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Enhancing Stability of Camelid and Shark Single Domain Antibodies: An Overview

Single domain antibodies (sdAbs) are gaining a reputation as superior recognition elements as they combine the advantages of the specificity and affinity found in conventional antibodies with high stability and solubility. Melting temperatures (T(m)s) of sdAbs cover a wide range from below 50 to ove...

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Autores principales: Goldman, Ellen R., Liu, Jinny L., Zabetakis, Dan, Anderson, George P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5524736/
https://www.ncbi.nlm.nih.gov/pubmed/28791022
http://dx.doi.org/10.3389/fimmu.2017.00865
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author Goldman, Ellen R.
Liu, Jinny L.
Zabetakis, Dan
Anderson, George P.
author_facet Goldman, Ellen R.
Liu, Jinny L.
Zabetakis, Dan
Anderson, George P.
author_sort Goldman, Ellen R.
collection PubMed
description Single domain antibodies (sdAbs) are gaining a reputation as superior recognition elements as they combine the advantages of the specificity and affinity found in conventional antibodies with high stability and solubility. Melting temperatures (T(m)s) of sdAbs cover a wide range from below 50 to over 80°C. Many sdAbs have been engineered to increase their T(m), making them stable until exposed to extreme temperatures. SdAbs derived from the variable heavy chains of camelid and shark heavy chain-only antibodies are termed VHH and VNAR, respectively, and generally exhibit some ability to refold and bind antigen after heat denaturation. This ability to refold varies from 0 to 100% and is a property dependent on both intrinsic factors of the sdAb and extrinsic conditions such as the sample buffer ionic strength, pH, and sdAb concentration. SdAbs have also been engineered to increase their solubility and refolding ability, which enable them to function even after exposure to temperatures that exceed their melting point. In addition, efforts to improve their stability at extreme pH and in the presence of chemical denaturants or proteases have been undertaken. Multiple routes have been employed to engineer sdAbs with these enhanced stabilities. The methods utilized to achieve these goals include grafting complementarity-determining regions onto stable frameworks, introduction of non-canonical disulfide bonds, random mutagenesis combined with stringent selection, point mutations such as inclusion of negative charges, and genetic fusions. Increases of up to 20°C have been realized, pushing the T(m) of some sdAbs to over 90°C. Herein, we present an overview of the work done to stabilize sdAbs derived from camelids and sharks. Utilizing these various strategies sdAbs have been stabilized without significantly compromising their affinity, thereby providing superior reagents for detection, diagnostic, and therapeutic applications.
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spelling pubmed-55247362017-08-08 Enhancing Stability of Camelid and Shark Single Domain Antibodies: An Overview Goldman, Ellen R. Liu, Jinny L. Zabetakis, Dan Anderson, George P. Front Immunol Immunology Single domain antibodies (sdAbs) are gaining a reputation as superior recognition elements as they combine the advantages of the specificity and affinity found in conventional antibodies with high stability and solubility. Melting temperatures (T(m)s) of sdAbs cover a wide range from below 50 to over 80°C. Many sdAbs have been engineered to increase their T(m), making them stable until exposed to extreme temperatures. SdAbs derived from the variable heavy chains of camelid and shark heavy chain-only antibodies are termed VHH and VNAR, respectively, and generally exhibit some ability to refold and bind antigen after heat denaturation. This ability to refold varies from 0 to 100% and is a property dependent on both intrinsic factors of the sdAb and extrinsic conditions such as the sample buffer ionic strength, pH, and sdAb concentration. SdAbs have also been engineered to increase their solubility and refolding ability, which enable them to function even after exposure to temperatures that exceed their melting point. In addition, efforts to improve their stability at extreme pH and in the presence of chemical denaturants or proteases have been undertaken. Multiple routes have been employed to engineer sdAbs with these enhanced stabilities. The methods utilized to achieve these goals include grafting complementarity-determining regions onto stable frameworks, introduction of non-canonical disulfide bonds, random mutagenesis combined with stringent selection, point mutations such as inclusion of negative charges, and genetic fusions. Increases of up to 20°C have been realized, pushing the T(m) of some sdAbs to over 90°C. Herein, we present an overview of the work done to stabilize sdAbs derived from camelids and sharks. Utilizing these various strategies sdAbs have been stabilized without significantly compromising their affinity, thereby providing superior reagents for detection, diagnostic, and therapeutic applications. Frontiers Media S.A. 2017-07-25 /pmc/articles/PMC5524736/ /pubmed/28791022 http://dx.doi.org/10.3389/fimmu.2017.00865 Text en Copyright © 2017 Goldman, Liu, Zabetakis and Anderson. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Goldman, Ellen R.
Liu, Jinny L.
Zabetakis, Dan
Anderson, George P.
Enhancing Stability of Camelid and Shark Single Domain Antibodies: An Overview
title Enhancing Stability of Camelid and Shark Single Domain Antibodies: An Overview
title_full Enhancing Stability of Camelid and Shark Single Domain Antibodies: An Overview
title_fullStr Enhancing Stability of Camelid and Shark Single Domain Antibodies: An Overview
title_full_unstemmed Enhancing Stability of Camelid and Shark Single Domain Antibodies: An Overview
title_short Enhancing Stability of Camelid and Shark Single Domain Antibodies: An Overview
title_sort enhancing stability of camelid and shark single domain antibodies: an overview
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5524736/
https://www.ncbi.nlm.nih.gov/pubmed/28791022
http://dx.doi.org/10.3389/fimmu.2017.00865
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