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Novel assay format permitting the prolonged usenext term of regeneration-based sensor chip technology

Dillon, Paul P. and Killard, Anthony J. and Daly, Stephen J. and Leonard, Paul and O'Kennedy, Richard (2004) Novel assay format permitting the prolonged usenext term of regeneration-based sensor chip technology. Journal of Immunological Methods, 296 (1-3). pp. 77-82. ISSN 0022-1759

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Abstract

A polyclonal antibody raised against morphine-3-glucuronide (M3G, the main metabolite of heroin and morphine) was used in the development of a novel assay format using a surface plasmon resonance (SPR)-based biosensor. Previously developed assays have generated calibration curves based on differences in the quantity of response units binding to the surface of a chip coated with the analyte. The novel assay described here was based on the development of a standard curve using the slope of a series of consecutive binding interactions. Using this format, regeneration between each assay cycle was no longer required. This increased the useable life span of the chip surface and, as a result, decreased the cost associated with the assay. Thus, at least 15 binding interactions could be carried out before the saturation of antibody on the surface of the chip caused the response to deviate significantly from linearity. After 15 nonregenerated binding interactions, the slope still remained within 1.5% of the slope after a single binding event. Analysis time, and the sample volumes required were also markedly decreased while sensitivity was enhanced. The inhibition assay developed had a detection range of 270 to 17,500 pg ml 1 .

Item Type:Article (Published)
Refereed:Yes
Uncontrolled Keywords:morphine-3-glucuronide; surface plasmon resonance; biosensor; inhibition assay;
Subjects:Biological Sciences > Biotechnology
Physical Sciences > Chemistry
Biological Sciences > Biosensors
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Chemical Sciences
DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology
Research Initiatives and Centres > National Centre for Sensor Research (NCSR)
Publisher:Elsevier
Official URL:http://dx.doi.org/10.1016/j.jim.2004.10.013
ID Code:50
Deposited On:08 Nov 2006 by DORAS Administrator. Last Modified 30 Jan 2009 11:50

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