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

doi:10.1021/ac980360t

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

Chemistry (Twin Cities)

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110 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 language	English (US)
Pages (from-to)	4546-4548
Number of pages	3
Journal	Analytical chemistry
Volume	70
Issue number	21
DOIs	https://doi.org/10.1021/ac980360t
State	Published - Nov 1 1998

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

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.",

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T1 - Picomolar analysis of proteins using electrophoretically mediated microanalysis and capillary electrophoresis with laser-induced fluorescence detection

AU - Lee, In Ho

AU - Pinto, Devanand

AU - Arriaga, Edgar A.

AU - Zhang, Zheru

AU - Dovichi, Norman J.

PY - 1998/11/1

Y1 - 1998/11/1

N2 - 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.

AB - 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.

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

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