Application of electrospray mass spectrometry to the characterization of recombinant proteins up to 44 kDa.
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.
Van Dorsselaer
A
A
Laboratoire de Chimie Organique des Substances Naturelles, URA31 CNRS, 67084 Strasbourg, France.
Bitsch
F
F
Green
B
B
Jarvis
S
S
Lepage
P
P
Bischoff
R
R
Kolbe
V J
VJ
Roitsch
C
C
D000595:Amino Acid Sequence; D006678:HIV / Q000032:analysis; D006629:Hirudins / Q000032:analysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet; D014764:Viral Proteins / Q000032:analysis; D000515:alpha 1-Antitrypsin / Q000032:analysis
D006629:Hirudins; D011994:Recombinant Proteins; D014764:Viral Proteins; D000515:alpha 1-Antitrypsin