Abstract
A new procedure for the isolation of milligrams of AFP from a large volume of human cord serum is described. The purification steps include: 1. 1. batchwise chromatography on DEAE-cellulose, 2. 2. DEAE-cellulose chromatography, 3. 3. Sephadex G-200 column chromatography, 4. 4. Con A-Sepharose 4B column chromatography, 5. 5. Blue Sepharose column chromatography, 6. 6. immunoadsorbent column chromatography on columns filled with anti-cord serum-coupled Sepharose and anti-cord serum-coupled Sepharose but with a majority of anti-albumin being depleted. The procedure does not require an anti-AFP immunoadsorbent column. Usually more than 10 mg AFP may be obtained from 500 ml of cord serum. The purified AFP contains no measurable albumin and shows a single band after polyacrylamide gel electrophoresis. The purified AFP is suitable for raising anti-AFP and for the development of sensitive AFP radioimmunoassay.
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