Novel assay format permitting the prolonged use of regeneration-based sensor chip technology.
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).
Dillon
Paul P
PP
School of Biotechnology, National Centre for Sensor Research (NCSR), Dublin City University, Dublin 9, Ireland.
Killard
Anthony J
AJ
Daly
Stephen J
SJ
Leonard
Paul
P
O'Kennedy
Richard
R
D000906:Antibodies / Q000276:immunology; D006556:Heroin Dependence / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D046208:Microchip Analytical Procedures; D009021:Morphine Dependence / Q000175:diagnosis* / Q000276:immunology; D009022:Morphine Derivatives / Q000032:analysis* / Q000276:immunology; D015813:Substance Abuse Detection / Q000379:methods*; D020349:Surface Plasmon Resonance
D000906:Antibodies; D009022:Morphine Derivatives; C018108:morphine-3-glucuronide