Journal
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
Volume 43, Issue -, Pages 39-48Publisher
WILEY
DOI: 10.1042/BA20050073
Keywords
antibody; biodistribution; carcinoembryonic antigen (CEA); diabody; Escherichia coli; single-chain Fv (scFv)
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In the present paper we report the development of a bivalent scFv (single-chain Fv) antibody fragment, starting from a mouse mAb (monoclonal antibody) specific for CEA (carcinoembryonic antigen) that has received approval for in vivo radioimmunodiagnosis in humans. The diabody is well expressed in Escherichia coli, is easily purified by a combination of immobilized metal ion affinity chromatography and gel filtration and exhibits high affinity and specificity for CEA, comparable with those of the original mAb. Biodistribution experiments in athymic nude mice transplanted with human CEA+ cancer cells showed that the I-125-labelled diabody preferentially localizes in the tumour tissue and that retention is still high 48 h after injection. The diabody can be advantageous for some in vivo tumour targeting applications, due to the faster clearance derived from its smaller molecular mass.
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