A tumor necrosis factor (TNF) receptor-IgG heavy chain chimeric protein as a bivalent antagonist of TNF activity
| Autor: | Bruce Beutler, K. Peppel, David Crawford |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
medicine.drug_class
Recombinant Fusion Proteins medicine.medical_treatment Molecular Sequence Data Immunology Receptors Cell Surface CHO Cells Biology Monoclonal antibody Chromatography Affinity Receptors Tumor Necrosis Factor Thrombin Cricetinae Complementary DNA Tumor Cells Cultured medicine Animals Humans Immunology and Allergy Receptor Base Sequence Tumor Necrosis Factor-alpha Chinese hamster ovary cell Antibodies Monoclonal Articles Molecular biology Fusion protein TNF inhibitor Immunoglobulin G Tumor necrosis factor alpha Immunoglobulin Heavy Chains medicine.drug |
| Zdroj: | The Journal of Experimental Medicine |
| ISSN: | 1540-9538 0022-1007 |
| DOI: | 10.1084/jem.174.6.1483 |
| Popis: | Using a multistep polymerase chain reaction method, we have produced a construct in which a cDNA sequence encoding the extracellular domain of the human 55-kD tumor necrosis factor (TNF) receptor is attached to a sequence encoding the Fc portion and hinge region of a mouse IgG1 heavy chain through an oligomer encoding a thrombin-sensitive peptide linker. This construct was placed downstream from a cytomegalovirus promoter sequence, and expressed in Chinese hamster ovary cells. A secreted protein, capable of binding TNF and inactivating it, was produced by the transfected cells. Molecular characterization revealed that this soluble version of the TNF receptor was dimeric. Moreover, the protein could be quantitatively cleaved by treatment with thrombin. However, the monovalent extracellular domain prepared in this way has a greatly reduced TNF inhibitory activity compared with that of the bivalent inhibitor. Perhaps because of its high affinity for TNF, the chimeric protein is far more effective as a TNF inhibitor than are neutralizing monoclonal antibodies. This molecule may prove very useful as a reagent for the antagonism and assay of TNF and lymphotoxin from diverse species in health and disease, and as a means of deciphering the exact mechanism through which TNF interacts with the 55-kD receptor. |
| Databáze: | OpenAIRE |
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