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目的构建半合成抗体库，不经免疫制备抗体。方法通过ＰＣＲ法，从人胎肝ＤＮＡ扩增基因组ＶＨ段基因，从Ｆｄ基因制备含有不同长度随机化ＣＤＲ３的ＦＲ３－ＣＤＲ３－Ｊ－ＣＨ１片段，然后用重叠ＰＣＲ法构建Ｆｄ库，与人轻链基因库组合构建成半合成抗体库。结果通过各种组合多次建库，获得总容量为４×１０８的半合成抗体库，其Ｆｄ段和轻链基因的重组率均大于９０％，Ｆａｂ表达率为５０％。挑选２０个克隆测定ＣＤＲ３序列符合所设计的随机序列，用三种不同抗原进行筛选均得到了特异性噬菌体抗体。结论所构建的半合成噬菌体抗体库可以用于不经免疫制备抗体
Objective To construct a semi-synthetic antibody library and prepare antibodies without immunization. Methods The genomic VH gene was amplified from human fetal liver DNA by PCR and the FR3-CDR3-J-CH1 fragment containing randomized CDR3 of different length was prepared from Fd gene. Then the Fd library was constructed by overlap PCR, Gene bank combinations were constructed as semi-synthetic antibody repertoires. Results The semi-synthetic antibody library with a total capacity of 4 × 108 was obtained through multiple database construction. The recombination rate of Fd segment and light chain gene was over 90% and the expression rate of Fab was 50%. Twenty clones were selected to determine the CDR3 sequence in accordance with the designed random sequence. Specific phage antibodies were obtained by screening with three different antigens. Conclusion The constructed semi-synthetic phage antibody library can be used to prepare antibodies without immunization