Comparison of specific radioactivities of human alpha-lactalbumin iodinated by three different methods.
Radioiodination provides an extremely sensitive method for the detection of low levels of proteins. In the development of a sensitive radioimmunoassay for human alpha-lactalbumin (alpha-LA), the protein was labeled to high specific activity (approaching 2000 Ci/mmol) with lactoperoxidase, chloramine-T, and Iodogen. Despite high specific activities of the labeled protein by each method, there was a considerable difference in their binding affinity with monoclonal anti-human alpha-LA antibodies due to varying degrees of protein damage. Iodination of human alpha-LA with Iodogen resulted in labels of the highest specific activity and immunoreactivity with the monoclonal antibodies used.
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Department of Pathology & Immunology, Monash Medical School, Prahran, Victoria, Australia.
D000911:Antibodies, Monoclonal; D002700:Chloramines / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D007768:Lactalbumin / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D007784:Lactoperoxidase / Q000378:metabolism; D011233:Precipitin Tests; D011863:Radioimmunoassay / Q000379:methods*; D014105:Tosyl Compounds; D014508:Urea / Q000031:analogs & derivatives / Q000378:metabolism
D000911:Antibodies, Monoclonal; D002700:Chloramines; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D014105:Tosyl Compounds; C016300:chloramine-T; C012385:1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenylglycoluril; D014508:Urea; D007768:Lactalbumin; D007784:Lactoperoxidase