Generation of chicken single chain antibody variable fragments (scFv) that differentiate and neutralize infectious bursal disease virus (IBDV)

摘要: Phage-displayed recombinant antibody libraries derived from splenic mRNA of chickens immunized with an Australian strain infectious bursal disease virus (IBDV) were constructed as single chain variable fragments (scFv) by either overlap extension polymerase reaction (PCR) or sequential ligation the individual heavy (VH) and light (VL) gene segments. Sequential cloning VH VL genes into a newly pCANTAB-link vector containing synthetic linker sequence (Gly4Ser)3 was more efficient than PCR, increasing library size 500 fold. Eighteen IBDV specific antibodies unique scFv sequences identified after panning against immunizing antigen. Eight clones contained identical but genes. In ELISA analysis using panel overseas strains, one able to detect all whilst 3 others could discriminate between classical variant strains field vaccine strains. addition, some scFvs showed significant neutralization titres in vitro. This report shows that generation chicken vitro means has considerable potential for producing diverse specificity neutralizing capacity.