Isolation and characterization of single-chain Fv genes encoding antibodies specific for p24 of HIV-1

By using phage display technology we generated recombinant mouse single chain antibody fragment directed against p24, the major capsid protein of human immunodeficiency virus type 1 (HIV-1). Using antibody variable-region (V) gene of B-cell derived from the spleen of an immunized mouse a library of single chain Fv fragments (scFv) was constructed. Following immunization of the mouse and preparation of scFv library, antibodies were selected by panning , using recombinant p24 protein which was expressed as a full length product in E.coli and purified by HPLC method. We isolated one single –chain antibody fragment which specifically recognizes p24 capsid protein of HIV-1. We showed the feasibility of the production of scFv in animal system (mice) by recombinant p24 cloning the repertoire of the heavy and light variable domains as a single chain Fv fragment (scFv), panning and selection, leading to the successful identification of a small antigen binder. It was observed that the antibody react specifically with p24 capsid protein which indicates that the Isolated mouse single-chain Fv fragment harbors the intact antigen binding site .Indeed specific antigen binder with good affinity was identified from this library, This antibody could be an ideal candidate for promising therapeutic applications and for HIV diagnosis. The isolated anti-p24 could be used in p24 antigen immunoassay system for detection of HIV-1 infection in an early phases . Indeed this antibody could be expressed as an intracellular antibody to aid treatment of HIV infection.

کلیدواژه ها:

HIV-1 ، p24 ، scFv ، Recombinant ، Antibody ، Phage display