Isolation and Identification of α 1 -Antitrypsin as a Component of Normal and Malignant Human Breast and Other Tissues 1

Abstract

Summary

α₁-Antitrypsin has been detected using radial immunodiffusion in the 90,000g supernatant fraction of malignant and adjacent normal human breast, colon, and anal tissue, as well as malignant lung, stomach, and ileum. Immobilized chymotrypsin, bound to Sepharose or Affi-Gel 10 has been utilized to separate 11 peaks of antiproteolytic activity by affinity chromatography of normal and malignant human breast tissue extracts. Glycoproteins are associated with eight of the peaks. Peak 1 contains predominantly α₁-antitrypsin in addition to a minor component. The purification of the inhibitors, as judged by disc gel electrophoresis, is extensive. In some peaks, only one or two protein bands are observed, suggesting that affinity chromatography on Sepharose- or Affi-Gel 10-chymotrypsin might be used for the isolation of α₁-antitrypsin and other inhibitors in preparative amounts.

The authors would like to acknowledge the gift of our initial Sepharose-chymotrypsin from Dr. Melvin Keys, Owens Illinois Research Center, Toledo, Ohio. We are grateful to Dr. A. Bennett, Toledo Hospital, Toledo, OH, Drs. L. J. France and D. J. Hansen of Mercy Hospital, Toledo, OH, Dr. A. B. Batista, St. Lukes Hospital, Maumee, OH and Drs. J. R. Janney and B. Tancinco, Wood County Hospital, Bowling Green, OH for supplying the pathological tissue.