Unit

UNIT 6.12 Removal of Detergents from Proteins and Peptides in a Spin-Column Format

  1. Babu S. Antharavally

Published Online: 1 AUG 2012

DOI: 10.1002/0471140864.ps0612s69

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Antharavally, B. S. 2012. Removal of Detergents from Proteins and Peptides in a Spin-Column Format. Current Protocols in Protein Science. 69:6.12:6.12.1–6.12.7.

Author Information

  1. Thermo Fisher Scientific, Research & Development, Rockford, Illinois

Publication History

  1. Published Online: 1 AUG 2012
  2. Published Print: AUG 2012

Abstract

To enable downstream analysis, it is critical to remove unbound detergents from protein and peptide samples. This unit describes the use of a high-performance resin that offers exceptional detergent removal for proteins and peptides. The easy-to-use spin format significantly improves results over the standard drip column and batch methodologies, with >95% removal of 1% to 5% detergents, including SDS, sodium deoxycholate, CHAPS, Triton X-100, Triton X-114, NP-40, Brij-35, octyl glucoside, octyl thioglucoside, and lauryl maltoside, with high recovery of proteins and peptides. Detergent removal efficiency is evaluated using colorimetric methods and mass spectrometry (MS). BSA tryptic peptides have been successfully analyzed by liquid chromatography–tandem mass spectrometry (LC-MS/MS) and matrix-assisted laser desorption/ionization (MALDI)-MS for identification of protein, following detergent removal using the resin. Advantages of this method include speed (less than 15 min), efficient detergent removal, and high recovery of proteins and peptides. Curr. Protoc. Protein Sci. 69:6.12.1-6.12.7. © 2012 by John Wiley & Sons, Inc.

Keywords:

  • detergent removal;
  • spin-column format;
  • proteins and peptides;
  • downstream analysis;
  • mass spectrometry