Detection of inflammatory cytokines using a fiber optic microsphere immunoassay array

Timothy M. Blicharz; David R. Walt

doi:10.1117/12.686158

23 October 2006 Detection of inflammatory cytokines using a fiber optic microsphere immunoassay array

Timothy M. Blicharz, David R. Walt

Proceedings Volume 6380, Smart Medical and Biomedical Sensor Technology IV; 638010 (2006) https://doi.org/10.1117/12.686158
Event: Optics East 2006, 2006, Boston, Massachusetts, United States

Abstract

A multiplexed fiber optic microsphere-based immunoassay array capable of simultaneously measuring five inflammatory cytokines has been developed. Five groups of amine-functionalized 3.1 micron microspheres were internally encoded with five distinct concentrations of a europium dye and converted to cytokine probes by covalently coupling monoclonal capture antibodies specific for human VEGF, IFN-gamma, RANTES, IP-10, and Eotaxin-3 to the microspheres via glutaraldehyde chemistry. The microspheres were pooled and loaded into a 1 mm diameter fiber optic bundle containing ~50,000 individual etched microwells, producing the multiplexed cytokine immunoassay array. Multiple arrays can be created from a single microsphere pool for high throughput sample analysis. Sandwich fluoroimmunoassays were performed by incubating the probe array in a sample, followed by incubation in a mixture of biotin-labeled detection antibodies that are complementary to the five cytokines. Finally, universal detection of each protein was performed using a fluorescence imaging system after briefly immersing the array in a solution of fluorophore-labeled streptavidin. The multiplexed cytokine array has been shown to respond selectively to VEGF, IFNgamma, RANTES, IP-10, and Eotaxin-3, permitting multiplexed quantitative analysis. Ultimately, the multiplexed cytokine array will be utilized to evaluate the potential of using saliva as a noninvasive diagnostic fluid for pulmonary inflammatory diseases such as asthma.

Citation Download Citation

Timothy M. Blicharz and David R. Walt "Detection of inflammatory cytokines using a fiber optic microsphere immunoassay array", Proc. SPIE 6380, Smart Medical and Biomedical Sensor Technology IV, 638010 (23 October 2006); https://doi.org/10.1117/12.686158

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