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Mammalian Cell Expression of Single–Chain Fv (sFv) Antibody Proteins and Their C–terminal Fusions with Interleukin–2 and Other Effector Domains

Bio/Technology volume 12, pages 890–897 (1994)Cite this article

Abstract

The production of several single–chain Fv (sFv) antibody proteins was examined by three modes of mammalian cell expression. Our primary model was the 741F8 anti–c–erbB–2 sFv, assembled as either the VH–VL or VL–VH, and expressed alone, with C–terminal cysteine for dimerization, or as fusion proteins with carboxyl–terminal effector domains, including interleukin–2, the B domain of staphylococcal protein A, the S–peptide of ribonuclease S, or hexa–histidine metal chelate peptide. Constructs were expressed and secreted transiently in 293 cells and stably in CHO or Sp2/0 cell lines, the latter yielding up to 10 mg per liter. Single–chain constructs of MOPC 315 myeloma and 26–10 monoclonal antibodies were also expressed, as were hybrids comprising unrelated VH and VL regions. Our results suggest that mammalian expression is a practical and valuable complement to the bacterial expression of single–chain antibodies.

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Author notes

  1. L. L. Houston

    Present address: Prizm Pharmaceuticals, Inc., 10655 Sorrento Valley Road, Suite 200, San Diego, CA, 92101

  2. Hermann Oppermann: Corresponding author.

Authors and Affiliations

  1. Creative BioMolecules, Inc., 45 South Street, Hopkinton, MA, 01748

    Haimanti Dorai, John E. McCartney, Robert M. Hudziak, Mei-Sheng Tai, James S. Huston & Hermann Oppermann

  2. Chiron Corporation, 4560 Horton Street, Emeryville, CA, 94608

    Axel A. Laminet & L. L. Houston

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  1. Haimanti Dorai
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Dorai, H., McCartney, J., Hudziak, R. et al. Mammalian Cell Expression of Single–Chain Fv (sFv) Antibody Proteins and Their C–terminal Fusions with Interleukin–2 and Other Effector Domains. Nat Biotechnol 12, 890–897 (1994). https://doi.org/10.1038/nbt0994-890

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