Cost per project estimate: €5500
-immunisation up to 3 proteins
- screening method developed for specificity
- Nanobody repertoire amplified by RT-PCR
- cloned into phage display libraries
- Nanobodies selected
Description of service
Nanobodies are compact single Ig domains that bind target proteins with high affinity while reducing their coinformational heterogeneity, thereby stabilising multi-protein complexes. Their properties mean that nanobodies are versatile tools for structural biology.
Scientific relevance: Nanobody production has several applications relevant to vaccine design:
Nanobody/protein complexes may be more stable than individual proteins, favouring crystallisation
for X-ray analysis (including intrinsically disordered proteins)
The CDR3 loop of nanobodies can extend into small cavities on antigenic surfaces which otherwise are
difficult to measure or modulate, and nanobodies do not compete with classical antibodies that bind
the same antigen. Some are in development as therapeutic agents.
Nanobody reagents can capture transient protein conformations
Nanobodies can be immuno-labelled for super-resolution imaging, spin-labelled for EPR spectroscopy
or heavy metal labelled.
Small size can be used in NMR methods, can act as NMR invisible probes for dynamic studies of protein
(longer if re-immunisation is required)
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Example publications of nanobody use for structural analysis:
J Struct Biol. 2011 Apr; 174(1): 124–136. doi: 10.1016/j.jsb.2010.10.007 ‘STRUCTURES OF A KEY INTERACTION PROTEIN FROM THE TRYPANOSOMA BRUCEI EDITOSOME IN COMPLEX WITH SINGLE DOMAIN ANTIBODIES.’ Meiting Wu, Young-jun Park, Els Pardon, Stewart Turley, Andrew Hayhurst, Junpeng Deng, Jan Steyaert, and Wim G. J. Hol.
Vaccines (Basel). 2019 Sep; 7(3): 77. doi: 10.3390/vaccines7030077 ‘Nanobodies that Neutralize HIV.’ Robin A. Weiss and C. Theo Verrips.
Sample Requirements - input of users
2mg purified protein for immunisation (or cells expressing protein, or genetic vaccination)
Good assay for selecting nanobodies with appropriate properties