Protein tryptic digests analyzed by carrier ampholyte-based capillary electrophoresis coupled to ESI-MS.
In this study, narrow pH cuts of carrier ampholytes have been used as buffers in CE for the analysis of protein tryptic digests. Their low conductivity allows very efficient separations under high electric field strength without inducing any significant Joule heating. In this study, the capabilities of narrow pH cuts of carrier ampholytes for the separation of protein tryptic digests have been assessed. Three proteins of different molecular masses have been studied: cytochrome C (horse heart), beta-lactoglobulin B (bovine) and human transferrin. Efficient, rapid and repeatable separations of the peptides resulting from the tryptic digestion have been achieved in this buffer. Moreover, the feasibility of the coupling of carrier ampholyte-based capillary electrophoresis with ESI-MS has been demonstrated through the study of the cytochrome C tryptic digest.
Busnel
Jean-Marc
JM
Laboratory of Environment and Analytical Chemistry, UMR CNRS 7121, ESPCI, Paris, France. jean-marc.busnel@espci.fr
Descroix
Stéphanie
S
Le Saux
Thomas
T
Terabe
Shigeru
S
Hennion
Marie-Claire
MC
Peltre
Gabriel
G
D000665:Ampholyte Mixtures / Q000737:chemistry*; D000818:Animals; D045304:Cytochromes c / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000379:methods*; D006736:Horses; D010446:Peptide Fragments / Q000302:isolation & purification*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
D000665:Ampholyte Mixtures; D010446:Peptide Fragments; D045304:Cytochromes c; D014357:Trypsin