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Picomolar assay of native proteins by capillary electrophoresis precolumn labeling, submicellar separation, and laser-induced fluorescence detection

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1021/ac9611677
AuthorSearch for: 1ORCID identifier: https://orcid.org/0000-0002-0665-7041; Search for: 1; Search for: 1; Search for: 1; Search for: 1; Search for: 1; Search for: 1; Search for: 2
Affiliation
  1. Department of Chemistry, University of Alberta
  2. Defense Research Establishment Suffield
FormatText, Article
Subjectfluorescence; labeling; monomers; peptides and proteins; sodium dodecyl sulfate
Abstract
We report a method for the assay of proteins at concentrations lower than 10⁻¹⁰ M with as little as 200 amol of protein. High sensitivity is accomplished by derivatizing the ε-amino group of the protein's lysine residues with the fluorogenic dye 5-furoylquinoline-3-carboxaldehyde and use of a sheath flow cuvette fluorescence detector. Most proteins have a large number of lysine residues; therefore, a large number of fluorescent molecules can be attached to each protein molecule. In general, precolumn labeling improves sensitivity but degrades resolution due to the inhomogeneity of the reaction products from multiple labeling. However, we demonstrate that, through careful manipulation of the separation and reaction conditions, high sensitivity can be obtained without excessive loss in separation efficiency. Over 190 000 theoretical plates are obtained for fluorescently labeled ovalbumin.
Publication date
PublisherAmerican Chemical Society
In
  • Analytical Chemistry 69, no. 15 (1 August 1997): 3015–3021.
LanguageEnglish
Peer reviewedYes
NRC publication
This is a non-NRC publication

"Non-NRC publications" are publications authored by NRC employees prior to their employment by NRC.

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Record created2024-07-16
Record modified2024-07-16
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