Identification of Useful Nanobodies by Phage Display of Immune Single Domain Libraries Derived from Camelid Heavy Chain Antibodies

Romao, Ema; Morales Yánez, Francisco Javier; Hu, Yaozhong; Crauwels, Maxine; De Pauw, Pieter; Ghassanzadeh Hassanzadeh, Gholamreza; Devoogdt, Nick; Ackaert, Chloe; Vincke, Cecile; Muyldermans, Serge

doi:10.2174/1381612822666160923114417

Article (Scientific journals)

Identification of Useful Nanobodies by Phage Display of Immune Single Domain Libraries Derived from Camelid Heavy Chain Antibodies

Romao, Ema; Morales Yánez, Francisco Javier; Hu, Yaozhong et al.

2016 • In Current Pharmaceutical Design

Peer Reviewed verified by ORBi

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https://hdl.handle.net/2268/267106

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10.2174/1381612822666160923114417

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27669966

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Abstract :

[en] Background: The discovery of functional heavy chain-only antibodies devoid of light chains in sera of camelids and sharks in the early nineties provided access to the generation of minimal-sized, single-domain, in vivo affinity-matured, recombinant antigenbinding fragments, also known as Nanobodies. Methods: Recombinant DNA technology and adaptation of phage display vectors form the basis to construct large naïve, synthetic or medium sized immune libraries from where multiple Nanobodies have been retrieved. Alternative selection methods (i.e. bacterial display, bacterial two-hybrid, Cis-display and ribosome display) have also been developed to identify Nanobodies. The antigen affinity, stability, expression yields and structural details of the Nanobodies have been determined by standard technology. Nanobodies were subsequently engineered for higher stability and affinity, to have a sequence closer to that of human immunoglobulin domains, or to add designed effector functions. Results: Antigen specific Nanobodies recognizing with high affinity their cognate antigen were retrieved from various libraries. High expression yields are obtained from microorganisms, even when expressed in the cytoplasm. The purified Nanobodies are shown to possess beneficial biochemical and biophysical properties. The crystal structure of Nanobody::antigen complexes reveal the preference of Nanobodies for cavities on the antigen surface. Conclusion: Thanks to the properties described above, Nanobodies became a highly valued and versatile tool for biomolecular research. Moreover, numerous diagnostic and therapeutic Nanobody-based applications have been developed in the past decade. Keywords: Cell biology tools; diagnostics; immune libraries; nanobodies; research tools; single domain antibodies; therapeutics.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Romao, Ema; Vrije Universiteit Brussel - VUB > Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.

Morales Yánez, Francisco Javier ; Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines

Hu, Yaozhong; Vrije Universiteit Brussel - VUB > Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.

Crauwels, Maxine; Vrije Universiteit Brussel - VUB > Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.

De Pauw, Pieter; Vrije Universiteit Brussel - VUB > Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.

Ghassanzadeh Hassanzadeh, Gholamreza; Vrije Universiteit Brussel - VUB > Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.

Devoogdt, Nick; Vrije Universiteit Brussel - VUB > Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.

Ackaert, Chloe; Vrije Universiteit Brussel - VUB > Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.

Vincke, Cecile; Vrije Universiteit Brussel - VUB > Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.

Muyldermans, Serge; Vrije Universiteit Brussel - VUB > Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium

Language :

English

Title :

Identification of Useful Nanobodies by Phage Display of Immune Single Domain Libraries Derived from Camelid Heavy Chain Antibodies

Publication date :

2016

Journal title :

Current Pharmaceutical Design

ISSN :

1381-6128

eISSN :

1873-4286

Publisher :

Bentham Science Publishers, United Arab Emirates

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 17 January 2022

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