Immunoaffinity purification is a highly specific, reversible technique that has the potential to be used for the one-step isolation of an analyte from many complex matrices. Application of an immunoaffinity column to isolate and concentrate an analyte may decrease the amount of solvent used, decrease the number of purification steps, shorten analysis time, and simplify sample analysis relative to traditional clean-up techniques. In addition, immuno-affinity techniques may be easily integrated into high throughput systems. In this chapter, an immunoaffinity column generation method is outlined. Techniques and procedures for sample loading, washing, elution, and column storage will be described. In addition, methods used to determine column performance will be discussed. The information provided in this chapter will allow the inexperienced user to quickly become familiar with the generation, testing, and use of immunoaffinity columns.
Key Words: Immunoaffinity chromatography; IAC; isolation; purification; sample preparation; immunoaffinity columns.
From: Methods in Molecular Biology, vol. 295: Immunochemical Protocols, Third Edition. Edited by: R. Burns © Humana Press Inc., Totowa, NJ
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