title: Efficient removal of detergents from proteins and peptides in a spin column format.
abstract: Detergents are commonly used in protein-chemistry protocols and may be necessary for protein extraction, solubilization, and denaturation; however, their presence interferes with many downstream analysis techniques, including mass spectrometry (MS). To enable downstream analysis, it is critical to remove unbound detergents from protein and peptide samples. In this study, we describe a high-performance resin that offers exceptional detergent removal for proteins and peptides. When used in a spin column format, this resin dramatically improves protein and peptide MS results by more than 95% removal of 1-5% detergents, including sodium dodecyl sulfate (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. Postcolumn liquid chromatography-tandem MS (LC-MS/MS) analysis of trypsin digests of bovine serum albumin (BSA) and HeLa cell lysate revealed excellent sequence coverage, indicating successful removal of detergent from the peptides. Matrix-assisted laser desorption/ionization (MALDI)-MS analysis of unprocessed and processed samples further confirmed efficient removal of detergents. The advantages of this method include speed (<15min), efficient detergent removal, and high recovery of proteins and peptides.
authors.affiliation: Thermo Scientific Pierce Protein Research, Thermo Fisher Scientific, Rockford, IL 61105, USA. babu.antharavally@thermofisher.com
authors.forename: Babu S
authors.initials: BS
authors.lastname: Antharavally
authors.forename: Krishna A
authors.initials: KA
authors.lastname: Mallia
authors.forename: Michael M
authors.initials: MM
authors.lastname: Rosenblatt
authors.forename: Ashok M
authors.initials: AM
authors.lastname: Salunkhe
authors.forename: John C
authors.initials: JC
authors.lastname: Rogers
authors.forename: Paul
authors.initials: P
authors.lastname: Haney
authors.forename: Navid
authors.initials: N
authors.lastname: Haghdoost
mesh_terms: D000818:Animals; D002417:Cattle; D005591:Chemical Fractionation / Q000295:instrumentation*; D002853:Chromatography, Liquid; D003902:Detergents / Q000302:isolation & purification*; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D009844:Oligosaccharides / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D012117:Resins, Synthetic / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013499:Surface Properties; D014357:Trypsin / Q000737:chemistry
chemical_list: D003902:Detergents; D009844:Oligosaccharides; D010455:Peptides; D011506:Proteins; D012117:Resins, Synthetic; D012710:Serum Albumin, Bovine; D014357:Trypsin