PUBLICATION
Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans
- Authors
- Leu, S.J., Lee, Y.C., Lee, C.H., Liao, P.Y., Chiang, C.W., Yang, C.M., Su, C.H., Ou, T.Y., Liu, K.J., Lo, H.J., Tsai, B.Y., Yang, Y.Y.
- ID
- ZDB-PUB-200426-8
- Date
- 2020
- Source
- International Journal of Molecular Sciences 21(8): (Journal)
- Registered Authors
- Keywords
- C. albicans, alpha–enolase, phage display technology, single chain variable fragment
- MeSH Terms
-
- Animals
- Antifungal Agents/pharmacology*
- Candida albicans/drug effects*
- Candida albicans/enzymology*
- Drug Evaluation, Preclinical
- Enzyme Inhibitors/pharmacology*
- Mice
- Phosphopyruvate Hydratase/antagonists & inhibitors*
- Protein Binding
- Single-Chain Antibodies/pharmacology*
- Zebrafish
- PubMed
- 32326294 Full text @ Int. J. Mol. Sci.
Citation
Leu, S.J., Lee, Y.C., Lee, C.H., Liao, P.Y., Chiang, C.W., Yang, C.M., Su, C.H., Ou, T.Y., Liu, K.J., Lo, H.J., Tsai, B.Y., Yang, Y.Y. (2020) Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans. International Journal of Molecular Sciences. 21(8).
Abstract
Candida albicans (C. albicans) is an opportunistic human pathogen responsible for approximately a half of clinical candidemia. The emerging Candida spp. with resistance to azoles is a major challenge in clinic, suggesting an urgent demand for new drugs and therapeutic strategies. Alpha-enolase (Eno1) is a multifunctional protein and represents an important marker for invasive candidiasis. Thus, C. albicans Eno1 (CaEno1) is believed to be an important target for the development of therapeutic agents and antibody drugs. Recombinant CaEno1 (rCaEno1) was first used to immunize chickens. Subsequently, we used phage display technology to construct two single chain variable fragment (scFv) antibody libraries. A novel biopanning procedure was carried out to screen anti-rCaEno1 scFv antibodies, whose specificities were further characterized. The polyclonal IgY antibodies showed binding to rCaEno1 and native CaEno1. A dominant scFv (CaS1) and its properties were further characterized. CaS1 attenuated the growth of C. albicans and inhibited the binding of CaEno1 to plasminogen. Animal studies showed that CaS1 prolonged the survival rate of mice and zebrafish with candidiasis. The fungal burden in kidney and spleen, as well as level of inflammatory cytokines were significantly reduced in CaS1-treated mice. These results suggest CaS1 has potential of being immunotherapeutic drug against C. albicans infections.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping