From llama to nanobody: a streamlined workflow for the generation of functionalised VHHs

Eyssen, L. E. -A., Ramadurai, S., Abdelkarim, S., Buckle, I., Cornish, K., Lin, H., Jones, A. K., Stephens, G. J. ORCID: https://orcid.org/0000-0002-8966-4238 and Owens, R. J. (2024) From llama to nanobody: a streamlined workflow for the generation of functionalised VHHs. Bio-Protocol, 14 (6). e4962. ISSN 2331-8325 doi: 10.21769/BioProtoc.4962

Abstract/Summary

Nanobodies are recombinant antigen-specific single domain antibodies (VHHs) derived from the heavy chain–only subset of camelid immunoglobulins. Their small molecular size, facile expression, high affinity, and stability have combined to make them unique targeting reagents with numerous applications in the biomedical sciences. From our work in producing nanobodies to over sixty different proteins, we present a standardised workflow for nanobody discovery from llama immunisation, library building, panning, and small-scale expression for prioritisation of binding clones. In addition, we introduce our suites of mammalian and bacterial vectors, which can be used to functionalise selected nanobodies for various applications such as in imaging and purification.

Item Type	Article
URI	https://reading-clone.eprints-hosting.org/id/eprint/115863
Identification Number/DOI	10.21769/BioProtoc.4962
Refereed	Yes
Divisions	Interdisciplinary Research Centres (IDRCs) > Centre for Integrative Neuroscience and Neurodynamics (CINN) Life Sciences > School of Chemistry, Food and Pharmacy > School of Pharmacy > Division of Pharmacology
Publisher	Bio-protocol LLC
Download/View statistics	View download statistics for this item