UV-treated polystyrene microtitre plates for use in an ELISA to measure antibodies against synthetic peptides.
Detection of peptide-specific antibodies by the conventional ELISA technique is sometimes hampered by the difficulties encountered in immobilizing stretches of amino acids on the solid support. To improve the attachment of synthetic peptides to the solid phase, we have developed a sensitive and rapid immunoassay based on the irradiation of polystyrene plates with UV light prior to coating the target peptide. This pretreatment increases the specific signal in a dose-dependent manner without augmenting the background or altering the specificity of the assay. This simple method was shown to be suitable for the quantitation of murine monoclonal antibodies as well as human and rabbit polyclonal antibodies. It should be applicable to a variety of synthetic peptides and polystyrene ELISA plates. Using this technique, we were able to localize the antigenic motifs recognized by neutralizing monoclonal antibodies generated against the envelope protein gp120 of the human immunodeficiency virus.
Boudet
F
F
Institut Pasteur, Unité d'Immunogénétique Cellulaire, Paris, France.
Thèze
J
J
Zouali
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology; D005260:Female; D015483:HIV Antibodies / Q000032:analysis*; D015699:HIV Envelope Protein gp120 / Q000032:analysis*; D015497:HIV-1 / Q000276:immunology; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis*; D011137:Polystyrenes / Q000528:radiation effects; D011485:Protein Binding / Q000528:radiation effects; D011817:Rabbits; D012680:Sensitivity and Specificity; D014466:Ultraviolet Rays
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D010455:Peptides; D011137:Polystyrenes