Estradiol affinity chromatography: application to purification of murine alpha-fetoprotein

Journal of Chromatography
G J MizejewskiR Simon

Abstract

A one-step batch procedure is described for purification of murine alpha-fetoprotein (AFP) by estradiol affinity chromatography. Various ratios of carbodiimide (C), diaminononame (D) and estradiol hemisuccinate (E) were tested to determine optimal conditions for AFP purification. Although yields of AFP ranged from 15 to 44% depending on the reagent ratio employed, AFP isolates free of other protein contaminants were achieved at C:D:E ratios of 10:10:1 with a 29% yield. Both estrone and estradiol proved efficient as elution agents to free AFP bound to the estradiol-Sepharose beads, but higher yields were produced with estrone. After isolation the estrogen-eluted AFP preparations were analyzed by (1) estradiol-binding assays, (2) third-party radiocoprecipitation, (3) inhibition of radioimmunoassay for estrone and estradiol and (4) exchange of unlabeled for radiolabeled estradiol. These results indicated that the steroid remained attached to the eluted AFP molecule.

References

Jun 1, 1975·Journal of the National Cancer Institute·G J MizejewskiR P Allen
Apr 14, 1976·Biochimica Et Biophysica Acta·M S SoloffK Kithier
Mar 28, 1977·Biochimica Et Biophysica Acta·R P Allen, G J Mizejewski
Jul 5, 1974·Nature·G J Mizejewski, R P Allen
May 1, 1971·Proceedings of the National Academy of Sciences of the United States of America·J D Baxter, G M Tomkins
Sep 1, 1965·Immunochemistry·G ManciniJ F Heremans
Oct 1, 1968·Proceedings of the National Academy of Sciences of the United States of America·P CuatrecasasC B Anfinsen

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Citations

Dec 22, 1999·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·G M Butterstein, G J Mizejewski
Jan 1, 1983·Developmental and Comparative Immunology·G J Mizejewski, M Vonnegut
Dec 31, 2002·Expert Review of Anticancer Therapy·Gerald J Mizejewski
Jun 1, 2004·Experimental Biology and Medicine·Gerald J Mizejewski

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