RÉSUMÉ
<p><b>OBJECTIVE</b>To screen human monoclonal Fab fragments against HIV-1 gp120 peptide binding chemokine receptor.</p><p><b>METHODS</b>A synthesized polypeptide containing 23 amino acid residues of the gp120 antigen epitope binding chemokine receptor was coated as the solid-phase antigen. After biopanning from the HIV-1 phage Fab antibody library, the acquired positive clones were tested and sequenced.</p><p><b>RESULTS</b>One clone of human phage Fab monoclonal antibody against HIV-1 gp120 polypeptide was acquired. It has high affinity, specificity and inhibition rate and it belongs to IgG I subclass and kappa type. Its Vh H and V kappa were derived from Vh III and V kappa III.</p><p><b>CONCLUSIONS</b>The human phage Fab fragment against HIV-1 gp120 antigen site binding chemokine receptor was acquired.</p>
Sujet(s)
Humains , Séquence d'acides aminés , Anticorps monoclonaux , Génétique , Anticorps antiviraux , Génétique , Bactériophages , Génétique , Clonage moléculaire , Protéine d'enveloppe gp120 du VIH , Fragments Fab d'immunoglobuline , Génétique , Techniques in vitro , Données de séquences moléculaires , Banque de peptides , Récepteurs aux chimiokines , Métabolisme , Récepteur VIH , Allergie et immunologieRÉSUMÉ
<p><b>OBJECTIVE</b>To describe and evaluate a double-antigen sandwich ELISA for detecting human immunodeficiency virus type 1/2 (HIV-1/2) specific antibodies.</p><p><b>METHODS</b>The peptides gp41.1(sp1), gp41.2(sp2), gp120(sp3) and p24(sp4) of HIV-1 and gp36(sp5) of HIV-2 were artificially synthesized. Then sp1, sp3, sp4 and sp5 were used as coating antigens; sp1, sp2, sp4 and sp5 labeled with HRP were used as conjugates in this sandwich ELISA.</p><p><b>RESULTS</b>The specificity and sensitivity of the assay were both 100% in detecting anti-HIV of 40 control sera of the second generation panel, higher than indirect ELISA (specificity 90% and sensitivity, 65%, respectively). All of 210 sera from individuals with other diseases were negative for anti-HIV. The consistency rate was 100% when our sandwich ELISA and Abbott HIVAB were used to detect anti-HIV in 90 healthy blood donors and 88 HIV infected individuals.</p><p><b>CONCLUSIONS</b>The results showed that this sandwich ELISA for detection of anti-HIV is specific, sensitive and convenient, and it is suitable for screening blood donors and detecting HIV infection.</p>