Generation of a Murine Single Chain Fv (scFv) Antibody Specific for Cucumber Mosaic Virus (CMV) Using a Phage Display Library

초록

With the long-term goal of generating CMV-resistant transgenic plants using antibody genes, a single-chain variable fragment (scFv) antibody that binds to the cucumber mosaic virus was isolated from a scFv phage display library by four rounds of affinity selection with CMV-Mf as an antigen. The scFv has the identical binding specificity to CMV as a monoclonal antibody that is generated by the hybridoma fusion technique, and recognized purified preparations of CMV isolates belonging to either subgroup I or II in immunoblotting. The nucleotide sequences of the recombinant antibody showed that a heavy chain variable region (V<inf>H</inf>) gene belonged to the V<inf>H</inf>3 subgroup and the kappa light chain variable region (V kappa) came from the V<inf>K</inf>4 subgroup. Our results demonstrate that the scFv phage display library, an alternative approach to the traditional hybridoma fusion technique, has a potential applicability in the study of plant virus and plant pathology.

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