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目的：构建抗人精浆蛋白的单链抗体基因，可为进一步构建、表达抗人精浆蛋白单链抗体／羧肽酶Ａ融合蛋白，用于前列腺癌的抗体导向酶－前体药物疗法奠定基础。方法：利用加端ＰＣＲ技术，在已克隆的抗人精浆蛋白单克隆抗体ＶＨ和Ｖκ基因两端加上限制性酶切位点，再分别克隆入载体ｐＵＣ１９－ｌｉｎｋｅｒ中，构建ＶＨ－ｌｉｎｋｅｒ－Ｖκ形式的Ｅ４Ｂ７单链抗体基因。利用全自动荧光测序仪测定其序列，采用ＰＣ／Ｇｅｎｅ软件与已知的ＶＨ和Ｖκ基因进行核苷酸序列的同源性比较，并推导其编码的氨基酸序列。结果：Ｅ４Ｂ７单链抗体基因全长为７４１ｂｐ，为一开放读框，编码２４７个氨基酸，与已知的抗人精浆蛋白单克隆抗体ＶＨ和Ｖκ基因完全同源。ＶＨ和Ｖκ间有４５ｂｐ的ｌｉｎｋｅｒ序列，推导的氨基酸序列为（Ｇｌｙ４Ｓｅｒ）３，与设计的序列相符。结论：构建成功序列正确的抗人精浆蛋白单链抗体基因，为构建、表达抗人精浆蛋白／羧肽酶Ａ双功能抗体奠定了基础
OBJECTIVE: To construct a single-chain antibody gene for anti-human seminal plasma protein for further construction and expression of an anti-human seminal protein scFv / carboxypeptidase A fusion protein for the antibody directed enzyme-prodrug therapy of prostate cancer basis. Methods: The restriction endonuclease sites were added to the VH and Vκ genes of the cloned anti-human seminal plasma protein monoclonal antibody by means of additive-end PCR and cloned into vector pUC19-linker to construct VH-linker- VK form of the E4B7 single-chain antibody gene. The sequence was determined by using an automatic fluorescence sequencer. The homology of the nucleotide sequences was compared with that of the known VH and Vκ genes using PC / Gene software, and the deduced amino acid sequence was deduced. Results: The full length of the E4B7 single chain antibody gene was 741bp. It was an open reading frame and encoded a protein of 247 amino acids. It was completely homologous to the VH and Vκ genes of known anti - human seminal plasma monoclonal antibodies. The linker sequence of 45bp between VH and Vκ was deduced, and the deduced amino acid sequence was (Gly4Ser) 3, which was consistent with the designed sequence. CONCLUSION: The construction of a correct sequence of human anti-human seminal plasma protein chain antibody gene provides a basis for the construction and expression of anti-human seminoprotein / carboxypeptidase A bifunctional antibody