A chemiluminescent metalloimmunoassay based on silver deposition on colloidal gold labels.
Anal Biochem
; 359(2): 247-52, 2006 Dec 15.
Article
en En
| MEDLINE
| ID: mdl-17083912
ABSTRACT
A sensitive chemiluminescent (CL) immunoassay of human immunoglobulin (IgG) which combined the inherent high sensitivity of CL analysis with the dramatic signal amplification of silver precipitation on colloidal gold tags was developed. First, the sandwich-type complex was formed in this protocol by the primary antibody immobilized on the polystyrene wells, the analyte in the sample, and the secondary antibody labeled with colloidal gold. Second, the colloidal gold was treated by an Ag(+) reduction solution, which resulted in the catalytic precipitation of silver on the surface of colloidal gold. Third, a large number of Ag(+) were oxidatively released in HNO(3) solution from the silver metal anchored on the sandwich-type complexes and then the human IgG was indirectly determined by a sensitive combined CL reaction of Ag(+)-K(2)S(2)O(8)-Mn(2+)- H(3)PO(4)-luminol. The chemiluminescence intensity depends linearly on the logarithm of the concentration of human IgG over the range of 0.02-50ngml(-1) and detection limit (3sigma) is 0.005ngml(-1) (i.e., approximately 3x10(-14)M, 3amol in 100-mul sample). This assay has been successfully applied to the determination of human IgG in human serum samples and showed great potential for numerous applications in immunoassay.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Plata
/
Inmunoglobulina G
/
Inmunoensayo
/
Técnicas Biosensibles
/
Materiales Biocompatibles Revestidos
/
Oro
/
Mediciones Luminiscentes
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Anal Biochem
Año:
2006
Tipo del documento:
Article