Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays.
A novel signal amplification method, catalyzed reporter deposition (CARD), and its application to immunoassays is described. The method involves utilizing an analyte-dependent reporter enzyme (ADRE) to catalyze the deposition of additional reporter on the surface in a solid-phase immunoassay. In the examples described, deposition of reporter is facilitated by using a horseradish peroxidase (HRP) ADRE to catalyze the deposition of biotin labeled phenols. The deposited biotins are then reacted with streptavidin-labeled enzyme, thereby resulting in deposition of enzyme. Using the ADRE to catalyze the deposition of additional enzyme results in an amplification of the signal of the ADRE alone and improves the detection limit of the assay. The method is highly sensitive, simple, flexible, and easy to implement.
Bobrow
M N
MN
Medical Products Department, E.I. Du Pont De Nemours & Co., North Billerica, MA 01862.
Harris
T D
TD
Shaughnessy
K J
KJ
Litt
G J
GJ
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D001426:Bacterial Proteins; D001710:Biotin; D004306:Dose-Response Relationship, Immunologic; D015488:HIV Antigens / Q000032:analysis; D006735:Horseradish Peroxidase; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis; D051379:Mice; D010636:Phenols; D012191:Retroviridae Proteins / Q000276:immunology; D018139:Simplexvirus / Q000276:immunology; D019809:Streptavidin
D000956:Antigens, Viral; D001426:Bacterial Proteins; D015488:HIV Antigens; D007074:Immunoglobulin G; D010636:Phenols; D012191:Retroviridae Proteins; D001710:Biotin; D019809:Streptavidin; D006735:Horseradish Peroxidase