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Title

Multiple peaks in HPLC of proteins : Bovine serum albumin eluted in a reversed-phase system

AuthorsFrutos, Mercedes de; Cifuentes, Alejandro ; Diez-Masa, Jose Carlos; Camafeita, Emilio; Méndez Cormán, Enrique
KeywordsReversed-phase HPLC
capillary electrophoresis
MALDI-MS
Whey Proteins
Multiple Peaks
Bovine Serum Albumin
Issue Date1998
PublisherJohn Wiley & Sons
CitationJournal of High Resolution Chromatography 21 (1) : 18–24 (1998)
AbstractElution of a commercial sample of bovine serum albumin (BSA) in a reversed-phase (RP) HPLC system at room temperature gives a distorted peak. If a shallow gradient is used during elution a split peak is observed. The nature of the several parts of this multiple peak is studied using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), size-exclusion chromatography (SEC), amino acid analysis, re-elution in RP-HPLC of collected fractions, capillary electrophoresis (CE), and matrix assisted laser desorption ionization-mass spectrometry (MALDI-MS). This study demonstrates that the split peak of BSA observed in these chromatographic conditions is due to the monomer, dimer and other aggregates existing in the commercial sample of the BSA used. Moreover, it is proved that typical RP-chromatographic conditions do not cause aggregation of BSA.
Publisher version (URL)http://dx.doi.org/10.1002/(SICI)1521-4168(19980101)21:1<18::AID-JHRC18>3.0.CO;2-5
URIhttp://hdl.handle.net/10261/43146
DOI10.1002/(SICI)1521-4168(19980101)21:1<18::AID-JHRC18>3.0.CO;2-5
ISSN0935-6304
Appears in Collections:(IQOG) Artículos
(CNB) Artículos
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