Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9902290 | Journal of Immunological Methods | 2005 | 10 Pages |
Abstract
Intracellular antibodies or intrabodies have great potential in protein knockout strategies for intracellular antigens. We applied the Intracellular Antibody Capture Technology for the direct selection in yeast of a mouse scFv library (VL-VH format) constructed from animals immunised with recombinant non-structural protein NSP5 of Rotavirus. We selected five different intracellular antibodies (ICAbs), which specifically recognize Î2, an NSP5 deletion mutant used as bait. The anti-NSP5 ICAbs were well expressed both in yeast and mammalian cells as cytoplasmic or nuclear-tagged forms. By immunofluorescence and co-immunoprecipitation assays we characterised the intracellular interaction of the five anti-NSP5 ICAbs with the co-expressed antigens.
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Authors
Fulvia Vascotto, Michela Visintin, Antonino Cattaneo, Oscar R. Burrone,