Evaluation of three expanded bed adsorption anion exchange matrices with the aid of recombinant enhanced green fluorescent protein overexpressed in Escherichia coli

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
C CabanneX Santarelli

Abstract

Three anion exchanger expanded bed adsorption (EBA) matrices: Streamline DEAE, Streamline Q XL and Q Hyper Z were evaluated with the aid of EFGP from an ultrasonic homogenate of Escherichia coli. Two pH of buffer were tested. Capture was done in an expanded mode whereas elution was done in a packed mode. The same conditions were chosen for evaluation of the three matrices. We observed a loss of EGFP (8-15%) in the through flow fraction especially with the Streamline Q XL matrix, probably due to an aggregation of beads during sample application. The beads of this matrix possess tentacles which probably retain a lot of cellular and molecular debris. The two other matrices gave a good purification of the EGFP (7-15-fold) but the Q Hyper Z matrix appeared to give the best results. It is composed of little size and density beads which lead to a higher exchange surface and then a better mass transfer.

References

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Oct 23, 1997·Nature Biotechnology·T Misteli, D L Spector
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Feb 1, 2002·Journal of Chromatography. a·Xiao-Dong Tong, Yan Sun
Mar 26, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·W DieryckX Santarelli

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Citations

Aug 8, 2008·Journal of Chromatography. a·A ArpanaeiT J Hobley
Feb 22, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·C CabanneX Santarelli
May 10, 2006·Protein Expression and Purification·Gabriel D PeckhamWei Wen Su
Jun 27, 2013·Molecular Membrane Biology·Suzan Ruehrer, Hartmut Michel

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