Rapid Preparation and Purification of Alkaline Phosphatase and 125I-Labeled Antibody by Minicolumn Gel Centrifugation Chromatography

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Gel centrifugation chromatography was used for the preparation, purification and concentration of enzyme and 125I-labeled antibody. Low-molecular-weight reactants were rapidly and efficiently removed from either 125I-labeled antibody or enzyme-antibody conjugates by centrifugation of the reaction mixture through a minicolumn of Sephadex. Further purification of both radiolabeled and enzyme labeled antibodies were possible by the application and elution of each labeled antibody through a minicolumn of Protein A Sepharose. Minicolumns were constructed from readily available inexpensive components and allowed the rapid preparation and purification of antibody of high specific enzyme, and radioactivity. No difference between the 125I, or enzyme labeled antibody detection limit could be detected by a direct solid-phase immunoassay.