Anti-Clostridium tetani Antibody Determination in Serum Samples by Amperometric Immunosensing

An electrochemical (EC) immunosensing assay for anti‐Clostridium tetani antibody determination in serum has been developed. The antigen tetanus toxoid was immobilized on superparamagnetic nanobeads. The immunoreaction occurred in Eppendorf minitubes. The anti‐tetani antibody was incubated in the pre...

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Bibliographic Details
Published in:Electroanalysis (New York, N.Y.) N.Y.), 2010-01, Vol.22 (1), p.41-48
Main Authors: Patris, S., De Vriese, C., Prohoroff, F., Calvo, E. Burgoa, Martínez, J. Arcos, Kauffmann, J.-M.
Format: Article
Language:English
Subjects:
Antibodies
Clostridium tetani
Immunoassays
Immunosensing
Screen printed electrode
Tetani antibody
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Summary:An electrochemical (EC) immunosensing assay for anti‐Clostridium tetani antibody determination in serum has been developed. The antigen tetanus toxoid was immobilized on superparamagnetic nanobeads. The immunoreaction occurred in Eppendorf minitubes. The anti‐tetani antibody was incubated in the presence of the toxoid functionalized nanobeads, then reacted with horseradish peroxidase‐labeled anti‐IgG. The resulting immunobeads were retained onto the carbon paste working electrode with a magnet. Hydroquinone served as redox label. The level of anti‐Clostridium tetani antibody in guinea pig serum samples was determined by amperometry using a carbon based screen‐printed electrode (cSPE) housed onto a magnetic support. The EC response was proportional to the logarithm of the antibody concentration comprised between 0.0046 IU/mL and 0.175 IU/mL with a limit of detection of 0.0046 IU/mL. In order to minimize the matrix effect, the standard addition method was applied. The assay was validated by comparing the EC immunosensing data with those obtained by applying the ELISA method described in the European Pharmacopoeia.
ISSN:1040-0397
1521-4109
DOI:10.1002/elan.200900396