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Isolation and Characterization Of Chimeric Human Fc-expressing Proteins Using Protein A Membrane Adsorbers And A Streamlined Workflow
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Construction, expression, and purification of chimeric protein reagents based on immunoglobulin fc regions

John P Cannon1, Marci O'Driscoll, Gary W Litman

  • 1Department of Pediatrics, University of South Florida College of Medicine, University of South Florida and All Children's Hospital Children's Research Institute, St. Petersburg, FL, USA. jcannon@health.usf.edu

Methods in Molecular Biology (Clifton, N.J.)
|June 25, 2011
PubMed

Recombinant fusion proteins incorporating experimental protein domains fused to immunoglobulin Fc regions have become widely utilized in studies of protein-ligand interactions. The advantages of these systems include an inherent increase in avidity provided by the multimerization of Fc regions, combined with robust detection methods based on numerous commercially available secondary reagents directed against the Fc tag. We describe a set of methods for subcloning, expression, and purification of chimeric protein reagents containing a protein domain (or domains) of interest fused to a C-terminal moiety derived from the Fc region of either IgG or IgM.

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