抗HBV　Pre　S2单链噬菌体抗体基因在噬菌体载体中的克隆及初步表达

CLONING AND EXPRESSION OFA RECOMBINANTED ANTI-HBV PRE S2 SINGLE CHAIN PHAGED ANTIBODY GENE IN E.COLI

目的利用基因重组技术构建ＨＢＶ前Ｓ２单链抗体基因并使其在大肠杆菌中永生化和表达活性的抗ＨＢＶ前Ｓ２单链抗体（ＳｃＦｖ）。方法和结果利用基因重组技术，在构建抗ＨＢＶ前Ｓ２３Ｂ９ＳｃＦｖ基因的基础上，在此基因中引入限制性内切酶位点，克隆到噬菌体表达载体－ｐＣＡＮＴＡＢ５噬菌粒中，经亲和筛选得到２株阳性重组克隆，诱导表达产物在特异性阻断ＥＬＩＳＡ实验中具有一定阻断亲本３Ｂ９单抗与ＨＢＶ前Ｓ２抗原的结合能力，阻断率２２％－３３％。结论抗ＨＢＶ前Ｓ２ＳｃＦｖ在噬菌体中克隆和表达成功，为下一步继续表达单一的单链抗体并使其人源化打下了良好的物质基础。

Construction and immortalization of a single chain antibody(ScFv)gene in vitro.M6thods and Results The variable genes from 3B9 McAb were assembled into a single chain antibody(ScFv)gene by a linker and the ScFv gene was cloned into a expressive vector pCANTAB5 for expression of a recombinant phage ScFv, Two of the recombinant phagemids were characterized and the antigenic binding activities of the expressive recombinant phage ScFv were also detected by coated Pre S2 antigens and the blocking BLI SA(expressive phage ScFv samples were used to block the reaction of the parental 3B9 McAb). The blocking rates of the expressive phage ScFv to the 389 McAb was about 22%～33%.Conclusion The construction and expression of a phaged ScFv against HBV Pre S2 in E,coli were successful in this report.