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The Localization of Radioantibodies in Human Brain Tumors. III Radioiodination of Pre-purified Localizing Antibody^1,2

Durham, North Carolina

ABSTRACT

If an eventually isolated, specific glioma-localizing antibody is to serve as a carrier of therapeutic amounts of radioactivity, a method for high specific labeling of antibody must be obtained. In order to obtain high specific labeling of tissue-localizing antibodies with radioiodine, the radioiodination must be done after antibody purification. However, after localizing antibodies are purified by absorption on and elution from well-washed tissue sediments, the antibody eluates also contain eluted tissue proteins. These proteins compete for radioiodine and prevent high specific labeling of antibody. Therefore, their removal is necessary before effective radioiodination of pre-purified antibody can be accomplished. The experiments reported here demonstrate this problem and show how passage of eluates through columns of DEAE-A50-Sephadex helps to separate eluted antibody from eluted tissue protein. As a result, as much as a fifty-fold increase in specific labeling is obtained, although this varies from tissue to tissue. The rat kidney-antikidney and rat spleen-antispleen systems serve as models for the experiments and finally the human glioma-antiglioma system is also tested.

FOOTNOTES

¹ Divisions of Immunology and Neurosurgery, Departments of Microbiology, and Immunology and Surgery, Duke University Medical Center, Durham, North Carolina.

² Research was supported by A.E.C. Contract No. AT-(40-1)-3195, by USPHS Research Grant No. CA 07401-01 from the National Institutes of Health, and by grant No. T-344 from the American Cancer Society.