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Application of electrospray mass spectrometry to the characterization of recombinant proteins up to 44 kDa
Author(s) -
Van Dorsselaer Alain,
Bitsch Francis,
Green Brian,
Jarvis Stuart,
Lepage Pierre,
Bischoff Rainer,
Kolbe Hanno V. J.,
Roitsch Carolyn
Publication year - 1990
Publication title -
biomedical and environmental mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 0887-6134
DOI - 10.1002/bms.1200191108
Subject(s) - recombinant dna , mass spectrometry , chemistry , cysteine , dimer , molecular mass , electrospray ionization , chromatography , amino acid , electrospray , covalent bond , characterization (materials science) , biochemistry , nanotechnology , enzyme , materials science , organic chemistry , gene
Mass measurement by electrospray mass spectrometry (ESMS) is used as a rapid preliminary verification of the identity of various recombinant proteins ranging from 7 to 44 kDa with an accuracy of 0.01–0.03%. ESMS not only improves the speed but also the reliability of the protein structure determination when used in conjunction with other methods of protein analysis. Modifications of these large molecules, for example the loss of C‐terminal amino acids, N‐terminal acetylation, 2‐mercaptoethanol addition to a cysteine, and trace formation of a covalent dimer (3%), are easily detected individually or in mixtures by mass measurement using ESMS; feats which would be very difficult to achieve using classical biochemical methods. As little as 1% of several structurally related protein contaminants have been identified in a 15 kDa recombinant protein preparation.