Nanobody discovery platform: From a Llama immune response to tail | 35194
Journal of Proteomics & Bioinformatics

Journal of Proteomics & Bioinformatics
Open Access

ISSN: 0974-276X

+44 1223 790975

Nanobody discovery platform: From a Llama immune response to tailor-made biological agents

2nd International Conference on Genetic & Protein Engineering

November 14-16, 2016 Atlanta, Georgia, USA

Tomasz Uchanski

Structural Biology Research Center, Belgium

Posters & Accepted Abstracts: J Proteomics Bioinform

Abstract :

For decades, antibodies have played a central role as research tools and more recently also as therapeutic agents. Their impact on research and medicine is still growing. Nanobodies (Nbs) are the small (15 kDa) and stable single-domain antibody fragments harboring the full antigen-binding capacity of the original heavy chain-only antibodies that naturally occur in camelids. Over the last years, the Steyaert lab pioneered the use of Nbs in structural biology. Nbs are encoded by single gene fragments, are easily produced in microorganisms and exhibit a superior stability compared to the derivatives of conventional antibodies like Fabs or scFvs. Because of their compact prolate shape, Nbs expose a convex paratope and have access to cavities or clefts on the surface of proteins often inaccessible to conventional antibodies. We have demonstrated that Nbs are exquisite antibodies that can bind the most difficult targets including membrane proteins, transient multiprotein assemblies, transient conformational states and intrinsically disordered proteins. Our Nanobody discovery platform utilizes routinely phage display approach and more recently, shifting to the yeast display technology. In yeast display system each yeast cell is capable of displaying up to 100,000 copies of one Nb clone. Thereafter each yeast cell can be stained and analyzed for binding of the fluorescently labeled antigen. Fluorescence-Activated Cell Sorting (FACS) allows the fine-resolution separation of cells by measuring a number of distinct optical parameters in parallel. We are presenting our modified yeast display system, where displayed Nbs are fluorescently labeled in the one-step orthogonally staining. Features of selected Nbs can be first easily characterized using the power of FACS, without a need of recloning, expression and purification of each Nb. The Nanobody discovery platform is opened for diverse collaboration partners through Instruct Integrating Biology.

Biography :

In 2013 he obtained master degree in biotechnology at the Jagiellonia University in Krakow, Poland. The master project was focused on developing a novel scFv expression system. He gained new experience in Fab synthetic library generation and phage display technology during one-year internship program at The University of Chicago, USA. In October 2014 he became a PhD student of Structural Biology Brussels, Belgium. From that point he has had the opportunity to participate in a number of research topics as a Jan Steyaert’s laboratory member. Most of them are focused on protein engineering and different methods of protein display of yeast, bacterial or phage surface. Recently he has been focusing on developing a new yeast display technology for Nanobody discovery platform.