Human single-chain variable fragment antibody inhibits macrophage migration inhibitory factor tautomerase activity
| Autor: | Ornnuthchar Poungpair, Duangporn Ungsupravate, Kunan Bangphoomi, Pa-thai Yenchitsomanus, Wanpen Chaicumpa, Mayuri Tarasuk |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Phage display
medicine.medical_treatment animal diseases Anti-Inflammatory Agents chemical and pharmacologic phenomena Enzyme-Linked Immunosorbent Assay Biology law.invention Immune system Western blot law Catalytic Domain Cell Line Tumor Genetics medicine otorhinolaryngologic diseases Humans Macrophage Migration-Inhibitory Factors Leukemia medicine.diagnostic_test Mimotope General Medicine Articles Molecular biology Immunity Innate inflammatory disease Intramolecular Oxidoreductases Cytokine inflammation tautomerase activity Recombinant DNA biology.protein macrophage migration inhibitory factor Macrophage migration inhibitory factor Antibody human single-chain variable fragment antibody Protein Binding Single-Chain Antibodies |
| Zdroj: | International Journal of Molecular Medicine |
| ISSN: | 1791-244X 1107-3756 |
| Popis: | Macrophage migration inhibitory factor (MIF) is a pro-inflammatory cytokine, secreted from a variety of immune cells, that regulates innate and adaptive immune responses. Elevation of MIF levels in plasma correlates with the severity of inflammatory diseases in humans. Inhibition of MIF or its tautomerase activity ameliorates disease severity by reducing inflammatory responses. In this study, the human single-chain variable fragment (HuScFv) antibody specific to MIF was selected from the human antibody phage display library by using purified recombinant full-length human MIF (rMIF) as the target antigen. Monoclonal HuScFv was produced from phage-transformed bacteria and tested for their binding activities to rMIF by indirect enzyme-linked immunosorbent assay as well as to native MIF by western blot analysis and immunofluorescence assay. The HuScFv with highest binding signal to rMIF also inhibited the tautomerase activities of both rMIF and native MIF in human monoblastic leukemia (U937) cells in a dose-dependent manner. Mimotope searching and molecular docking concordantly demonstrated that the HuScFv interacted with Lys32 and Ile64 in the MIF tautomerase active site. To the best of our knowledge, this is the first study to focus on MIF-specific fully-human antibody fragment with a tautomerase-inhibitory effect that has potential to be developed as anti-inflammatory biomolecules for human use. |
| Databáze: | OpenAIRE |
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