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Transient transfection and expression of eukaryotic membrane proteins in Expi293F cells and their screening on a small scale: Application for structural studies

DOI: 10.1007/978-1-0716-1406-8_5 DOI Help

Authors: Ganna O. Krasnoselska (The Wellcome Centre for Human Genetics, University of Oxford) , Maud Dumoux (The Rosalind Franklin Institute) , Nadisha Gamage (Diamond Light Source) , Harish Cheruvara (Diamond Light Source) , James Birch (Diamond Light Source) , Andrew Quigley (Diamond Light Source) , Raymond J. Owens (The Wellcome Centre for Human Genetics, University of Oxford; The Rosalind Franklin Institute)
Co-authored by industrial partner: No

Type: Book Chapter
ISBN: 978-1-0716-1406-8
Book Chapter: 5

State: Published (Approved)
Published: May 2021

Abstract: Cancers, neurodegenerative and infectious diseases remain some of the leading causes of deaths worldwide. The structure-guided drug design is essential to advance drug development for these important diseases. One of the key challenges in the structure determination workflow is the production of eukaryotic membrane proteins (drug targets) of high quality. A number of expression systems have been developed for the production of eukaryotic membrane proteins. In this chapter, an optimized detailed protocol for transient transfection and expression of eukaryotic membrane proteins in Expi293F cells is presented. Testing expression and purification on a small scale allow optimizing conditions for sample preparation for downstream structural (cryo-EM) elucidation.

Journal Keywords: Protein purification; Mammalian expression system; Expi293F cells; Transient expression; Membrane proteins; GPCR; Detergent screen; Small-scale tests; FSEC

Subject Areas: Biology and Bio-materials, Medicine, Technique Development


Technical Areas:

Added On: 26/05/2021 11:33

Discipline Tags:

Life Sciences & Biotech Health & Wellbeing Drug Discovery Structural biology Technique Development - Life Sciences & Biotech

Technical Tags: