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Appl Environ Microbiol. 1974 July; 28(1): 133-137
Copyright © 1974 American Society for Microbiology. All Rights Reserved.

Comparative Study of ¹²⁵I- and [³H]Acetate-Labeled Antibodies in Detecting Iridescent Viruses

Department of Microbiology, University of Otago, Dunedin, New Zealand

ABSTRACT

Radioimmunoassays for detecting cell-associated or released virus are described using either ¹²⁵I- or [³H]acetate-labeled antibodies. In the first assay system, antigen-antibody complexes were separated from free antibody by centrifugation. Sensitivities of 0.1 µg of iridescent virus could be achieved with either ¹²⁵I- or [³H]acetate-labeled antibody. In the second assay, the antigen was fixed to cover-slip cell cultures, and then reacted with labeled antibody, unbound radioactivity being removed by repeated washing. Nonspecific binding with this method was 0.5 to 1% of the total radioactivity added and sensitivities of 0.1 or 10 µg were achieved with ¹²⁵I and [³H]acetate, respectively. Immunoglobulins were labeled at the rate of 1 in 300 for ¹²⁵I and 1 in 200 with [³H]acetate although there was a 400-fold greater isotopic abundance of ¹²⁵I relative to ³H. The possibility of preparing labeled protein of high specific activity using carrier-free [2-³H]iodoacetic acid is discussed.