Publication: Human single-chain variable fragment antibody inhibits macrophage migration inhibitory factor tautomerase activity
Issued Date
2014-03-01
Resource Type
ISSN
1791244X
11073756
11073756
Other identifier(s)
2-s2.0-84893836912
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
International Journal of Molecular Medicine. Vol.33, No.3 (2014), 515-522
Suggested Citation
Mayuri Tarasuk, Ornnuthchar Poungpair, Duangporn Ungsupravate, Kunan Bangphoomi, Wanpen Chaicumpa, Pa Thai Yenchitsomanus Human single-chain variable fragment antibody inhibits macrophage migration inhibitory factor tautomerase activity. International Journal of Molecular Medicine. Vol.33, No.3 (2014), 515-522. doi:10.3892/ijmm.2014.1622 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/33293
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Human single-chain variable fragment antibody inhibits macrophage migration inhibitory factor tautomerase activity
Other Contributor(s)
Abstract
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.