An electro-active system of immuno-assay (EASI assay) utilising self assembled monolayer modified electrodes.
Most immunoassays currently rely on optical methods for signal generation e.g. in ELISA and rapid assay formats. It has become apparent as in the Glucose sensor market that there is a need for simple direct electrical immuno-sensors. We have investigated the novel use of organic conducting monolayers used as a direct electrochemical detection support for an immuno-reaction. It was found that antibodies raised to a carbazole dimer monolayer could increase the charge movement across that monolayer surface. Antibody fragments were taken from a specific anti-carbazole antibody fragment library and combined with an antibody fragment directed to the hormone estrone 3 glucuronide (E3G), the target antigen to form a bispecific antibody fragment. The device utilised these specific antibody fragments and incorporated them on the top plate of a capillary fill format as the immuno-assay components. The immuno-reaction utilised a competition assay. Free E3G analyte in the sample displaced the bispecific antibody fragment from the immuno-surface leaving it free to bind the carbazole monolayer surface. There the binding was detected using amperometric or coulometric methods. By combining all there element it was possible to develop a sensitive immuno-assay that could detect E3G in a reproducible calibrated fashion down to 10 ng/ml.
Porter
R
R
Unilever Research Colworth, Colworth House, Sharnbrook, Bedford MK44 1LQ, UK. robert.porter@unilever.com
van der Logt
P
P
Howell
S
S
Kyröläinen-Reay
M
M
Badley
A
A
D000595:Amino Acid Sequence; D000818:Animals; D018033:Antibodies, Bispecific / Q000235:genetics; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods* / Q000706:statistics & numerical data; D002161:Camelids, New World; D002227:Carbazoles / Q000276:immunology; D004563:Electrochemistry; D004566:Electrodes; D004582:Electronics, Medical; D004797:Enzyme-Linked Immunosorbent Assay; D004970:Estrone / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D007118:Immunoassay / Q000379:methods* / Q000706:statistics & numerical data; D007128:Immunoglobulin Fragments / Q000235:genetics; D008969:Molecular Sequence Data; D012680:Sensitivity and Specificity
D018033:Antibodies, Bispecific; D002227:Carbazoles; D007128:Immunoglobulin Fragments; C015856:estrone-3-glucuronide; D004970:Estrone