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Physicochemical Approach to the Purification of Human ₁-Fetoprotein from the Ascites Fluid of a Hepatoma-bearing Patient¹

Division of Clinical Immunology and Allergy [P. G., S. O. F., J. S.] and McGill University Medical Clinic, Montreal General Hospital Research Institute [P. G., A. L., H. C. G. W., S. O. F., J. K., J. S.], Montreal, Quebec H3G 1A4, Canada

A method for the purification of human ₁-fetoprotein from the ascites fluid of a hepatoma-bearing patient is described that is capable of yielding large quantities of pure ₁-fetoprotein within a relatively short period of time. The technique is based entirely on the physicochemical properties of the ₁-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 ₁-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 ₁-fetoprotein isolated. Amino acid analysis of the final product revealed a composition very similar to those reported for -fetoprotein preparations that have been previously isolated by the use of immunochemical technology.

¹ 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.

² Associate of the Medical Research Council of Canada.

³ Clinical Research Associate of the National Cancer Institute of Canada.

Received 12/20/76. Accepted 9/27/77.