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Issue 19, 2017
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Quantification of intractable membrane proteins in genetically engineered crops by liquid chromatography coupled with tandem mass spectrometry

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Abstract

Liquid chromatography with tandem mass spectrometry (LC-MS/MS) methods to quantify a membrane protein in genetically engineered maize leaves and another in soybean seeds were developed and validated. An extraction buffer containing 100 mM ammonium bicarbonate, 0.5% CHAPS (w/v) and 0.1% Triton X-100 (v/v) was determined to be suitable. High recovery (>88%) and good reproducibility (coefficient of variance ≤ 15%) were demonstrated for both membrane proteins. Recombinant proteins were used as standards to prepare calibration curves with good linearity (correlation coefficient > 0.997) and accuracy (deviation from nominal concentration <15%). Our study demonstrates the suitability of LC-MS/MS for quantification of intractable membrane proteins in genetically engineered crops.

Graphical abstract: Quantification of intractable membrane proteins in genetically engineered crops by liquid chromatography coupled with tandem mass spectrometry

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Publication details

The article was received on 17 Jan 2017, accepted on 23 Apr 2017 and first published on 24 Apr 2017


Article type: Paper
DOI: 10.1039/C7AY00161D
Anal. Methods, 2017,9, 2821-2829

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    Quantification of intractable membrane proteins in genetically engineered crops by liquid chromatography coupled with tandem mass spectrometry

    L. J. Schacherer, M. A. Owens and T. X. Hu, Anal. Methods, 2017, 9, 2821
    DOI: 10.1039/C7AY00161D

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