Abstract

Nanobodies are recombinant antigen-specific single domain antibodies derived from the heavy chain–only subset of camelid immunoglobulins. These antibodies, also known as VHHs, hold immense potential in biomedical research due to their small size, high affinity, and stability. 

Raymond Owens' group at the Rosalind Franklin Institute, has been at the forefront of nanobody research. Their work involves generating nanobodies against a diverse array of antigens for applications spanning cell and structural biology. Leveraging phage display libraries constructed from the peripheral blood cells of immunized llamas and alpacas, they have successfully generated nanobodies to over 75 different antigens since 2019. These nanobodies have found utility as structural chaperones, diagnostics, and even anti-viral therapeutics for diseases like SARS-CoV-2.

In this webinar, we present a standardized process for nanobody discovery from Llama, covering immunization, library building, panning, and small-scale expression for binding clone prioritization, as well as cloning strategies to functionalize a subset of nanobodies for further downstream applications such as imaging and purification.

Highlights: 

1. A standardized workflow for nanobody discovery from llama immunization to ligation-independent cloning for VHH domain library construction and small-scale expression screening.

2. Introduction of two suites of vectors with generic VHH cloning primers for nanobody functionalization in bacterial or mammalian cells, enhancing their adaptability for various applications in imaging and purification.