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  • Title: Characterization of human prostatic acid phosphatase radioiodinated by four different procedures.
    Author: McCarthy RC, Markowitz H.
    Journal: Clin Chim Acta; 1983 Aug 31; 132(3):277-86. PubMed ID: 6616882.
    Abstract:
    Prostatic acid phosphatase was labeled by four different iodination procedures: chloramine-T, insoluble or soluble lactoperoxidase, and Bolton-Hunter. Enzyme labeled by the chloramine-T procedure had suboptimal precipitation characteristics in immunoassays, with 36 to 67% of the tracer precipitated in antibody excess. Analysis of the tracer on 7.5% sodium dodecyl sulfate polyacrylamide gel showed that as much as 30% of the radioactivity migrated away from the protein within 6 minutes after the electrophoretic run was begun. The nature of this fast migrating radioiodinated component was not determined. The failure to obtain a tracer labeled by the chloramine-T procedure with optimal immunoprecipitation characteristics led to investigation of alternate radioiodination procedures. Insoluble or soluble lactoperoxidase was less effective than chloramine-T in producing a tracer suitable for radioimmunoassay. In contrast, the Bolton-Hunter procedure produced a tracer that had optimal precipitation characteristics; 88 to 91% of the tracer was precipitated in antibody excess. The tracer was further purified by affinity chromatography to minimize the possibility of other protein contaminants compromising the specificity of a radioimmunoassay for the enzyme.
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