摘要
目的：从半合成噬菌体抗体库中筛选人源性抗角蛋白抗体并进行鉴定。方法：以表皮角蛋白为抗原，通过吸附－洗脱－扩增过程从半合成噬菌体抗体库中筛选特异性抗角蛋白抗体，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２０个能与角蛋白结合的阳性克隆，其中可产生特异性抗角蛋白抗体的克隆１８个。经ＤＮＡ指纹分析，判断所获克隆分别包含４个不同的Ｆａｂ段基因。经切除基因Ⅲ后１个克隆表达出可与角蛋白特异性结合的可溶性Ｆａｂ，序列分析表明其Ｖ和ＶＨ分别属于人Ｖ１亚群和ＶＨ１亚群，ＶＨＣＤＲ３序列符合半合成抗体库构建时的引物设计。结论：利用噬菌体抗体库技术可以不经免疫制备出高特异性的人源性抗角蛋白抗体。
Objective: To clone human anti-keratin antibodies from semisynthetic phage antibody library. Methods: Panning of semisynthetic phage andtibody library against human epidermal keratin was conducted to elect specific antibodies. The antigen binding activity and DNAsequence were determined and analyzed. Results: After 4 rounds of panning,20 positive clones were obtained and 18 clones were found to beable to bind keratin antigen specifically. DNA fingerprinting of the 18 clones showed 4 different digestion patterns representing 4 different positive clones . One of them could express soluble anti-keratin Fab antibody after removing the gene III fragment . DNA sequence analyses showedthe VK and VH belonged to human VK1 subgroup and VH1 subgroup respectively. Conclusion: It is feasible to isolate human anti-keratin andtibodies by-passing immunization using phage antibody library.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2000年第1期5-7,共3页
Chinese Journal of Immunology
基金
国家自然科学基金!39670679
军队医药卫生重点课题!96Z0004
