Protocol

Monoclonal Antibodies

Volume 378 of the series Methods in Molecular Biology pp 33-52

Soluble Fc Fusion Proteins for Biomedical Research

  • Meg L. FlanaganAffiliated withDepartment of Pathology, Keck School of Medicine, University of Southern California
  • , Robyn S. AriasAffiliated withDepartment of Pathology, Keck School of Medicine, University of Southern California
  • , Peisheng HuAffiliated withDepartment of Pathology, Keck School of Medicine, University of Southern California
  • , Leslie A. KhawliAffiliated withDepartment of Pathology, Keck School of Medicine, University of Southern California
  • , Alan L. EpsteinAffiliated withDepartment of Pathology, Keck School of Medicine, University of Southern California

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Abstract

As a source of recombinant antigen, soluble constant fragment (Fc) fusion proteins have become valuable reagents for immunotherapy and laboratory investigations. Additional applications for these reagents include flow cytometry, immunohistochemistry, and in vitro activity assays. To aid investigators in the generation of these reagents, the materials and methods required for producing Fc fusion proteins are described. The investigator’s protein moiety of interest is genetically linked to the N-terminus of murine Fc and subsequently expressed in large quantity using a mammalian cell expression system. The resulting Fc fusion proteins are purified on a protein A column and may be stored for at least one year at −20°C. The availability of easily purified, soluble Fc fusion proteins in such quantity can facilitate research in multiple fields of medicine and biotechnology.

Key Words

Antibody engineering antibody production soluble Fc fusion protein recombinant antigen genetic engineering protein expression protein moiety mammalian expression system