title: On-line hollow-fiber flow field-flow fractionation-electrospray ionization/time-of-flight mass spectrometry of intact proteins.
abstract: Capabilities of mass spectrometry for the analysis of intact proteins can be increased through separation methods. Flow field-flow fractionation (FlFFF) is characterized by the particularly "soft" separation mechanism, which is ideally suited to maintain the native structure of intact proteins. This work describes the original on-line coupling between hollow-fiber FlFFF (HF FlFFF), the microcolumn variant of FlFFF, and electrospray ionization/time-of-flight mass spectrometry (ESI/TOFMS) for the analysis and characterization of intact proteins. The results show that the native (or pseudonative) structure of horse heart myoglobin and horseradish peroxidase is maintained. Sample desalting is also observed for horse heart myoglobin. Correlation between the molar mass values independently measured by HF FlFFF retention and ESI/TOFMS allows us to confirm the protein aggregation features of bovine serum albumin and to indicate possible changes in the quaternary structure of human hemoglobin.
authors.affiliation: Department of Chemistry G. Ciamician, University of Bologna, Via Selmi 2, I-40126 Bologna, Italy.
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Reschiglian
authors.forename: Andrea
authors.initials: A
authors.lastname: Zattoni
authors.forename: Barbara
authors.initials: B
authors.lastname: Roda
authors.forename: Leonardo
authors.initials: L
authors.lastname: Cinque
authors.forename: Daniela
authors.initials: D
authors.lastname: Parisi
authors.forename: Aldo
authors.initials: A
authors.lastname: Roda
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Dal Piaz
authors.forename: Myeong Hee
authors.initials: MH
authors.lastname: Moon
authors.forename: Byung Ryul
authors.initials: BR
authors.lastname: Min
mesh_terms: D002851:Chromatography, High Pressure Liquid; D031521:Fractionation, Field Flow / Q000379:methods*; D006454:Hemoglobins / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D009211:Myoglobin / Q000737:chemistry; D020836:Protein Structure, Quaternary; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D006454:Hemoglobins; D009211:Myoglobin; D011506:Proteins; D012710:Serum Albumin, Bovine; D006735:Horseradish Peroxidase