Quantitation of CRM197 using imaged capillary isoelectric focusing with fluorescence detection and capillary Western.

John W Loughney, Sha Ha, Richard R Rustandi

Analytical biochemistry 2017 Oct 01

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Maurice is a new instrument that can perform imaged capillary isoelectric focusing (icIEF). The standard detection for icIEF is UV absorbance at 280 nm, which limits its application to high protein concentration samples and non-complex samples. Here we describe an icIEF instrument with fluorescence detection. We demonstrate the advantage of using either icIEF with fluorescence detection or quantitative Western Blot to measure diphtheria toxin mutant CRM197 protein titer in crude cell lysates and purified samples. These two techniques have great potentials to become standard methods to analyze protein titers in crude cell lysate or other complex samples types. Copyright © 2017 Elsevier Inc. All rights reserved.

Citation

John W Loughney, Sha Ha, Richard R Rustandi. Quantitation of CRM197 using imaged capillary isoelectric focusing with fluorescence detection and capillary Western. Analytical biochemistry. 2017 Oct 01;534:19-23