UNIT 29.12 Detergent Analysis in Protein Samples Using Mid-Infrared (MIR) Spectroscopy

Abstract

Quantitating relative levels of detergent present in protein preparations or samples derived from biological material, such as tissue or body fluids, is important because the presence of detergent may affect downstream analyses as well as protein structure/function. Especially because sample volumes, analysts’ available time, and other resources may be limited, a method that consumes little sample and that is rapid and simple is needed for detergent analysis. It would also be preferable to have a method that is generally applicable across many aliphatic chain-containing molecules with many different physical properties. In this unit, methods are described for analyzing detergents and proteins in detergent-protein mixtures using mid-infrared (MIR) spectroscopy. A protocol is also included for efficient removal of unbound detergents from a protein sample accompanied by MIR-based monitoring of both detergent and protein content. This rapid monitoring of sample preparation during the workflow enables users to make timely decisions about sample preparation strategies that maximize both analyte purity and yield. © 2015 by John Wiley & Sons, Inc.