Ultrasensitive electrochemical detection of biotin using electrically addressable site-oriented antibody immobilization approach via aminophenyl boronic acid.
Resource
Biosensors & Bioelectronics,26(3),1021-1027.
Journal
Biosensors and Bioelectronics
Journal Volume
26
Journal Issue
3
Pages
1021-1027
Date Issued
2010-11
Date
2010-11
Author(s)
Abstract
A common approach towards developing immunoassays is to attach antibodies onto the surfaces of assay devices via a solid support. When directly adsorbed onto surfaces, however, antibodies generally adopt random orientations and therefore, often fail to exhibit their immunoaffinity. To preserve the antigen-binding activity of antibodies, there is an urgent need to develop specific and novel linking chemistries for attaching the antibodies to the solid surfaces in an oriented manner. In this paper, we report 2 alternative immobilization methods to enhance the orientation of antibodies onto screen-printed graphite electrodes (SPGEs). The first approach involves the deposition of gold nanoparticles (AuNPs) onto the SPGE and subsequent adsorption of monovalent half-antibody (monoAb) fragments of the anti-biotin antibody via Au-thiol bonds. For the second technique, we exploited the affinity of boronic acid towards sugar moieties by preparing a boronic acid-presenting SPGE surface to interact with the carbohydrate unit of this anti-biotin antibody. Using such approaches, we prepared an ultrasensitive electrochemical immunosensor, possessing a maximized epitope density, for the detection of biotin at concentrations as low as 0.19 pg. © 2010 Elsevier B.V.
Subjects
Aminophenyl boronic acid; Electrochemistry; Immunosensor; Liposome; Site-specific antibody immobilization
Other Subjects
Aminophenyl boronic acids; Antibody immobilization; Antigen-binding activities; Boronic acid; ELectrochemical detection; Electrochemical immunosensors; Gold nanoparticle; Immobilization method; Immunoaffinity; Random orientations; Screen-printed; Site-specific; Solid supports; Solid surface; Sugar moiety; Ultrasensitive; Acids; Adsorption; Carbohydrates; Chemical detection; Coenzymes; Electrochemistry; Gold; Gold coatings; Gold deposits; Graphite electrodes; Immunosensors; Sugar (sucrose); Antibodies; aminophenyl boronic acid; antibody; biotin; boronic acid derivative; gold nanoparticle; liposome; monovalent half antibody fragment; unclassified drug; adsorption; analytic method; antibody affinity; antigen binding; article; binding site; bioluminescence; controlled study; electrochemistry; environmental temperature; glycosylation; immunoassay; immunosensor; molecular interaction; scanning electron microscopy; Animals; Antibodies, Immobilized; Biosensing Techniques; Biotin; Boronic Acids; Electrochemical Techniques; Gold; Graphite; Humans; Immunoassay; Liposomes; Metal Nanoparticles; Mice; Nanotechnology; Sensitivity and Specificity
Type
journal article