Optical heterogeneous bioassay for the detection of the inflammatory biomarker suPAR

S. Tombelli; C. Trono; B. Adinolfi; F. Chiavaioli; A. Giannetti; J. Eugen-Olsen; R. Bernini; Immacolata A. Grimaldi; G. Persichetti; G. Testa; F. Baldini

doi:10.1117/12.2082804

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4 March 2015 Optical heterogeneous bioassay for the detection of the inflammatory biomarker suPAR

S. Tombelli, C. Trono, B. Adinolfi, F. Chiavaioli, A. Giannetti, J. Eugen-Olsen, R. Bernini, Immacolata A. Grimaldi, G. Persichetti, G. Testa, F. Baldini

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Proceedings Volume 9313, Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XIII; 93130W (2015) https://doi.org/10.1117/12.2082804
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

Soluble urokinase plasminogen activator receptor (suPAR) is an inflammatory protein present in blood and a marker of disease presence, severity and prognosis. A heterogeneous sandwich assay is proposed for quantifying suPAR by employing a capture antibody from rat and a biotinylated detection antibody from mouse. Optical detection was achieved by a successive exposure of the biotinylated sandwich to streptavidin labelled with ATTO647N. The heterogeneous assay was implemented on a multichannel polymethylmetacrylate (PMMA) optical biochip, potentially capable of the simultaneous detection of more than one analyte. Capture antibody was immobilized on the PMMA surface of the microfluidic channel and the assay was performed with the following protocol: i) surface blocking with BSA, ii) incubation with suPAR or PBS, iii) incubation with biotinylated suPAR detection Ab and iv) incubation with streptavidin-ATTO647N. Promising preliminary results were obtained with this protocol. Moreover, an improved optical setup is proposed which avoids the mechanical scanning of the chip and consequently the in-series fluorescence excitation and read out, allowing the simultaneous measurement of the fluorescence on all the channels of the microfluidic chip.

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S. Tombelli, C. Trono, B. Adinolfi, F. Chiavaioli, A. Giannetti, J. Eugen-Olsen, R. Bernini, Immacolata A. Grimaldi, G. Persichetti, G. Testa, and F. Baldini "Optical heterogeneous bioassay for the detection of the inflammatory biomarker suPAR", Proc. SPIE 9313, Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XIII, 93130W (4 March 2015); https://doi.org/10.1117/12.2082804

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