The influence of water-miscible alcohols on the conformation of antibodies directed against a defined epitope.
Antiserum was raised against peptide P513 (CSQRSTNSAST) conjugated to diphtheria toxoid. P513 is a sequence derived from the Plasmodium falciparum merozoite surface antigen (MSA-2) containing the STNS epitope recognized by antibodies that inhibit parasite growth. Although antibodies directed against other STNS-containing peptides can bind to the native MSA-2 antigen, polyclonal anti-P513 serum failed to react in immunofluorescence assays with parasites or with MSA-2 by Western blotting. However, addition of methanol, ethanol or 1-propanol modified the specificity of the anti-P513 interaction, resulting in recognition of MSA-2. The effect was concentration dependent: at high alcohol concentrations, the specificity was further modified, and a previously undescribed rhoptry antigen was recognized. These results indicate that addition of alcohols to immunoassays may provide a way of modifying the specificity of antibodies in general, and the reactivity of anti-peptide serum with the parent protein in particular.
Lord
R
R
Queensland Institute of Medical Research, Australia.
Jones
G L
GL
Saul
A
A
D000433:1-Propanol / Q000737:chemistry; D000438:Alcohols / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000918:Antibody Specificity; D000953:Antigens, Protozoan / Q000737:chemistry / Q000276:immunology; D015153:Blotting, Western; D000939:Epitopes / Q000276:immunology*; D000432:Methanol / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D010963:Plasmodium falciparum / Q000276:immunology; D011487:Protein Conformation; D012995:Solubility; D014867:Water / Q000737:chemistry*
D000438:Alcohols; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D000939:Epitopes; D010455:Peptides; D014867:Water; D000433:1-Propanol; D000432:Methanol