Screening carbohydrate libraries for protein interactions using the direct ESI-MS assay: Applications to libraries of unknown concentration

Simple item page
dc.contributor.authorEl-Hawiet, Amr
dc.contributor.authorKlassen, John S.
dc.contributor.authorKitova, Elena N.
dc.date.accessioned2025-05-01T12:11:08Z
dc.date.available2025-05-01T12:11:08Z
dc.date.issued2014
dc.descriptionA semiquantitative electrospray ionization mass spectrometry (ESI-MS) binding assay suitable for analyzing mixtures of oligosaccharides, at unknown concentrations, for interactions with target proteins is described. The assay relies on the differences in the ratio of the relative abundances of the ligand-bound and free protein ions measured by ESI-MS at two or more initial protein concentrations to distinguish low affinity (≤103 M–1) ligands from moderate and high affinity (>105 M–1) ligands present in the library and to rank their affinities. Control experiments were performed on solutions of a single chain antibody and a mixture of synthetic oligosaccharides, with known affinities, in the absence and presence of a 40-component carbohydrate library to demonstrate the implementation and reliability of the assay. The application of the assay for screening natural libraries of carbohydrates against proteins is also demonstrated using mixtures of human milk oligosaccharides, isolated from breast milk, and fragments of a bacterial toxin and human galectin 3.
dc.identifier.doihttps://doi.org/10.7939/R3697099V
dc.language.isoen
dc.relationhttp://doi.org/10.1007/s13361-014-0964-2
dc.relation.isversionofKitova, E. N., El-Hawiet, A., & Klassen, J. S. (2014). Screening carbohydrate libraries for protein interactions using the direct ESI-MS assay: Applications to libraries of unknown concentration. Journal of the American Society for Mass Spectrometry, 25(11), 1908-1916. http://doi.org/10.1007/s13361-014-0964-2
dc.rights© 2014 Kitova, E. N., El-Hawiet, A., & Klassen, J. S. This version of this article is open access and can be downloaded and shared. The original author(s) and source must be cited.
dc.subjectCarbohydrates
dc.subjectProtein interactions
dc.subjectLibrary screening
dc.titleScreening carbohydrate libraries for protein interactions using the direct ESI-MS assay: Applications to libraries of unknown concentration
dc.typehttp://purl.org/coar/resource_type/c_6501 http://purl.org/coar/version/c_970fb48d4fbd8a85
ual.jupiterAccesshttp://terms.library.ualberta.ca/public

Files

Original bundle

Now showing 1 - 1 of 1
Thumbnail Image
Name:
JASMS_25_11_1908_8ec67ecf-a9e5-4955-a0b1-5996bbc2881d.pdf
Size:
1.12 MB
Format:
Adobe Portable Document Format
Download

Collections

Journal Articles (Chemistry)