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

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...

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Autores principales: Leu, Sy-Jye, Lee, Yu-Ching, Lee, Chi-Hsin, Liao, Po-Yen, Chiang, Chen-Wei, Yang, Chieh-Ming, Su, Ching-Hua, Ou, Tsong-Yih, Liu, Ko-Jiunn, Lo, Hsiu-Jung, Tsai, Bor-Yu, Yang, Yi-Yuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7215377/
https://www.ncbi.nlm.nih.gov/pubmed/32326294
http://dx.doi.org/10.3390/ijms21082903
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author Leu, Sy-Jye
Lee, Yu-Ching
Lee, Chi-Hsin
Liao, Po-Yen
Chiang, Chen-Wei
Yang, Chieh-Ming
Su, Ching-Hua
Ou, Tsong-Yih
Liu, Ko-Jiunn
Lo, Hsiu-Jung
Tsai, Bor-Yu
Yang, Yi-Yuan
author_facet Leu, Sy-Jye
Lee, Yu-Ching
Lee, Chi-Hsin
Liao, Po-Yen
Chiang, Chen-Wei
Yang, Chieh-Ming
Su, Ching-Hua
Ou, Tsong-Yih
Liu, Ko-Jiunn
Lo, Hsiu-Jung
Tsai, Bor-Yu
Yang, Yi-Yuan
author_sort Leu, Sy-Jye
collection PubMed
description 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.
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spelling pubmed-72153772020-05-18 Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans Leu, Sy-Jye Lee, Yu-Ching Lee, Chi-Hsin Liao, Po-Yen Chiang, Chen-Wei Yang, Chieh-Ming Su, Ching-Hua Ou, Tsong-Yih Liu, Ko-Jiunn Lo, Hsiu-Jung Tsai, Bor-Yu Yang, Yi-Yuan Int J Mol Sci Article 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. MDPI 2020-04-21 /pmc/articles/PMC7215377/ /pubmed/32326294 http://dx.doi.org/10.3390/ijms21082903 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Leu, Sy-Jye
Lee, Yu-Ching
Lee, Chi-Hsin
Liao, Po-Yen
Chiang, Chen-Wei
Yang, Chieh-Ming
Su, Ching-Hua
Ou, Tsong-Yih
Liu, Ko-Jiunn
Lo, Hsiu-Jung
Tsai, Bor-Yu
Yang, Yi-Yuan
Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans
title Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans
title_full Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans
title_fullStr Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans
title_full_unstemmed Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans
title_short Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans
title_sort generation and characterization of single chain variable fragment against alpha-enolase of candida albicans
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7215377/
https://www.ncbi.nlm.nih.gov/pubmed/32326294
http://dx.doi.org/10.3390/ijms21082903
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