Generation of a murine single chain Fv (scFv) antibody specific for cucumber mosaic virus (CMV) using a phage display library.
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(H)) gene belonged to the VH3 subgroup and the kappa light chain variable region (V kappa) came from the Vkappa4 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.
Chae
J S
JS
Division of Food Science & Biotechnology, College of Agriculture & Life Sciences, Kangwon National University, Chunchon, Korea.
Choi
J K
JK
Lim
H T
HT
Cha
S H
SH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000737:chemistry / Q000235:genetics*; D000918:Antibody Specificity; D001483:Base Sequence; D018553:Cucumis sativus / Q000235:genetics / Q000821:virology; D017799:Cucumovirus / Q000235:genetics / Q000276:immunology*; D018076:DNA, Complementary; D005820:Genetic Testing; D006825:Hybridomas; D007135:Immunoglobulin Variable Region / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D030821:Plants, Genetically Modified; D011485:Protein Binding / Q000276:immunology; D057127:Single-Chain Antibodies; D013154:Spleen / Q000166:cytology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D018076:DNA, Complementary; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D057127:Single-Chain Antibodies; C114805:scFv fragment SW1