Nanobody

The Nanobody facility provides members of the LMB with a service for isolating single-chain camelid nanobodies that bind to proteins and complexes of their choice.

The facility advises on all aspects of the procedure from target protein stabilisation and tagging through to the expression and purification of nanobodies.

Researchers provide purified protein suitable for immunisation of alpacas and the facility then organises the injections, blood collection, isolation of peripheral blood mononuclear cells (PBMCs), construction of phage libraries, panning against the target protein and isolation of enriched nanobodies, using either traditional methods such as ELISA or next-generation sequencing (NGS). The facility can also express and purify nanobodies to determine their binding affinity to targets using bilayer interferometry, surface plasmon resonance or fluorescence polarisation spectroscopy.

Nanobodies have been raised successfully to single-domain proteins, large complexes, integral membrane proteins and protein filaments, with affinities typically in the range of double-digit nanomolar to high picomolar.