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- E-ISSN 2288-8985
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Bioconjugation by dual heterobifunctional coupling method:Use of the conjugates for the detection of dopamine
Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
2010, v.23 no.6, pp.537-543
https://doi.org/10.5806/AST.2010.23.6.537
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(2010). Bioconjugation by dual heterobifunctional coupling method:Use of the conjugates for the detection of dopamine. Analytical Science and Technology, 23(6), 537-543, https://doi.org/10.5806/AST.2010.23.6.537
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Abstract
Dopamine (DA) is an important neurotransmitter molecule of catecholamines. Its deficiency could lead to brain disorder such as Parkinson's disease and schizophrenia. Therefore, it is necessary to establish a suitable analytical technique with sensitivity and simplicity. A competitive enzyme-linked immunosorbent assay for DA has been optimized and characterized. Assay sensitivity is controlled by two factors in competitive immunoassay. One is a nature and concentration of competitor, and the other is those of binder, antibody. Thus,optimization was performed: BSA-DA conjugate and antibody-avidin conjugate were prepared by dual heterobifunctional coupling method using SATA and SMCC. Assay condition was optimized with 6.66 μgmL−1 of BSA-DA and 4.17×10−10 M of antibody-avidin conjugate. A dose-response curve was constructed, and a limit of detection and a dynamic range for DA were accomplished to 2.3×10−2 μg mL−1 and four orders of magnitude (1.0×10−7 M to 1.0×10−3 M), respectively. Calibration curve was constructed on dynamic range and least-squares regression of this data gave the following relationship: absorbance = -0.1098 log[DA]+0.0353 (R2 = 0.9956).
- keywords
- immunoassay, dopamine, heterobifunctional coupling, avidin/biotin
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