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Synthetic libraries of shark vNAR domains with different cysteine numbers within the CDR3

The variable domain of New Antigen Receptors (vNAR) from sharks, present special characteristics in comparison to the conventional antibody molecules such as: small size (12–15 kDa), thermal and chemical stability and great tissue penetration, that makes them a good alternative source as therapeutic...

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Autores principales: Cabanillas-Bernal, Olivia, Dueñas, Salvador, Ayala-Avila, Marta, Rucavado, Alexandra, Escalante, Teresa, Licea-Navarro, Alexei F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6576789/
https://www.ncbi.nlm.nih.gov/pubmed/31206542
http://dx.doi.org/10.1371/journal.pone.0213394
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author Cabanillas-Bernal, Olivia
Dueñas, Salvador
Ayala-Avila, Marta
Rucavado, Alexandra
Escalante, Teresa
Licea-Navarro, Alexei F.
author_facet Cabanillas-Bernal, Olivia
Dueñas, Salvador
Ayala-Avila, Marta
Rucavado, Alexandra
Escalante, Teresa
Licea-Navarro, Alexei F.
author_sort Cabanillas-Bernal, Olivia
collection PubMed
description The variable domain of New Antigen Receptors (vNAR) from sharks, present special characteristics in comparison to the conventional antibody molecules such as: small size (12–15 kDa), thermal and chemical stability and great tissue penetration, that makes them a good alternative source as therapeutic or diagnostic agents. Therefore, it is essential to improve techniques used for the development and selection of vNAR antibodies that recognize distinct antigens. The development of synthetic antibody libraries offers a fast option for the generation of antibodies with the desired characteristics. In this work three synthetic antibody libraries were constructed; without cysteines (Cys), with one Cys and with two Cys residues within its CDR3, with the objective of determining whether the presence or absence of Cys in the CDR3 favors the isolation of vNAR clones from a synthetic library. The libraries were validated selecting against six mammalian proteins. At least one vNAR was found for each of the antigens, and a clone coming from the library without Cys in the CDR3 was selected with all the antigens. In vitro angiogenesis assay with the isolated anti-VEGF antibodies, suggest that these vNARs are capable of inhibiting in vitro angiogenesis. In silico analysis of anti-VEGF antibodies showed that vNARs from synthetic libraries could rival antibodies with affinity maturation by in silico modeling.
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spelling pubmed-65767892019-06-28 Synthetic libraries of shark vNAR domains with different cysteine numbers within the CDR3 Cabanillas-Bernal, Olivia Dueñas, Salvador Ayala-Avila, Marta Rucavado, Alexandra Escalante, Teresa Licea-Navarro, Alexei F. PLoS One Research Article The variable domain of New Antigen Receptors (vNAR) from sharks, present special characteristics in comparison to the conventional antibody molecules such as: small size (12–15 kDa), thermal and chemical stability and great tissue penetration, that makes them a good alternative source as therapeutic or diagnostic agents. Therefore, it is essential to improve techniques used for the development and selection of vNAR antibodies that recognize distinct antigens. The development of synthetic antibody libraries offers a fast option for the generation of antibodies with the desired characteristics. In this work three synthetic antibody libraries were constructed; without cysteines (Cys), with one Cys and with two Cys residues within its CDR3, with the objective of determining whether the presence or absence of Cys in the CDR3 favors the isolation of vNAR clones from a synthetic library. The libraries were validated selecting against six mammalian proteins. At least one vNAR was found for each of the antigens, and a clone coming from the library without Cys in the CDR3 was selected with all the antigens. In vitro angiogenesis assay with the isolated anti-VEGF antibodies, suggest that these vNARs are capable of inhibiting in vitro angiogenesis. In silico analysis of anti-VEGF antibodies showed that vNARs from synthetic libraries could rival antibodies with affinity maturation by in silico modeling. Public Library of Science 2019-06-17 /pmc/articles/PMC6576789/ /pubmed/31206542 http://dx.doi.org/10.1371/journal.pone.0213394 Text en © 2019 Cabanillas-Bernal et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Cabanillas-Bernal, Olivia
Dueñas, Salvador
Ayala-Avila, Marta
Rucavado, Alexandra
Escalante, Teresa
Licea-Navarro, Alexei F.
Synthetic libraries of shark vNAR domains with different cysteine numbers within the CDR3
title Synthetic libraries of shark vNAR domains with different cysteine numbers within the CDR3
title_full Synthetic libraries of shark vNAR domains with different cysteine numbers within the CDR3
title_fullStr Synthetic libraries of shark vNAR domains with different cysteine numbers within the CDR3
title_full_unstemmed Synthetic libraries of shark vNAR domains with different cysteine numbers within the CDR3
title_short Synthetic libraries of shark vNAR domains with different cysteine numbers within the CDR3
title_sort synthetic libraries of shark vnar domains with different cysteine numbers within the cdr3
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6576789/
https://www.ncbi.nlm.nih.gov/pubmed/31206542
http://dx.doi.org/10.1371/journal.pone.0213394
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