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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 screen and identify human anti-keratin antibodies from semi-synthetic phage antibody library. Methods: The specific anti-keratin antibody was screened from the semi-synthetic phage antibody library by using the epidermal keratin as an antigen, and its antigen binding activity and sequence were identified and identified by adsorption-elution-amplification. Results: After 4 rounds of screening, 20 positive clones that could bind to keratin were obtained, of which 18 could produce specific anti-keratin antibodies. DNA fingerprinting analysis, the clones were determined to contain four different Fab segment genes. One clone after excision of gene Ⅲ expressed soluble Fab which could specifically bind with keratin. Sequence analysis showed that V and VH belonged to human V1 subgroup and VH1 subgroup respectively. The VHCDR3 sequence was consistent with the primers used in the construction of semi-synthetic antibody library design. CONCLUSIONS: Highly specific human anti-keratin antibodies can be prepared without phage using phage antibody library technology.