Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10560147 | Talanta | 2011 | 5 Pages |
Abstract
A new method based on solid-support reaction is described to realize fluorescent derivatization of proteins at concentrations as low as 10â8Â M. A simple, low-cost homemade capillary C18 cartridge was fabricated as the solid-support reactor. Using bovine serum albumin (BSA) as a test protein, we demonstrated that the protein can be captured by this reactor and then labeled by fluorescein isothiocyanate (FITC, isomer I) on solid-support. Unwanted fluorescent intruder (excrescent FITC and products of secondary reactions) were removed from target easily. The analysis by nano-HPLC with laser-induced fluorescence (LIF) detection was described. The effect of reaction conditions on the derivatization has been evaluated and discussed. The use of the solid-support reactor allows easy handling of as little as 8.5Â pmol of BSA. A fraction from weak anion-exchange chromatography (WAX) of human liver extract was used as an illustrative example of application to real samples.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Qian Tao, Ming-Xia Gao, Guang-Feng Hong, Qi Chen, Xiang-Min Zhang,