Anti‐digoxin scFv fragments expressed in bacteria and in insect cells have different antigen binding properties

FEBS Letters

Montavon, Céline¹  Devauchelle, Gérard³  Pugnière, Martine¹  Chardès, Thierry³  Biard-Piechaczyk, Martine¹  Lemeulle, Corinne¹  Mani, Jean-Claude¹  Cerutti, Martine³  Chaabihi, Hassan²  Pau, Bernard¹ 

[1] CNRS UMR 9921, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier Cedex 2, France;Quantum Bioprobe, 26 bis rue Kléber, 93100 Montreuil, France;INRA-CNRS URA 2209, Laboratoire de Pathologie Comparée, 30380 Saint-Christol-Lez-Alès, France

关键词: Single-chain variable fragment;    Digoxin;    Affinity;    Specificity;    Baculovirus;    Escherichia coli;    Protein expression;    SPR analysis;    BCA;    bicinchoninic acid;    CNBr;    cyanogen bromide;    EDTA;    ethylenediaminetetraacetic acid;    IMAC;    ion metal chelate affinity chromatography;    IPTG;    isopropyl-thiogalactoside;    PBS;    phosphate-buffered saline;    PCR;    polymerase chain reaction;    SN;    supernatant;   

DOI  :  10.1016/S0014-5793(98)00029-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown