Titration of penicilloyl antibody by enzyme-immunoassay: method and data analysis to assign titer.
Antisera to the hapten, penicilloyl, were titered in an indirect enzyme immunoassay with the marker enzyme, horseradish peroxidase. A multivalent solid-phase antigen for stable solid-phase immunocomplex adsorption and a 10 min peroxidase light-assay with photolabile ortho-phenylenediamine were features of the assay. The penicilloyl antisera titration curves formed the basis for an evaluation of 4 methods to assign titer. Titer was expressed as either the antiserum dilution-1 that produced an assigned absorbance, exhibited 50% maximal antigen-antibody binding, and equaled zero absorbance by extrapolation, or as absorbance at 10(-3) antiserum dilution. The accuracy for extrapolated titer was improved with 2 or more solid-phase antigen concentrations. Absorbance at 10(-3) antiserum dilution was most useful as a screen for relative titer and for specific titer from a standard curve generated with many test sera. The antiserum dilution may vary with a particular enzyme immunoassay.
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000918:Antibody Specificity; D044482:Benzeneacetamides; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006241:Haptens; D006735:Horseradish Peroxidase; D007124:Immunoenzyme Techniques; D010397:Penicillanic Acid / Q000276:immunology* / Q000031:analogs & derivatives; D010455:Peptides / Q000276:immunology*; D011107:Polylysine / Q000031:analogs & derivatives / Q000276:immunology*; D011817:Rabbits
D000906:Antibodies; D044482:Benzeneacetamides; D000939:Epitopes; D006241:Haptens; D010455:Peptides; D011107:Polylysine; C003058:penicilloyl polylysine; D010397:Penicillanic Acid; D006735:Horseradish Peroxidase