。本方案描述了我们优化的方案，用于使用热诱导细菌表达载体pPOW3在细菌中高水平合成可溶性抗体单链抗体片段，可作为单体或多聚体。我们的方案速度快，可将蛋白质降解降至最低，并使用廉价试剂进行具有成本效益的产品合成。pPOW3中强大的、温度调节型启动子可根据基因构建体设计高效生产单体或多聚体单链抗体片段。
