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Efficient Mammalian Cell Expression and Single-step Purification of Extracellular Glycoproteins for Crystallization

Article DOI: 10.3791/53445-v • 07:08 min • December 23rd, 2015
December 23rd, 2015 •
*1,2,3, *2,3,4,5, 2,3,5,6,7
1Molecular Microbiology and Microbial Pathogenesis Program, Washington University School of Medicine, 2Department of Internal Medicine, Washington University School of Medicine, 3Drug Discovery Program in Pulmonary and Critical Care Medicine, Washington University School of Medicine, 4Biochemistry Program, Washington University School of Medicine, 5Center for the Investigation of Membrane Excitability Diseases, Washington University School of Medicine, 6Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 7Department of Cell Biology and Physiology, Washington University School of Medicine
* These authors contributed equally

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This is a quick, cost-efficient protocol for the production of secreted, glycosylated mammalian proteins and subsequent single-step purification with sufficient yields of homogenous protein for X-ray crystallography and other biophysical studies.

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Mammalian Cell Expression Extracellular Glycoproteins Crystallization Secreted Protein Expression Purification Nickel Affinity Chromatography Transient Transfection Suspension Culture Transfection Agents Glucose Levels Native Glycans Protein Yield X-ray Crystallography Single-step Purification
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