TITLE

Optimization of capillary electrophoresis conditions for a glucagon competitive immunoassay using response surface methodology

AUTHOR(S)
Lomasney, Anna R.; Guillo, Christelle; Sidebottom, Ashley M.; Roper, Michael G.
PUB. DATE
May 2009
SOURCE
Analytical & Bioanalytical Chemistry;May2009, Vol. 394 Issue 1, p313
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
The capillary electrophoresis (CE) conditions for a competitive immunoassay of glucagon were optimized for highest sensitivity of the immunoassay and resolution of the electrophoretic peaks using a Box–Behnken design. Injection time, voltage ramp time, and separation voltage were varied between three levels and two responses, bound-to-free ( B/ F) ratio of the immunoassay peaks and resolution between the peaks, were measured. Analysis of variance was applied to fit a predictive model, and a desirability function was used to simultaneously optimize both responses. A 10-s injection, 1.6-min ramp time, and a 22-kV separation voltage were the conditions found when high B/ F was given more emphasis than high resolution. To test the model, calibration curves of a glucagon immunoassay were measured at the optimum and least optimum CE conditions. Optimal conditions increased the sensitivity of the immunoassay by 388% compared to the least optimum conditions while maintaining adequate resolution.
ACCESSION #
37371694

 

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