Analytical high-performance affinity chromatography: evaluation by

Jul 15, 1986 - ... OF BIOPOLYMERS BY MEANS OF COLUMN LIQUID CHROMATOGRAPHY. K.K. Unger , R. Janzen , G. Jilge , K.D. Lork , B. Anspach. 1988 ...
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4148

Biochemistry 1986, 25, 4148-4155

Analytical High-Performance Affinity Chromatography: Evaluation by Studies of Neurophysin Self-Association and Neurophysin-Peptide Hormone Interaction Using Glass Matrices Harold E. Swaisgood*i’ and Irwin M. Chaiken* Molecular, Cellular, and Nutritional Endocrinology Branch, National Institute of Arthritis, Diabetes, and Digestive and Kidney Diseases, National Institutes of Health, Bethesda. Maryland 20205 Received August 27, 1985; Revised Manuscript Received March 6, 1986

I1 (BNP 11) was covalently immobilized on both nonporous and porous (200-nm pore diameter) glass beads and incorporated in a high-performance liquid chromatograph to evaluate analytical high-performance affinity chromatography as a microscale method for characterizing biomolecular interactions. By extension of the theoretical treatment of analytical affinity chromatography, both the self-association of neurophysin and its binding of the peptide hormone vasopressin were characterized by using a single chromatographic column containing immobilized neurophysin predominantly in the monomer form. Both [3H][Arg8]vasopressin (AVP) and ‘251-BNPI1 were rapidly eluted (