A method for the purification of human α1-fetoprotein from the ascites fluid of a hepatoma-bearing patient is described that is capable of yielding large quantities of pure α1-fetoprotein within a relatively short period of time. The technique is based entirely on the physicochemical properties of the α1-fetoprotein molecule and uses sequential purification steps: ion-exchange chromatography on DEAE-Sephadex A-50, molecular-sieve chromatography on Sephadex G-200, negative-affinity chromatography on Sepharose-Blue Dextran, positive-affinity chromatography on concanavalin A-Sepharose and, finally, molecular-sieve chromatography on Sephadex G-100. The efficiency of the entire procedure in its present form is 15% of the α1-fetoprotein activity of the starting preparation from ascites fluid. The purity of the final product was shown by polyacrylamide gel electrophoresis, radioimmunoelectrophoresis, and determinations of the NH2-terminal and COOH-terminal amino acid residues of the α1-fetoprotein isolated. Amino acid analysis of the final product revealed a composition very similar to those reported for α-fetoprotein preparations that have been previously isolated by the use of immunochemical technology.


This investigation was supported by grants from the Medical Research Council of Canada, Ottawa, Ontario, Canada, and the National Cancer Institute of Canada, Toronto, Ontario, Canada.

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