Characterization of folding-sensitive nanobodies as tools to study the expression and quality of protein particle immunogens.

VANINA ALZOGARAY (1RA AUTORIA COMPARTIDA); MARIELA URRUTIA (1RA AUTORIA COMPARTIDA); PAULA BERGUER; ANDRÉS ROSSI; ROMINA PARDO; VANESA ZYLBERMAN; HERNÁN BONOMI; FERNANDO A GOLDBAUM

JOURNAL OF BIOTECHNOLOGY

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2019

0168-1656

*Alzogaray and Urrutia contributed equally to this work. Vaccination is as one of the most beneficial biopharmaceuticalinterventions against pathogens due to its ability to induce adaptiveimmunity through targeted activation of the immune system. Each vaccineneeds a tailor-made set of tests in order to monitor its qualitythroughout the development and manufacturing. The analysis of theconformational state of protein nanoparticles is one of the key steps invaccine quality control. The enzyme lumazine synthase from Brucella spp.(BLS) acts as a potent oral and systemic immunogen. BLS has been used asa carrier of foreign peptides, protein domains and whole proteins,serving as a versatile platform for vaccine engineering purposes. Here,we show the generation and characterization of four families ofnanobodies (Nbs) which only recognize BLS in its native conformationalstate and that bind to its active site. The present results support theuse of conformation-sensitive Nbs as molecular probes during thedevelopment and production of vaccines based on the BLS platform.Finally, we propose Nbs as useful molecular tools targeting other proteinscaffolds with potential applications in nano-and biotechnology.