Picomolar analysis of proteins using electrophoretically mediated microanalysis and capillary electrophoresis with laser-induced fluorescence detection

In Ho Lee, Devanand Pinto, Edgar A. Arriaga, Zheru Zhang, Norman J. Dovichi

Research output: Contribution to journalArticlepeer-review

106 Scopus citations

Abstract

We report a method for the analysis of picomolar concentration proteins using electrophoretically mediated microanalysis (EMMA) to label proteins on-column with a fluorogenic reagent. Labeling is followed by capillary zone electrophoresis separation and postcolumn detection based on laser-induced fluorescence. The method provides a concentration detection limit (3σ) of 3 × 10-13 M for conalbumin. The method provides separation efficiency of 300 000 theoretical plates. Protein extract from a human colon adenocarcinoma cell line generated a dozen major components and many minor components in a 12-min separation; the protein extract from 2.5 cells was used for this analysis. When compared to UV absorbance detection, the EMMA method provides 7 000 000-fold improvement in detection limit.

Original languageEnglish (US)
Pages (from-to)4546-4548
Number of pages3
JournalAnalytical chemistry
Volume70
Issue number21
DOIs
StatePublished - Nov 1 1998

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