Microfabricated On-Chip-Type Electrochemical Flow Immunoassay System for the Detection of Histamine Released in Whole Blood Samples

Abstract

This paper describes an on-chip-type electrochemical flow immunoassay system with a multichanneled matrix column. The multichanneled matrix column was functionally coated with cation-exchange resin and used for separation of proteins. Antihistamine immunoglobulin G (IgG) antibody conjugated with ferrocenemonocarboxylic acid (Fc) was also prepared and used as a novel analytical reagent. Antibody-antigen complexes were separated from free Fc-conjugated IgG antibody (Fc-IgG) on the basis of differences in isoelectric point (pI) using the multichanneled matrix column coated with cation-exchange resin. The assay yields a good relationship between current and histamine concentration in the range of 200-2000 ng/mL. This simple technique enables the assay of histamine released in whole blood within 2 min. Furthermore, a good correlation was found between the response of the electrochemical immunoassay described in this paper and the conventional RIA (radioimmunoassay). This on-chip-type electrochemical flow immunoassay requires only minute quantities of whole blood samples and generates highly reproducible results.

Recent developments in electrochemical detection for microchip capillary electrophoresis

“…Working electrodes used for the microchip format have been fabricated from several materials, including carbon fiber [44,[46][47][48], carbon paste [49,50], carbon ink [15,39,45,[51][52][53], glassy carbon [54], photoresist carbon films [55,56], gold [13,14,27,31,32,43,52,[57][58][59][60][61][62][63][64][65][66], platinum [32,[67][68][69][70][71], copper [32,46,59,72], palladium [25], and diamond [73,74].…”

“…Several analytes have been detected with carbon-based electrodes, including catechols [44][45][46], neurotransmitters [44-48, 53, 55], ascorbic acid [53,55], amino acids [15,51,75], carbohydrates [14], peptides [15], phenols [75], nucleotides [56], histamine [54], and nitroaromatic compounds [52]. The majority of carbon-based electrodes that have been used on-chip are composed of carbon fibers, paste, or ink.…”

PVC‐Based on Thiopyrilium Derivatives Membrane Electrodes for Determination of Histamine

“…The development of analytical methodology for the determination of histamine is an important subject. There have been many methods for measuring histamine, including high performance liquid chromatography (HPLC), 5,6 double or single-isotope radioenzymatic assays, 7 electrochemical flow immunoassay, 8 gas chromatography-mass spectrometry, 9 enzyme-based amperometric biosensors using histamine oxidase, 10 methylamine dehydrogenase, 11,12 quinohemoprotein amine dehydrogenase 13 capillary electrophoresis combined with UV detector 14 and carbon fiber-based electrochemical techniques. 15 Most of these techniques, however, suffer from disadvantages such as derivatization, expensive instrumentation, low specificity, linearity, sensitivity and interference from molecular oxygen.…”

Challenges in Developing a Biochip for Intact Histamine Using Commercial Antibodies

“…Finally, histamine content in fermented vegetable juices was determined. Electrochemical flow immunoassay for histamine in blood samples was established using monoclonal anti-histamine antibody purchased from Biogenesis [33]. In the competitive immunoreaction histamine-IgG complex was separated from the histamine-BSA (coupled using 1,4-benzoquinone)-IgG complex, and the current change was measured using a multichanneled matrix column.…”