The isolation of super-sensitive anti-hapten antibodies from combinatorialantibody libraries derived from sheep

Citation
K. Charlton et al., The isolation of super-sensitive anti-hapten antibodies from combinatorialantibody libraries derived from sheep, BIOSENS BIO, 16(9-12), 2001, pp. 639-646
Citations number
25
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biotecnology & Applied Microbiology
Journal title
BIOSENSORS & BIOELECTRONICS
ISSN journal
0956-5663 → ACNP
Volume
16
Issue
9-12
Year of publication
2001
Pages
639 - 646
Database
ISI
SICI code
0956-5663(200112)16:9-12<639:TIOSAA>2.0.ZU;2-D
Abstract
The complexity and expense of producing anti-hapten monoclonals via the tra ditional hybridoma route and the preferential selection of antibodies that recognise the conjugated form of the hapten, over antibodies that specifica lly recognise free hapten, are two of the more important problems that have limited the development and application of anti-hapten antibodies. The adv ent of phage display technology allows the rapid isolation of monoclonal an tibody fragments from libraries of different antibodies ( > 10(8)) displaye d on the surface of filamentous bacteriophages. Much of the power of this n ew approach lies in the flexibility with which these libraries can be scree ned for suitable binders. Using an optimised selection procedure, we have i solated from a sheep antibody phage display library, super-sensitive anti-h apten antibodies specific for the herbicide and environmental pollutant, at razine. In particular, two phage clones have been isolated that can be expr essed cheaply and in quantity in Escherichia coli demonstrate excellent sta bility in nonphysiological conditions and are exciting prospects for immuno assay applications including ELISA, dip-stick formats, on-line monitoring a nd biosensor technologies. In ELISA formats they show low levels of cross r eactivity with related molecules and a limit of detection of a 1-2 parts pe r trillion (p.p.t.), well within the 100 p.p.t. required by EC legislation. (C) 2001 Elsevier Science B.V. All rights reserved.