Journal
ANALYTICAL BIOCHEMISTRY
Volume 538, Issue -, Pages 13-19Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2017.09.008
Keywords
Amperometric immunosensor; Electrochemistry; Horse specific antigen; Horse IgG3; Human IgE specific assay; Anaphylactic reaction; Antivenom
Funding
- Brazilian Council for Scientific Research (CNPq) [467.488.2014-2]
- Carlos Chagas Filho Foundation of Research Support of the State of Rio de Janeiro (FAPERJ) [170.009-2008]
- Higher Education Coordination of Improvement (CAPES) [291910]
- FIOCRUZ
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Antivenom allergy disease mediated by patient IgE is an important public health care concern. To improve detection of hypersensitive individuals prior to passive antibody therapy, an amperometric immunosensor was developed to detect reactive human IgE. Whole horse IgG3 (holgG3) was immobilized onto the surface of carbon or gold screen-printed electrodes through a cross-linking solution of glutaraldehyde on a chitosan film. Sera from persons with a known allergic response to hoIgG3 or non allergic individuals was applied to the sensor. Bound human IgE (humIgE) was detected by an anti-humIgE antibody through a quantitative amperometric determination by tracking via the electrochemical reduction of the quinone generated from the hydroquinone with the application of a potential of 25 mV. The optimal immunosensor configuration detected reactive humIgE at a dilution of 1:1800 of the human sera that represent a detection limit of 0.5 pg/mL. Stability testing demonstrated that through 20 cycles of a scan, the specificity and performance remained robust. The new immunosensor successfully detected humIgE antibodies reactive against holgG3, which could allow the diagnosis of potential allergenic patients needing therapeutic antivenom preparations from a horse. (C) 2017 Published by Elsevier Inc.
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