Protein analysis using capillary electrophoresis coupled to mass spectrometry through vibrating sharp‐edge spray ionization

Author:

Witzel Makenzie T1ORCID,Veltri Lindsay M.1,Kostelic Marius2,Elshamy Yousef S.1,Lucas John A.1,Lai Stella2,Du Chen2,Wysocki Vicki H.2,Holland Lisa A.1ORCID

Affiliation:

1. C. Eugene Bennett Department of Chemistry West Virginia University Morgantown West Virginia USA

2. Resource for Native Mass Spectrometry Guided Structural Biology The Ohio State University Columbus Ohio USA

Abstract

AbstractCapillary electrophoresis (CE) interfaced to mass spectrometry (MS) with electrospray ionization typically incorporates acidic additives or organic solvents to assist in ionization. Vibrating sharp‐edge spray ionization (VSSI) is a voltage‐free method to interface CE and MS that does not require these additives, making it appealing for protein analyses. CE–VSSI nanoflow sheath separations are performed with low ionic strength aqueous solutions in the sheath to reduce suppression. Serine is also included in the sheath to reduce analyte adduction. Proteins are detected in the 2.5–10 µM range, corresponding to an injected mass range of 0.1–1.2 ng. The anionic proteins β‐lactoglobulin and transferrin are resolved using an unmodified fused silica capillary because they do not exhibit nonspecific surface adsorption. Conversely, separations of cationic proteins cytochrome c, ribonuclease A, and α‐chymotrypsinogen A in an unmodified capillary require acidic background electrolytes to overcome adsorption. Alternatively, a semipermanent coating comprised self‐assembled lipids overcomes surface adsorption at a neutral pH. Separations with zwitterionic and hybrid cationic coatings are complete within 15 or 6 min, respectively. The dimeric form of triosephosphate isomerase was observed at a 60 µM, corresponding to a mass of 19 ng, by dropping the temperature of the MS inlet.

Funder

National Institutes of Health

Publisher

Wiley

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