Reduced toxicity of expression, in Escherichia coli, of antipollutant antibody fragments and their use as sensitive diagnostic molecules

Citation
G. Strachan et al., Reduced toxicity of expression, in Escherichia coli, of antipollutant antibody fragments and their use as sensitive diagnostic molecules, J APPL MICR, 87(3), 1999, pp. 410-417
Citations number
33
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology,Microbiology
Journal title
JOURNAL OF APPLIED MICROBIOLOGY
ISSN journal
1364-5072 → ACNP
Volume
87
Issue
3
Year of publication
1999
Pages
410 - 417
Database
ISI
SICI code
1364-5072(199909)87:3<410:RTOEIE>2.0.ZU;2-Y
Abstract
Single-chain antibody; fragments (scAb), specific fur the chlorophenoxy aci d herbicide mecoprop, have been expressed and purified from the bacterium E scherichia coli. Co-expression with the colE1-compatible, arabinose-inducib le, skp expression vector pHELP1 prevented bacterial lysis and significantl y increased both total and functional expression yield. The periplasmic pro tein, SKP, may have a role as a generic detoxification protein. Surface pla smon resonance (BIAcore 2000) analysis confirmed that the purified scAb ret ained similar binding kinetics to the monoclonal antibody (Mab) front which it was cloned. In competition ELISA, the bacterial scAb showed the same sp ecificity for mecoprop and a related herbicide, MCPA, as the Mab but an inc rease in sensitivity for free antigen in all ELISA. formats. Bacterially expressed antibody fragments provide a simple, sensitive and co st-effective alternative to the traditional production of diagnostic Mabs v ia tissue culture.