Tools for Coproducing Multiple Proteins in Mammalian Cells

  • Zahra Assur
  • Wayne A. Hendrickson
  • Filippo ManciaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 801)


Structural and functional studies of many mammalian systems are critically dependent on abundant supplies of recombinant multiprotein complexes. Mammalian cells are often the most ideal, if not the only suitable host for such experiments. This is due to their intrinsic capability to generate functional mammalian proteins. This advantage is frequently countered by problems with yields in expression, time required to generate overexpressing lines, and elevated costs. Coexpression of multiple proteins adds another level of complexity to these experiments, as cells need to be screened and selected for expression of suitable levels of each component. Here, we present an efficient fluorescence marking procedure for establishing stable cell lines that overexpress two proteins in coordination, and we validate the method in the production of recombinant monoclonal antibody Fab fragments. This procedure may readily be expanded to systems of greater complexity, comprising more than two components.

Key words

Coexpression Monoclonal antibody Fab fragments Protein complexes Protein–protein interactions Mammalian cells 



We are grateful to Richard Axel for discussions and to Ira Schieren for sharing with us his invaluable expertise on cell sorting. This work was supported in part by NIH grants GM68671 and GM75026.


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Zahra Assur
    • 1
  • Wayne A. Hendrickson
    • 1
  • Filippo Mancia
    • 2
    Email author
  1. 1.Department of Biochemistry and Molecular BiophysicsColumbia UniversityNew YorkUSA
  2. 2.Department of Physiology and Cellular BiophysicsColumbia UniversityNew YorkUSA

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