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The Journal of Immunology, 1968, 100: 187-193.
Copyright © 1968 by The American Association of Immunologists, Inc.

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A Cellulose Mercurial: Preparation, Properties and Use in Antibody Purification1

John R. Shainoff

Research Division, The Cleveland Clinic Foundation, Cleveland, Ohio

Abstract

A cellulose mercurial has been developed and applied as an aid to the antibody isolation method that is based on use of thiolated antigens as retrievable harvesting agents. The mercurial was prepared by etherifying cellulose with 1-allyloxy-2,3-epoxypropane, and subsequently mercurating allyl groups with mercuric acetate. The mercurial had properties of an ion exchange resin with covatently bound, monofunctional HgII as exchanging group. In 0.05 M chloride at pH near 2.4, the resin seemed to bind protein only by forming mercaptides and, then, only with particular thiol groups. Native thiol groups of serum albumin or ovalbumin did not react, but groups added de novo by thiolating protein with N-acetyl homocysteine thiolactone did. In enabling separation of thiolated from non-thiolated proteins in presence of chloride, the resin provided an important advantage over a previously described mercurial. The resin was applied to isolation of anti-bovine {gamma}-globulin antibody. In this application, thiolated bovine {gamma}-globulin was used to precipitate the antibody. The precipitate was dissolved, and the thiolated antigen subsequently removed with the resin, leaving purified antibody in solution.

Footnotes

This investigation was supported in part by Grant HE-6835-06 of the National Institutes of Health. Presented in part at the Ninth Annual Meeting of the Biophysical Society in San Francisco, February, 1965.







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