COMPARATIVE PEPTIDE-MAPPING OF A HEPATITIS-C VIRAL RECOMBINANT PROTEIN BY CAPILLARY ELECTROPHORESIS AND MATRIX-ASSISTED LASER-DESORPTION TIME-OF-FLIGHT MASS-SPECTROMETRY
Ma. Winkler et al., COMPARATIVE PEPTIDE-MAPPING OF A HEPATITIS-C VIRAL RECOMBINANT PROTEIN BY CAPILLARY ELECTROPHORESIS AND MATRIX-ASSISTED LASER-DESORPTION TIME-OF-FLIGHT MASS-SPECTROMETRY, Journal of chromatography, 744(1-2), 1996, pp. 177-185
Citations number
10
Categorie Soggetti
Chemistry Analytical","Biochemical Research Methods
Capillary electrophoresis (CE) and matrix-assisted laser desorption ti
me-of-flight mass spectrometry (MALDI-TOF-MS) were investigated as alt
ernatives to sodium dodecyl sulfate (SDS)-polyacrylamide gel electroph
oresis for peptide mapping with Staphylococcus aureus protease (V8) of
a hydrophobic recombinant hepatitis C virus antigen, HC-31, which req
uired 0.1% SDS for solubility. Controls (V8 only) or HC-31 digests wer
e extracted with chloroform-methanol-water (1:4:3) to remove SDS, whic
h interferes with MALDI-TOF, and high salt content, which affects CE.
In two different runs by CE, the elution times of each of 11 peptide p
eaks were very reproducible (R.S.D.<0.016). 25 fragments were resolved
by MALDI-TOF-MS, including six smaller peptides (M(r)<13 000) resulti
ng from V8 autodigestion. MALDI-TOF-MS indicated that partial cleavage
s occurred, primarily at sites where there are paired glutamic and/or
aspartic acid residues.