Mono- and multimeric single chain variable fragments (scFvs), expressed in Pichia pastoris, are capable of neutralizing Bovine Herpesvirus type-1 (BoHV-1) in vitro [Koti et al. 2011. Vaccine 29; Pasman et al., 2012. Clin Vaccine Immunol 19]. The cDNA encoding these scFvs originated from a mouse x cattle hybridoma that secreted bovine IgG1 capable of neutralizing BoHV-1. The molecular modelling of heavy (VH) and light (VL) chain domains predicted that the VL might only have a supportive role in antigen recognition. VH and VL were expressed as single domains (Fd), in P. pastoris with an objective to assess their role in viral recognition and neutralization. The ability to recognize and neutralize BoHV-1 of the monomeric scFv3-18L, VH and VL linked via 18 amino acid linker, was compared to the ability of the individual FdVH and FdVL domains, in an ELISA and in vitro plaque reduction assays. These experiments demonstrated that while FdVH, but not FdVL, recognized BoHV-1 in an ELISA, FdVH did not neutralize BoHV-1. The complexities associated with such antibody functions will be discussed. [Supported by NSERC Canada]
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