By Pascal Bailon
Information strong affinity chromatography tools, starting from conventional affinity purification corresponding to immunoaffinity chromatography, to using the most recent phage-display expertise within the discovery of affinity ligands and medicine. additionally incorporated are separations of small molecules reminiscent of haptens, protein ligands, and supramolecular constructions. every one bankruptcy is dedicated to a selected approach and contains an advent, an evidence of rules, an in depth fabrics checklist, and directions. useful notes recommend replacement tactics and describe how one can triumph over difficulties. The editor works for an enormous pharmaceutical corporation.
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Additional info for Affinity Chromatography Methods and Protocols
6. 7. 8. 9. 10. 51 Equilibrate the Protein A column with 5 column volumes (cv) of buffer A. Load supernatant onto Protein A column at 5 mL/min. Load up to 20 mg antibody/mL gel. Wash with 10 cv of buffer B. Elute antibody with 5 cv of buffer C. Re-equilibrate column with 2 cv of buffer A. Wash with 5 cv of buffer E followed by 10 cv of buffer A. The column is now ready for the next chromatography run. 3. Notes This general protocol works well with high-affinity antibodies and is independent of the particular Protein A gel used.
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