title: Reversed phase liquid chromatography hyphenated to continuous flow-extractive desorption electrospray ionization-mass spectrometry for analysis and charge state manipulation of undigested proteins.
abstract: The application of continuous flow-extractive desorption electrospray ionization (CF-EDESI), an ambient ionization source demonstrated previously for use with intact protein analysis, is expanded here for the coupling of reversed phase protein separations to mass spectrometry. This configuration allows the introduction of charging additives to enhance detection without affecting the chromatographic separation mechanism. Two demonstrations of the advantages of CF-EDESI are presented in this work. First, a proof-of- principle is presented to demonstrate the applicability of hyphenation of liquid chromatography (LC) to CF- EDESI. LC-CF-EDESI-MS has good sensitivity compared to LC-electrospray ionization (ESI)-mass spectrometry. Second, the supercharging mechanism investigated in CF-EDESI provides an insight into a highly debated supercharging process in ESI. The results indicate that the mechanism of protein charging seen in HPLC-CF-EDESI is different from supercharging phenomena in conventional ESI. The surface tension mechanism and binding mechanism may both contribute to protein supercharging in ESI.
authors: Department of Chemistry & Biochemistry, the University of Texas at Arlington, Arlington, Texas, USA. lili@uta.edu.
authors: Li
authors: L
authors: Li
authors: Department of Chemistry & Biochemistry, the University of Texas at Arlington, Arlington, Texas, USA.. yang.samuel@gene.com.
authors: Samuel H
authors: SH
authors: Yang
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220, Prague, Czech Republic.. veronika.vidova@gmail.com.
authors: Veronika
authors: V
authors: Vidova
authors: Department of Chemistry & Biochemistry, the University of Texas at Arlington, Arlington, Texas, USA.. elisa.rice@mavs.uta.edu.
authors: Elisa M
authors: EM
authors: Rice
authors: Department of Chemistry & Biochemistry, the University of Texas at Arlington, Arlington, Texas, USA.. roger.giles@srlab.co.uk.
authors: Aruna B
authors: AB
authors: Wijeratne
authors: Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220, Prague, Czech Republic. Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacky University, 17.listopadu 12, 771 46 Olomouc, Czech Republic.. vlhavlic@biomed.cas.cz.
authors: Vladimír
authors: V
authors: Havlíček
authors: Department of Chemistry & Biochemistry, the University of Texas at Arlington, Arlington, Texas, USA.. kschug@uta.edu.
authors: Kevin A
authors: KA
authors: Schug
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D056148:Chromatography, Reverse-Phase / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D010449:Peptide Mapping / Q000295:instrumentation*; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D013048:Specimen Handling / Q000295:instrumentation*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation*; D055672:Static Electricity
chemical_list: D011506:Proteins