Abstract:

For further study of 2F5 and 4E10 antibody, 2F5-scFv and 4E10-scFv were expressed and purified from E.coli cells. The 2F5scFv and 4E10-scFv genes were amplified by overlapping PCR. Recombinant vectors pET28a/2F5-scFv and pET28a/4E1-scFv were constructed and transformed into E. coli BL21（DE3）and Rosetta-gami2（DE3）pLysS, and protein expression was induced with IPTG. 2F5-scFv and 4E10-scFv were expressed, with the molecular weight of 27 kD and 29 kD, respectively. The formed inclusion bodies in E. coli. Denatured single chain antibody proteins, which were purified by nickel chelating chromatography and refolded by dialysis, showed specific reactivates to both MPER antigens prepared from prokaryotic and eukaryotic expression systems.

Key words: HIV-1, gp41, MPER, 2F5, 4E10, Single-chain antibody