Development of VHH nanobod􀁜 libraries against norovirus G1.I and GII.4

GARAICOECHEA, LORENA; AGUILAR, ANDREA; BOK, MARINA; GREEM, KIM; BOK, KARIN; PARREÑO, VIVIANA

Madison

Congreso; 31 American society for virology annual meeting; 2012

ASV AAM

Development of VHH nanobod􀁜 libraries against norovirus G1.I and GII.4 Lorena Garaicoechea1, Andrea Aguilar1, Marina Bok1, Kim Green2, Karin Bok2, Viviana Parre􀁸o1. 1Instituto de Virología, INTA. Buenos Aires, Argentina., 2Laboratory of Infectious Diseases, NIAID, National Institutes of Health, Bethesda, MD 􀀃􀀃 Norovirus (NoV) is the major cause of epidemic gastroenteritis in humans and the second most important cause of severe diarrhea in children. There is no in vitro cell culture system to isolate and propagate the virus, impairing antigenic studies. Additionally, there are no vaccines or therapeutic drugs for NoV diarrhea. Llama- derived single chain antibody fragments (VHH) are the smallest molecules in nature with antigen binding capacity. The aim of this study was to obtain recombinant monoclonal VHH derived from llama heavy chain antibodies, specific for two distinct NoV genogroups, GI.1 (Norwalk- 1968) and GII.4 (MD-2004) with potential for use in diagnostic assays, treatment, and immunoprophylaxis. Two VHH libraries were developed and four hundred VHH clones were tested by phage ELISA to evaluate specificity to GI.1 and GII.4 genotypes of NoV. Five specific VHH to GI.1, five specific VHH to GII.4 and one cross-reactive VHH were selected and subcloned for further characterization.