Multilabeling of ferrocenes to a glucose oxidase-digoxin conjugate for the development of a homogeneous electroenzymatic immunoassay.
A new homogeneous electroenzymatic immunoassay was developed to determine antibody concentration using glucose oxidase and ferrocene as enzymatic and electrochemical amplifier, respectively. Digoxin (Dig)-conjugated glucose oxidase (GOx) was modified with ferrocene (Fec) to form Fec-GOx-Dig conjugate. After immunocomplex formation between the Fec-GOx-Dig conjugate and the anti-digoxin antibody, the complex underwent less electrochemical reaction due to the steric hindrance of the antibody. The ferrocene multilabeled conjugate was provided for the determination of anti-digoxin antibody. Since the strategy taken here is based on a combined effect of GOx and ferrocene, i.e., enzymatic amplification by GOx and electrochemical amplification by multilabeled ferrocenes, the antibody concentration was determined in the range from 1/50 to 1/500 dilution.
Suzawa
T
T
Department of Bioengineering, Tokyo Institute of Technology, Yokohama, Japan.
Ikariyama
Y
Y
Aizawa
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000378:metabolism; D004077:Digoxin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D004563:Electrochemistry; D005296:Ferrous Compounds / Q000737:chemistry / Q000378:metabolism*; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000737:chemistry; D000075163:Metallocenes; D051379:Mice; D009942:Organometallic Compounds / Q000737:chemistry / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D005296:Ferrous Compounds; D007074:Immunoglobulin G; D000075163:Metallocenes; D009942:Organometallic Compounds; D004077:Digoxin; D005949:Glucose Oxidase; C004998:ferrocene