High-level expression of <Emphasis Type="Italic">Camelid</Emphasis> nanobodies in <Emphasis Type="Italic">Nicotiana benthamiana</Emphasis> |
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Authors: | Yi-Hui Audrey Teh Tony A Kavanagh |
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Institution: | (1) Plant Molecular Genetics Laboratory, Smurfit Institute of Genetics, Trinity College, Dublin 2, Ireland; |
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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. |
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Keywords: | |
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