Literature DB >> 28666777

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

John W Loughney1, Sha Ha1, Richard R Rustandi2.   

Abstract

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.

Keywords:  CRM197; Fluorescent detection; Imaged capillary IEF; Maurice; Wes; Western blot

Mesh:

Substances:

Year:  2017        PMID: 28666777     DOI: 10.1016/j.ab.2017.06.013

Source DB:  PubMed          Journal:  Anal Biochem        ISSN: 0003-2697            Impact factor:   3.365


  4 in total

1.  Platform Methods to Characterize the Charge Heterogeneity of Three Common Protein Therapeutics by Imaged Capillary Isoelectric Focusing.

Authors:  Jiaqi Wu; Will McElroy; Charles Haitjema; Carsten Lück; Christopher Heger
Journal:  Methods Mol Biol       Date:  2021

2.  Fast Immunoassay for Microfluidic Western Blotting by Direct Deposition of Reagents onto Capture Membrane.

Authors:  Natalie E Arvin; Mohamed Dawod; Don T Lamb; Jon P Anderson; Michael D Furtaw; Robert T Kennedy
Journal:  Anal Methods       Date:  2020-03-04       Impact factor: 2.896

Review 3.  Protein purification and analysis: next generation Western blotting techniques.

Authors:  Manish Mishra; Shuchita Tiwari; Aldrin V Gomes
Journal:  Expert Rev Proteomics       Date:  2017-10-13       Impact factor: 3.940

4.  Development of an imaged capillary isoelectric focusing method for characterizing the surface charge of mRNA lipid nanoparticle vaccines.

Authors:  John W Loughney; Kevin Minsker; Sha Ha; Richard R Rustandi
Journal:  Electrophoresis       Date:  2019-07-24       Impact factor: 3.535

  4 in total

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