Novel chip-based isoelectric focusing device for fractionation of bacteria prior to their mass spectrometry identification

0551185 - ÚIACH 2023 RIV NL eng J - Journal Article
Duša, Filip - Šalplachta, Jiří - Horká, Marie - Lunerová, K. - Rosenbergová, K. - Kubíček, O.
Novel chip-based isoelectric focusing device for fractionation of bacteria prior to their mass spectrometry identification.
Analytica Chimica Acta. Roč. 1192, FEB (2022), č. článku 339333. ISSN 0003-2670. E-ISSN 1873-4324
R&D Projects: GA ČR(CZ) GA19-00742S; GA MV VI20172020069
Institutional support: RVO:68081715
Keywords : isoelectric focusing * chip * mass spectrometry
OECD category: Analytical chemistry
Impact factor: 6.2, year: 2022
Method of publishing: Limited access

We have developed a planar chip utilizing divergent geometry of separation channel capable of vertical free-flow electrophoresis of particles at flows of lower hundreds of microliters per minute. The divergent flow isoelectric focusing (DF-IEF) chip consists of two sheets of clear polystyrene glass which serve as a base with working channels and a top cover sealing the separation channel. Optimization showed that the chip is capable to form pH gradient within 1 h and separation is completed in 5 or more minutes depending on the sample volume. The vertical position of the chip enabled analysis of sedimenting particles including microorganisms. Four different common bacteria species inactivated with H2O2 vapors were analyzed in a series of experiments. Isoelectric points were determined with capillary isoelectric focusing with following fractionation using DF-IEF with intact cell matrix-assisted laser desorption/ionization mass spectrometry detection. The DF-IEF chip fractionation proved promising for bacterial sample preparation from complex matrices for subsequent identification of whole cells by mass spectrometry.
Permanent Link: http://hdl.handle.net/11104/0326625