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Anti‐digoxin scFv fragments expressed in bacteria and in insect cells have different antigen binding properties
Author(s) -
Lemeulle Corinne,
Chardès Thierry,
Montavon Céline,
Chaabihi Hassan,
Mani Jean-Claude,
Pugnière Martine,
Cerutti Martine,
Devauchelle Gérard,
Pau Bernard,
Biard-Piechaczyk Martine
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00029-5
Subject(s) - escherichia coli , antibody , fragment (logic) , monoclonal antibody , microbiology and biotechnology , antigen , gene , chemistry , cloning (programming) , expression vector , single chain variable fragment , bacteria , vector (molecular biology) , biology , biochemistry , recombinant dna , genetics , computer science , programming language
A gene encoding a single‐chain antibody fragment directed against digoxin (named 1C10 scFv) was cloned in two expression systems. For this purpose, a new baculovirus transfer cassette fully compatible with the procaryotic pHEN vector was constructed. Baculovirus production led to higher yield than did Escherichia coli expression. The procaryotic fragment showed variations in the fine specificity profile but an affinity constant nearly identical to that of the 1C10 F ab , whereas the eucaryotic scFv fragment had a lower affinity with a specificity profile identical to original mAb. The half‐lives of the digoxin:scFv complexes and the global specificity are compatible with therapeutic use of this antibody fragment.