SELECTION OF AN SCFV PHAGE ANTIBODY THAT RECOGNIZES BLUETONGUE VIRUS FROM A LARGE SYNTHETIC LIBRARY AND ITS USE IN ELISAS TO DETECT VIRAL-ANTIGEN AND ANTIBODIES
W. Vanwyngaardt et Dh. Duplessis, SELECTION OF AN SCFV PHAGE ANTIBODY THAT RECOGNIZES BLUETONGUE VIRUS FROM A LARGE SYNTHETIC LIBRARY AND ITS USE IN ELISAS TO DETECT VIRAL-ANTIGEN AND ANTIBODIES, Onderstepoort journal of veterinary research, 65(2), 1998, pp. 125-131
A filamentous phage library displaying a vast repertoire of synthetic
single chain fragment variable (scFv) antibody fragments was subjected
to affinity selection on purified bluetongue virus (BTV) particles. A
fter four rounds of selection and amplification, 73 out of a total of
90 fusion phage clones tested were found to bind to purified BTV in EL
ISA. One of these, the clone producing the highest ELISA signal, was s
elected for an investigation of its potential as an immunodiagnostic r
eagent. The binding of this phage antibody (designated A12) could be i
nhibited by free virus and by antibodies in immune serum. Inhibition w
ith antibodies in guinea-pig sera suggested that it recognized an anti
genic region on BTV that was similar on at least 10 different BTV sero
types. A sandwich ELISA utilizing antibody A12 was capable of detectin
g approximately 60 ng of purified BTV.