OBJECTIVE: To generate phage-displayed anti-idiotypic antibody single chain variable fragments (anti-Id ScFv) MG7 to monoclonal antibody directed against @entity356 so as to lay a foundation for developing anti-Id ScFv vaccine of the @entity5 . METHODS: Balb/c @entity19 were immunized i.p. with purified MG7 monoclonal antibody conjugated with keyhole limpet hemocyanin. mRNA was isolated from the spleens of immunized @entity19 . Heavy and light chain genes (VH and VL) of antibody were amplified separately and assembled into ScFv genes with a specially constructed linker DNA by RT-PCR. The ScFv genes were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into competent @entity526 TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage, which displayed ScFv fragments as a fusion with gene 3 protein on the tips of M13 phage. After four rounds of panning with monoclonal antibody MG7, the MG7-positive clones were selected with the enzyme-linked immunosorbent assay (ELISA) from the enriched phages. The types of the anti-Id ScFv displayed on the selected phage clones were primarily identified by competition ELISA. RESULTS: The VH, VL and ScFv DNAs were about 340, 320 and 750 bp respectively. Twenty-four MG7-positive clones were selected from 40 enriched phage clones. Five of the 24 clones displayed beta or gamma type anti-Id ScFv. CONCLUSION: The anti-Id ScFv frangments to MG7 monoclonal antibody can be successfully selected by recombinant phage antibody technique, which paves a way for the study of prevention and cure of @entity356 using anti-Id ScFv.

[Production of phage-displayed anti-idiotypic antibody single chain variable fragments to MG7 monoclonal antibody directed against XXXX ].

@entity5 :: ('MESH:D009369', 'Disease') :: ['cancer'] @entity19 :: ('10090', 'Species') :: ['mice'] @entity356 :: ('MESH:D013274', 'Disease') :: ['gastric carcinoma'] @entity526 :: ('562', 'Species') :: ['E. coli']
@entity356 :: (MESH:D013274,Disease) :: ['gastric carcinoma']
