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Solid‐phase fluorescent labeling reaction of picomole amounts of insulin in very dilute solutions and their analysis by capillary electrophoresis

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1002/elps.1150160188
AuthorSearch for: ORCID identifier: https://orcid.org/0000-0002-0665-7041; Search for: ; Search for: ; Search for: ; Search for:
FormatText, Article
Subjectfluorogenic reagents; peptide; fluorescent labeling; solid-phase
Abstract
The fluorescent labeling of peptides at concentrations as low as 10⁻⁸ M can be achieved by using a solid-phase reactor. Using oxidized insulin chain B as a test peptide, we demonstrate the use of an Immobilon CD membrane to capture and preconcentrate peptides. Insulin chain B can then be labeled with a fluorogenic reagent, 3-(2-furoyl)quinoline-2-carboxaldehyde, while it is still attached to the membrane. Unwanted fluorescent products (attributed to secondary reactions) can be washed away with methanol without significant removal of the labeled insulin chain B, which then can be extracted with a low pH buffer. The analysis by micellar electrokinetic capillary chromatography with post-column laser-induced fluorescence detection (mass limit of detection of 2.4 × 10⁻²¹ moles insulin chain B) results in electropherograms that show great improvement in terms of unwanted peaks and high number of theoretical plates (up to 20 million). The use of the solid-phase reactor allows easy handling of as little as 5 picomoles of insulin chain B.
Publication date
PublisherWiley
In
  • Electrophoresis 16, no. 1 (January 1995): 534–540.
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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