Application of yeast surface display for identification of marine virus specific antibody

Abstract

Marine viruses are the most numerous biological entities in the oceans and probably infect all living things from bacteria to mammal. However, there are few researches on marine viruses to isolate and detect them because of the high mutation rates and variation of viral genomes. We are currently developing new experiment system to identify and detect various marine viruses by using yeast surface display followed by scFv display. Yeast surface display (YSD) is a powerful method for isolating and engineering proteins to increase their affinity, specificity and stability in many different areas. Recently YSD has been introduced to identify and screen a certain antibody to a specific antigen protein. Yeast has eukaryotic characteristics in protein biosynthesis so that antigen protein seems to be similar 3D structures and post translational modification. In this study we expressed the capsid proteins of two marine viruses (RSIV: Red sea bream iridovirus and VNNV: Viral Nervous Necrosis virus) in YSD. Viral capsid genes were synthesized based on sequence data base and subcloned into yeast expression vector. And then these vectors were introduced into EBY100 yeast cells. FACS analysis showed that RSIV and VNNV capsid proteins were expressed good enough to isolate anti RSIV- and VNNV-scFv respectively from scFv phage display library which were prepared in our previous studies. Taken together, YSD and scFc phage display can be a powerfull tool to isolate marine virus specific antibody.