The purpose of this project was to isolate recombinant antibodies for
diagnosis of very virulent infectious bursal disease virus (vvIBDV) in
fixed tissues. Phage-displayed recombinant antibodies, comprised of the
single chain variable fragment (scFv), were investigated. A previously
made recombinant antibody library generated against vvIBDV was selected
and screened for recombinant antibodies that reacted against vvIBDV in
ELISA. A new library was constructed fom chickens immunized with fixed
vvIBDV and also screened for recombinant antibodies against vvIBDV. Also,
a previously identified recombinant antibody, known to react well with
vvIBDV, was used to replace either the Vh or VI gene with corresponding
fragments from a new library. The Vh and VI antibody genes were initially
amplified effectively by PCR. No new recombinant antibody clones were
isolated from the libraries generated against vvIBDV. However, exchanging
the Vh or VI genes from a known recombinant antibody with genes from a
new library showed that the heavy chain was essential when binding to
vvBDV. Light chains could be exchanged without loss of activity, but when
heavy chains were exchanged, all activity was lost. The light chains were
found to create new binding properties when combined with the essential
heavy chain. The recombinant antibody clones were sequenced, analyszd and
characterize.