The mode of action of Y13-259 scFv fragment intracellularly expressed in mammalian cells.
The anti-p21ras Y13-259 single-chain Fv fragment (scFv) neutralizes the activity of p21-ras when intracellularly expressed in different systems. We have studied the mode of action of this inhibition in 3T3 K-ras fibroblasts and demonstrated that (i) this antibody fragment is highly aggregating when cytoplasmically expressed and (ii) the p21-ras antigen is sequestered in these aggregates in an antibody-dependent manner. This co-segregation leads to an efficient inhibition of DNA synthesis. These results suggest that an antigen can be diverted from its normal location inside the cells in an antibody mediated way, prospecting a new mode of action for intracellular antibodies in vivo.
Cardinale
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A
Department of Neuroscience, University of Rome Tor Vergata, Roma, Italy.
Lener
M
M
Messina
S
S
Cattaneo
A
A
Biocca
S
S
D016475:3T3 Cells; D000818:Animals; D000912:Antibodies, Neoplasm / Q000276:immunology*; D019556:COS Cells; D002455:Cell Division; D004247:DNA / Q000096:biosynthesis; D005455:Fluorescent Antibody Technique; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology*; D051379:Mice; D016283:Proto-Oncogene Proteins p21(ras) / Q000037:antagonists & inhibitors* / Q000276:immunology* / Q000378:metabolism; D014162:Transfection
D000912:Antibodies, Neoplasm; D007128:Immunoglobulin Fragments; D004247:DNA; D016283:Proto-Oncogene Proteins p21(ras)