Expression in Escherichia-Coli of Soluble and M13 Phage-Displayed Forms of a Single-Chain Antibody Fragment Specific for Digoxin - Assessment in a Novel Drug Immunoassay - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue Immunotechnology Année : 1995

Expression in Escherichia-Coli of Soluble and M13 Phage-Displayed Forms of a Single-Chain Antibody Fragment Specific for Digoxin - Assessment in a Novel Drug Immunoassay

I. Navarroteulon
  • Fonction : Auteur
S. Peraldiroux
  • Fonction : Auteur
T. Bernardi
  • Fonction : Auteur
M. Marin
  • Fonction : Auteur
D. Shire
  • Fonction : Auteur
B. Pau
  • Fonction : Auteur
M. Biardpiechaczyk
  • Fonction : Auteur

Résumé

A high affinity anti-digoxin single-chain Fv antibody fragment (scFv) was cloned from the mouse 2C2 hybridoma cell line and was functionally expressed both in the Escherichia coli periplasm as a soluble molecule and at the surface of the filamentous M13 bacteriophage as a fusion protein with the gene III minor coat protein. The 2C2 scFv sequence significantly differs from that of all the other anti-digoxin antibodies previously described. The 2C2 scFv shares with its parental monoclonal antibody a high specificity for digoxin, a cross-reactivity with active digoxin metabolites, but none with inactive metabolites, M13 phages displaying the 2C2 scFv at their surface have a high apparent affinity constant for digoxin (6.6 x 10(8) M(-1)) and were directly used to set up a novel type of immunoenzymatic assay for monitoring digoxin in sera of patients treated for either congestive heart failure or cardiac arrythmias. We thus report for the first time that phages displaying scFv may constitute a large source of important new reagents in the field of immunodiagnosis.

Dates et versions

hal-02193517 , version 1 (24-07-2019)

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Citer

I. Navarroteulon, S. Peraldiroux, T. Bernardi, M. Marin, Marc Piechaczyk, et al.. Expression in Escherichia-Coli of Soluble and M13 Phage-Displayed Forms of a Single-Chain Antibody Fragment Specific for Digoxin - Assessment in a Novel Drug Immunoassay. Immunotechnology, 1995, 1 (1), pp.41--52. ⟨10.1016/1380-2933(95)00004-6⟩. ⟨hal-02193517⟩
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