Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: December 18, 2002
Publication Date: January 1, 2005
Citation: Shelver, Weilin L. 2005. Generation and Testing of Immunoaffinity Columns IN: METHODS IN MOLECULAR BIOLOGY, Vol. 295: Immunochemical Protocols, Third Edition. Edited by R. Burns, Humana Press Inc., Totowa, NJ, pp: 269-280. Interpretive Summary: Immunoaffinity columns are extremely versatile and have been used for the isolation and concentration of a diverse number of analytes from a wide array of matrices. Because of its value as a research tool, immunoaffinity columns has found extensive use by the food safety community to isolate toxins or chemicals from food matrices. This chapter outline procedures for generation and testing of immunoaffinity columns.
Technical Abstract: 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, immunoaffinity techniques may be easily integrated into high throughput systems. In this chapter, an immunoaffinity column (IAC) 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 beginning user to quickly become familiar with the generation, testing, and use of immunoaffinity columns.