Impurity profiling quality control testing of synthetic peptides using liquid chromatography-photodiode array-fluorescence and liquid chromatography-electrospray ionization-mass spectrometry: The obestatin case

被引:55
作者
De Spiegeleer, Bart [1 ]
Vergote, Valentijn [1 ,2 ]
Pezeshki, Adel [1 ,3 ]
Peremans, Kathelijne [2 ]
Burvenich, Christian [2 ,3 ]
机构
[1] Univ Ghent, Fac Pharmaceut Sci, Drug Qual & Registrat Grp, B-9000 Ghent, Belgium
[2] Univ Ghent, Fac Vet Med, Dept Med Imaging, B-9820 Merelbeke, Belgium
[3] Univ Ghent, Fac Vet Med, Dept Physiol & Biometr, B-9820 Merelbeke, Belgium
关键词
obestatin GPR39 binding; peptide quality; impurity profile; synthesis impurities; HPLC-PDA; fluorescence; ESI-ion trap MS/MS;
D O I
10.1016/j.ab.2008.02.014
中图分类号
Q5 [生物化学];
学科分类号
071010 [生物化学与分子生物学]; 081704 [应用化学];
摘要
Following several conflicting publications, the inability to reproduce the original findings on in vitro obestatin binding and activation of GPR39 receptors was recently reported by its discoverers, and several hypotheses to rationalize these findings were proposed. Based on one of these postulations (i.e., presence of impurities), peptide identity and impurity profiles were thoroughly evaluated on obestatin peptides obtained from five different manufacturers, as used by the different research groups. We found that one of the products examined was in reality a totally different peptide and that the quality of two-thirds of the other peptides was insufficient for in vitro and in vivo experiments (i.e., peptide purity less than 95% and/or individual impurities exceeding 1%). These observations question the divergent conclusions reported in the literature about the activity of obestatin. Therefore, we strongly recommend appropriate quality control testing before using any peptides for biomedical research purposes. (C) 2008 Elsevier Inc. All rights reserved.
引用
收藏
页码:229 / 234
页数:6
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