[Expression of human ScFv against human immunodeficiency virus type 1 gp120]

In order to generate human antibody against HIV-1, using synthetic peptide from HIV-1 gp120 v3 loop as antigen, we screened the recombinant phage antibodies by immunoaffinity and selected positive clones carrying anti-gp120 ScFv gene. The phage DNA was extracted and transformed to E. coli HB2151 that allowed to express soluble ScFv proteins by inducing of IPTG. The results of SDS-PAGE and Western blot showed that the molecular weight of the expressed product is about 28 KD and exhibited anti-c-myc activity. ELISA and Dot blot analysis displayed that the soluble ScFv specifically bound to gp120 and had a higher specificity and affinity. Competitive ELISA further demonstrated that the product was specific to gp120.