Preparation and characterization of four novel monoclonal antibodies specific to N51(L6)C46 polypeptide simulating fusogenic core structure of GP41 subunit of HIV-1

We have generated monoclonal antibodies (MAbs) against the N51(L6)C46 polypeptide, which could stimulate the fusogenic core structure of the gp41 subunit of the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA) using either N51(L6)C46 or N36(L6)C34 polypeptide. The MAb against N36(L6)C34, NC-1, was used as positive control, and N36 C34 peptides as negative controls. As a result, we generated four MAbs, three (1A2, 1A4, 1B9) against the polypeptide N51(L6)C46 monomer and one against the trimer, named 3A1. The four anti- N51(L6)C46 MAbs could bind N51(L6)C46 and N36(L6)C34 but not N36 or C34. The results of epitope analysis with competitive ELISA showed that the MAbs could recognize the similar epitopes, whereas the difference lie in that 3A1 could recognize the epitopes similar to NC-1s, whereas NC-1 could not recognize the epitopes similar to 3A1s. Taken together, these MAbs would be effective tools in selecting anti-HIV polypeptide and analyzing the characteristics of gp41 epitopes.