[LC/MS research on the isomers of special melanoma antigen-encoding gene-2 epitope peptide induced by solvent].
To study the influence of different solvent systems on the isomers of melanoma antigen-encoding gene-2(MAGE-2) epitope peptide synthesized by Marrifield's solid synthesis method, MAGE-2(171-179) epitope peptides were pre-treated using ethanol and methanol systems respectively, and then analysed by RP-HPLC/MS, with dimethyl-sulphoxide (DMSO) as control solvent. Results demonstrated that 100% ethanol and 100% methanol solvents could induce the production of isomers of MAGE-2, while 100% DMSO, 50% ethanol and 50% methanol could not. These isomeric peptides could be reversed greatly if treated with trifluoroacetic acid (TFA). The results also showed that the analysis would be distorted and the yield of peptides would decrease dramatically if the peptides were pre-treated with high concentrations of ethanol and methanol before RP-HPLC analysis and preparation. Thus it is suggested that it might be better to decrease the concentration of organic phase in the solvent system for the resolvable hydrophobic peptides, acidified ethanol and methanol might also be better for the highly hydrophobic peptides, and DMSO would be a good solvent for hydrophobic peptides analysis provided that it would not influence the resolving power of chromatography.
Zhou
W
W
Institute of Immunology PLA, Third Military Medical University, Chongqing 400038, China. weizhou@mail.tmmu.com.cn
Wu
Y Z
YZ
Bian
J
J
Jia
Z C
ZC
Tang
Y
Y
Zou
L Y
LY
D000951:Antigens, Neoplasm / Q000235:genetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004121:Dimethyl Sulfoxide; D000939:Epitopes; D008401:Gas Chromatography-Mass Spectrometry; D009363:Neoplasm Proteins / Q000032:analysis / Q000235:genetics*; D010455:Peptides; D012997:Solvents; D013237:Stereoisomerism; D014269:Trifluoroacetic Acid
D000951:Antigens, Neoplasm; D000939:Epitopes; D009363:Neoplasm Proteins; D010455:Peptides; D012997:Solvents; D014269:Trifluoroacetic Acid; D004121:Dimethyl Sulfoxide