Characterisation of antibody models of the ryanodine receptor for use in high-throughput screening

Citation
Aj. Dinsmore et al., Characterisation of antibody models of the ryanodine receptor for use in high-throughput screening, PEST SCI, 54(4), 1998, pp. 345-352
Citations number
16
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Entomology/Pest Control
Journal title
PESTICIDE SCIENCE
ISSN journal
0031-613X → ACNP
Volume
54
Issue
4
Year of publication
1998
Pages
345 - 352
Database
ISI
SICI code
0031-613X(199812)54:4<345:COAMOT>2.0.ZU;2-N
Abstract
The syntheses of seven novel synthetic analogues of the naturally occurring insecticide ryanodine are described. These, and other synthetic and natura lly occurring analogues, have been used to characterise the selectivity of a monoclonal antibody which has been produced by immunisation with 9-hydrox y-21-(4-azidobenzyloxy)-9-epiryanodine photo-conjugated to keyhole limpet h aemocyanin. The antibody binds [H-3]ryanodine with a dissociation constant of 0.37 nM. The specificity of this antibody in terms of its ability to rec ognise 11 natural and synthetic analogues of ryanodine has been determined by [H-3]ryanodine displacement and shown to be similar (for a partially ove rlapping set of analogues) to that determined earlier for a rabbit polyclon al antibody (Kahl, S. D., er al., Anal. Biochem., 218 (1994) 55-62). The se lectivity of the antibodies is shown to be related to that of the sarcoplas mic reticulum Ca2+ release channel from rabbit skeletal muscle, both for th is set of ryanodine analogues and for three structurally dissimilar, low-mo lecular-weight compounds identified by high-throughput screening. The advan tages of these antibody models of the Ca2+ release channel for screening ar e illustrated by their superior performance in a homogeneous binding assay. (C) 1998 Society of Chemical Industry.