Nucleic Acids Research, Vol 26, Issue 7 1718-1723, Copyright © 1998 by Oxford University Press
W Ernst, R Grabherr, D Wegner, N Borth, A Grassauer and H Katinger
The baculovirus expression system was utilized to serve as a tool for
ligand selection, demonstrating the applicability of the system to the
generation and screening of eukaryotic expression libraries. The HIV-1-
gp41 epitope 'ELDKWA', specific for the neutralizing human mAb 2F5, was
inserted into the antigenic site B of influenza virus hemagglutinin and
expressed on the surface of baculovirus infected insect cells. In order to
improve the antigenicity of the epitope within the hemagglutinin, and
therefore enhance the specific binding of 2F5, we inserted three
additional, random amino acids adjacent to the epitope. This pool of
hemagglutinin genes was directly cloned into the baculovirus Ac-omega. To
identify distinct proteins displayed on the cellular surface, we developed
a screening protocol to select for specific binding capacity of individual
viral clones. Using fluorescence activated cell sorting (FACS) we isolated
a baculovirus clone displaying the epitope with markedly increased binding
capacity out of a pool of 8000 variants in only one sorting step. Binding
properties of the identified ligand were examined by FACS performing a
competition assay.
ARTICLES
Baculovirus surface display: construction and screening of a eukaryotic epitope library
Insitute of Applied Microbiology, University of Agriculture, Muthgasse 18, A-1190 Vienna, Austria.
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