Generation and identification of phage engineering antibodies library against hamster prion protein / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>Generation and Identification of Phage Engineering Antibodies Library against Hamster Prion Protein.</p><p><b>METHODS</b>Fab antibodies were identified and confirmed. BALB/c mice were immuned with PrP proteins. After the third immunization, the total RNA was extracted from the mice spleens. The genes of heavy Fd Fragments and light chain of antibodies amplified by a series of specific primers of human IgG Fab fragment were cloned into phagemid vector pComb3.</p><p><b>RESULTS</b>The combinatorial Fab library were constructed successfully and the cloning efficiencies both of light chain and Fd fragments were about near 10(6). The Fab library were panned by four cycles and screened with purified haPrP23-231 antigen on microtiter plates. 12 mAbs were isolated after four cycles of panning, five of which were sequenced and resulted sequence data were analyzed by alignment with GenBank immunoglobulin genes. Two strain of new heavy and light chain genes of Fab antibodies were identified and confirmed.</p><p><b>CONCLUSION</b>The research in this article will provide foundation for study of diagnosis and therapy of prion.</p>