Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans

Sy Jye Leu, Yu Ching Lee, Chi Hsin Lee, Po Yen Liao, Chen Wei Chiang, Chieh Ming Yang, Ching Hua Su, Tsong Yih Ou, Ko Jiunn Liu, Hsiu Jung Lo, Bor Yu Tsai, Yi Yuan Yang

Research output: Contribution to journalArticle

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.

Original languageEnglish
JournalInternational journal of molecular sciences
Volume21
Issue number8
DOIs
Publication statusPublished - Apr 21 2020

Keywords

  • alpha–enolase
  • C. albicans
  • phage display technology
  • single chain variable fragment

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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  • Cite this

    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). https://doi.org/10.3390/ijms21082903