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

Author(s): Ema Romao , Francisco Morales-Yanez , Yaozhong Hu , Maxine Crauwels , Pieter De Pauw , Gholamreza Ghassanzadeh Hassanzadeh , Nick Devoogdt , Chloe Ackaert , Cecile Vincke , Serge Muyldermans .

Journal Name: Current Pharmaceutical Design

Volume 22 , Issue 43 , 2016

Abstract:

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.

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Article Details

VOLUME: 22
ISSUE: 43
Year: 2016
Page: [6500 - 6518]
Pages: 19
DOI: 10.2174/1381612822666160923114417
Price: $58

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