title: HPLC enrichment/isolation of proteins for post-translational modification studies from complex mixtures.
abstract: The paper describes a macroporous RP-HPLC method for separation and isolation/enrichment of proteins from complex mixtures. The method is robust and efficient; using 2.1 or 4.6mm diameter columns provides sufficient material for subsequent proteomic analysis. The main advantage of the method is that most protein variants are isolated in the same fraction, as separation is not based on differences in isoelectric point. This is highly advantageous for studying complex mixtures and post-translational modifications. Examples related to glycosylation analysis are discussed in detail.
authors.affiliation: Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar tudósok körútja 2., Budapest 1117, Hungary.
authors.forename: Eszter
authors.initials: E
authors.lastname: Tóth
authors.affiliation: Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar tudósok körútja 2., Budapest 1117, Hungary.
authors.forename: Olivér
authors.initials: O
authors.lastname: Ozohanics
authors.affiliation: Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar tudósok körútja 2., Budapest 1117, Hungary; Budapest University of Technology and Economics, Department of Inorganic and Analytical Chemistry, Szt. Gellért tér 4., Budapest 1111, Hungary.
authors.forename: Balázs
authors.initials: B
authors.lastname: Bobály
authors.affiliation: Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar tudósok körútja 2., Budapest 1117, Hungary.
authors.forename: Ágnes
authors.initials: Á
authors.lastname: Gömöry
authors.affiliation: Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar tudósok körútja 2., Budapest 1117, Hungary.
authors.forename: Anita
authors.initials: A
authors.lastname: Jekő
authors.affiliation: Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar tudósok körútja 2., Budapest 1117, Hungary.
authors.forename: László
authors.initials: L
authors.lastname: Drahos
authors.affiliation: Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar tudósok körútja 2., Budapest 1117, Hungary. Electronic address: vekey.karoly@ttk.mta.hu.
authors.forename: Károly
authors.initials: K
authors.lastname: Vékey
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D045424:Complex Mixtures / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D010949:Plasma / Q000737:chemistry; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics / Q000379:methods
chemical_list: D045424:Complex Mixtures; D011506:Proteins
keywords: Glycosylation; HPLC; Mass spectrometry; Proteins; Sample preparation