从大容量抗体库中筛选抗TNF-α人单链抗体

Screening of human anti-TNF-α scFv from large phage library

目的:从天然的单链大容量噬菌体抗体库中筛选特异性的抗TNF-α的人单链抗体(scFv)。方法:以TNF-α为抗原,通过"吸附-洗脱-扩增"过程从大容量抗体库中筛选特异性抗体,对其抗原结合活性和序列进行分析鉴定。结果:经过4轮筛选,获得62个与TNF-α结合的阳性克隆,其中27个特异性结合的克隆,指纹分析有7种不同的可变区片段;其中5种获得可溶性表达;基因序列分析表明其轻重链分属VλⅠ和VHⅣ亚群。结论:利用单链大容量抗体库技术可以不经免疫制备获得特异性抗TNF-α的scFv,为今后的研究和应用提供依据。

AIM： To clone human anti-TNF-α scFv from large phage antibody library. METHODS： Panning of large phage library against TNF-α was conducted to select specific antibodies. The antigen binding activity and DNA sequence were determined and analyzed by ELISA, restriction enzyme map. RESULTS： After 4 rounds of panning, 62 positive clones were obtained and 27 clones had specific binding ability to TNF-α. DNA fingerprinting of the 27 clones showed 7 different stripes indicated 7 different positive clones. 5 of the 7 clones expressed soluble anti-TNF-α activity. DNA sequence analysis showed that the variable regions of these scFvs belonged to different subgroups. CONCLUSION： Human TNF-α scFv have been successfully obtained from large phage antibody library.