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目的:从Tomlinson J噬菌体抗体库中筛选人源化抗Cry1Ac蛋白单链抗体(scFv)并进行鉴定。方法:以Cry1Ac蛋白为抗原包被固相,经4轮“吸附-洗脱-扩增”过程后,挑取单克隆,用ELISA法测定特异性结合活性,并对其进行基因序列测定。结果:经过4轮筛选,获得了2株能与Cry1Ac蛋白特异性结合的阳性克隆。结论:利用噬菌体抗体库技术可以不经过免疫制备出高特异性的人源化抗Cry1Ac蛋白scFv,为Cry1Ac毒素蛋白检测提供了条件。
OBJECTIVE: To screen and identify humanized anti-Cry1Ac protein single-chain antibody (scFv) from the Tomlinson J phage antibody library. Methods: The solid phase was coated with Cry1Ac as antigen. After 4 rounds of “adsorption-eluting-amplification” process, the monoclonal antibody was picked and the specific binding activity was determined by ELISA. The gene sequence was determined . Results: After 4 rounds of screening, two positive clones were obtained that could specifically bind with Cry1Ac protein. Conclusion: The highly specific humanized Cry1Ac protein scFv can be prepared without phage immunization using the phage antibody library technology, which provides the conditions for the detection of Cry1Ac toxin protein.