Immobilized Biochemicals and Affinity Chromatography

This volume contains most of the papers presented at the Sym posium on Affinity Chromatography and Immobilized Biochemicals, which was held on November 7-9, 1973 in Charleston, South Carolina in conjunction with the Southeastern Regional American Chemical Society meeting. The topics of the symposium represent two new biochemical frontiers which have emerged in recent years through the ingenious development and application of solid phase biochemi cal technologies. Affinity chromatography involves the use of selected ligands, covalently bound to a solid support such as cel lulose, glass, Sepharose, or polyacrylamide and exploits the princi ple of biochemical recognition between the ligand in the solid phase and a selected macromolecule to facilitate the rapid and often quantitative purification of enzymes, antibodies, antigens, hormones, other proteins, etc. The area of immobilized biochemicals includes the use of coenzymes, oligo- and po1ynuc1eotides, enzymes, and multistep enzyme systems which are immobilized or entrapped in the solid phase. The goals of the symposium were the revi~w of the status of affinity chromatography and immobilized biochemicals, the presentation of new data and ideas in both areas, and the establish ment of a dialogue between research workers in these two evolving disciplines which are so closely interrelated. The papers published in this volume provide the reader with reviews of several topics inherent in the solid phase biochemistry together with a series of timely manuscripts concerning new techniques and applications both in the use of affinity chromatography and in the investigation of immobilized biochemicals.

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One: Affinity Chromatography.- Affinity Chromatography--Old Problems and New Approaches.- Affinity Chromatography. New Approaches for the Preparation of Spacer Containing Derivatives and for Specific Isolation of Peptides.- Quantitative Parameters in Affinity Chromatography.- Non-Specific Binding of Proteins by Substituted Agaroses.- A Solid Phase Radioimmune Assay for Ornithine Transcarbamylase.- Purification of Acetylcholinesterase by Covalent Affinity Chromatography.- Cooperative Effects of AMP; ATP, and Fructose 1,6-Diphosphate on the Specific Elution of Fructose 1,6-Diphosphatase from Cellulose Phosphate.- An Analysis of Affinity Chromatography Using Immobilised Alkyl Nucleotides.- Affinity Chromatography of Kinases and Dehydrogenases on Sephadex and Sepharose Dye Derivatives.- Affinity Chromatography of Thymidylate Synthetases Using 5-Fluoro-2?-Deoxyuridine 5?-Phosphate Derivatives of Sepharose.- The Biosynthesis of Riboflavin: Affinity Chromatography Purification of GTP-Ring-Opening Enzyme.- Purification of Tyrosine-Sensitive 3-Deoxy-D-Arabino-heptulosonate-7-Phosphate and Tyrosyl-tRNA Synthetase on Agarose Carrying Carboxyl-Linked Tyrosine.- Structural Requirement of Ligands for Affinity Chromatography Absorbents: Purification of Aldehyde and Xanthine Oxidases.- Two: Immobilized Biochemicals.- Immobilized Polynucleotides and Nucleic Acids.- Immobilized Cofactors and Multi-Step Enzyme-Systems.- Preparation, Characterization, and Applications of Enzymes Immobilized on Inorganic Supports.- Lactase Immobilized on Stainless Steel and Other Dense Metal and Metal Oxide Supports.- The Use of Membrane-Bound Enzymes in an Immobilized Enzyme Reactor.- The Optimization of Porous Materials for Immobilized Enzyme Systems.- Water Encapsulated Enzymes in an Oil-ContinuousReactor: Kinetics and Reactivity.- Analysis of Reactions Catalyzed by Polysaccharide-Enzyme Derivatives in Packed Beds.- The Preparation of Microenvironments for Bound Enzymes by Solid Phase Peptide Synthesis.- Optimization of Activities of Immobilized Lysozyme, ?-Chymotrypsin, and Lipase.- Chemical Modification of Mushroom Tyrosinase for Stabilization to Reaction Inactivation.- Chain Refolding and Subunit Interactions in Enzyme Molecules Covalently Bound to a Solid Matrix.- Immobilization of Lipase to Cyanogen Bromide Activated Polysaccharide Carriers.- Use of Immobilized Enzymes for Synthetic Purposes.