Transgenic Research

, Volume 19, Issue 4, pp 575–586

High-level expression of Camelid nanobodies in Nicotiana benthamiana

Original Paper

DOI: 10.1007/s11248-009-9338-0

Cite this article as:
Teh, YH.A. & Kavanagh, T.A. Transgenic Res (2010) 19: 575. doi:10.1007/s11248-009-9338-0

Abstract

Nanobodies (or VHHs) are single-domain antigen-binding fragments derived from Camelid heavy chain-only antibodies. Their small size, monomeric behaviour, high stability and solubility, and ability to bind epitopes not accessible to conventional antibodies make them especially suitable for many therapeutic and biotechnological applications. Here we describe high-level expression, in Nicotiana benthamiana, of three versions of an anti-hen egg white lysozyme (HEWL) nanobody which include the original VHH from an immunized library (cAbLys3), a codon-optimized derivative, and a codon-optimized hybrid nanobody comprising the CDRs of cAbLys3 grafted onto an alternative ‘universal’ nanobody framework. His6- and StrepII-tagged derivatives of each nanobody were targeted for accumulation in the cytoplasm, chloroplast and apoplast using different pre-sequences. When targeted to the apoplast, intact functional nanobodies accumulated at an exceptionally high level (up to 30% total leaf protein), demonstrating the great potential of plants as a nanobody production system.

Keywords

Camelid VHH Nanobody Anti-lysozyme Nicotiana benthamiana 

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  1. 1.Plant Molecular Genetics Laboratory, Smurfit Institute of GeneticsTrinity CollegeDublin 2Ireland