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A rapid method for removal of detergents from proteins and protein digests in a spin column format
Author(s) -
Antharavally Babu S,
Salunkhe Ashok M,
Mallia Krishna A,
Rosenblatt Michael M,
Rogers John C,
Haghdoost Navid R,
Haney Paul J,
Benton Betsy,
Bell Peter A
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.519.1
Subject(s) - chromatography , chemistry , trypsin , chaps , octyl glucoside , biochemistry , enzyme
We have developed a high‐performance resin that offers exceptional detergent removal for proteins and peptides. The easy‐to‐use spin format dramatically improves results over standard drip column or batch methodologies with >95% removal from 1–5% solutions of detergents such as 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. Post‐column LC‐MS/MS analysis of BSA trypsin digests followed by analysis of the data using MASCOT revealed excellent sequence coverage, and the high MASCOT scores obtained indicate successful removal of detergent from the digested peptides. MALDI‐MS analysis of unprocessed and processed samples further confirmed efficient removal of detergents. The advantages of this method include speed (less than 15 minutes), efficient detergent removal, and high recovery of proteins/peptides.