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

被引：20

作者：

Loughney, John W. ^{[1
]}

Ha, Sha ^{[1
]}

Rustandi, Richard R. ^{[1
]}

机构：

[1] Merck & Co Inc, Biol & Vaccines, Merck Res Labs, Kenilworth, NJ USA

来源：

ANALYTICAL BIOCHEMISTRY | 2017年 / 534卷

关键词：

Maurice; Imaged capillary IEF; Fluorescent detection; CRM197; Wes; Western blot; BLOT; PROTEINS; VACCINE; SYSTEM; CE;

D O I：

10.1016/j.ab.2017.06.013

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (C) 2017 Elsevier Inc. All rights reserved.

引用

页码：19 / 23

页数：5

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