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



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.



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.



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.
Ema
E
Romao
Francisco
F
Morales-Yanez
Yaozhong
Y
Hu
Maxine
M
Crauwels
Pieter
P
De Pauw
Gholamreza Ghassanzadeh
GG
Hassanzadeh
Nick
N
Devoogdt
Chloe
C
Ackaert
Cecile
C
Vincke
Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.
Serge
S
Muyldermans
D000818:Animals; D001435:Bacteriophages / Q000235:genetics*; D002162:Camelus; D006801:Humans; D019151:Peptide Library; D061905:Single-Domain Antibodies / Q000235:genetics / Q000276:immunology*
D019151:Peptide Library; D061905:Single-Domain Antibodies
Cell biology tools; diagnostics; immune libraries; nanobodies; research tools; single domain antibodies; therapeutics.