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应用氯化铯－超离心法从抗人纤维蛋白单克隆抗体ＳＺ一６３杂交瘤细胞抽取总ＲＮＡ，经Ｏｌｉｇｏ（ｄＴ）纤维素柱层析分离出ｍＲＮＡ。采用ＲＴ－ＰＣＲ技术，扩增出了ＳＺ－６３重链和轻链可变区基因。应用基因重组技术将ＳＺ－６３可变区基因片段与人免疫球蛋白γ１重链ＣＨ１和轻链恒区基因进行拼接，构建噬菌体ＳＺ－６３／ＨｕＦａｂ表达载体，并在大肠杆菌中表达。表达的嵌合抗体Ｆａｂ片段为可溶性，ＥＬＩＳＡ及Ｗｅｓｔｅｒｎｂｌｏｔ检测证实能特异地与人纤维蛋白呈结合反应，表达量约为２１０μｇ／Ｌ。
Total RNA was extracted from anti-human fibrin monoclonal antibody SZ-63 hybridoma cells by cesium chloride-ultracentrifugation and mRNA was isolated by Oligo (dT) cellulose column chromatography. The SZ-63 heavy chain and light chain variable region genes were amplified by RT-PCR. SZ-63 / HuFab expression vector was constructed by splicing SZ-63 variable region gene with human immunoglobulin γ1 heavy chain CH1 and light chain constant region gene, and expressed in E. coli. The expressed chimeric antibody Fab fragment was soluble, and ELISA and Western blot confirmed that it could specifically bind to human fibrin, and the expressed amount was about 210μg / L.