Mammalian Cell Expression of Single–Chain Fv (sFv) Antibody Proteins and Their C–terminal Fusions with Interleukin–2 and Other Effector Domains
| Autor: | James S. Huston, Haimanti Dorai, Mei-Sheng Tai, Hermann Oppermann, Axel A. Laminet, John E. McCartney, L. L. Houston, Robert M. Hudziak |
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| Rok vydání: | 1994 |
| Předmět: |
Macromolecular Substances
Receptor ErbB-2 medicine.drug_class Recombinant Fusion Proteins Blotting Western Molecular Sequence Data Restriction Mapping Biomedical Engineering Gene Expression Bioengineering CHO Cells Transfection Monoclonal antibody Polymerase Chain Reaction Applied Microbiology and Biotechnology Chromatography Affinity Cell Line Cricetinae Proto-Oncogene Proteins Gene expression Tumor Cells Cultured medicine Animals Humans Amino Acid Sequence Immunoglobulin Fragments DNA Primers Base Sequence biology Effector HEK 293 cells Antibodies Monoclonal Fusion protein Molecular biology Recombinant Proteins ErbB Receptors Cell culture biology.protein Interleukin-2 Molecular Medicine Antibody Plasmacytoma Plasmids Biotechnology |
| Zdroj: | Nature Biotechnology. 12:890-897 |
| ISSN: | 1546-1696 1087-0156 |
| DOI: | 10.1038/nbt0994-890 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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