Development of a novel multiplexed microarray lab-on-a-chip device to quantify ocular surface disease biomarkers in tears

Purpose: Ocular surface diseases (OSD) are the major cause of ophthalmological consultation, produce symptoms as a consequence of multiple physiological processes. Evaluation of biomarker panels involved in these biological processes might be a good strategy for diagnosis and monitoring ocular diseases. In the present study, the development of a multiplexed microarray-based device has been performed Methods: Three previously validated biomarkers for OSD (S100A6, CST4 and MMP9) were selected for the integration into a lab-on-a-chip system (Fraunhofer ivD-platform) with capacity for 500 protein spots. The integration process consisted on several steps: i) identification of specific capture and detection antibodies for biomarkers; ii) selection of an appropriate microarray surface for antibody immobilization; iii) evaluation of cross-reactivity between antibodies; iv) determination of detection limits; v) technical validation of the platform with tear samples Results: Critical parameters for protein quantification on Fraunhofer ivD-platform were established. Detection dynamic ranges of the selected antibody pairs fulfilled the requirements for quantification in tears into clinical relevance range. No crossreactivity was detected on the system. Technical validation with 1 µl of tear samples confirmed accuracy of previously reported data in ELISA assays Conclusions: A novel lab-on-a-chip system was successfully developed for simultaneous quantification of three OSD biomarkers. The device fulfilled technical requirements for clinical practice use. Additional biomarkers for other OSD might further be included, making this novel lab-on-a-chip system a potential multi-disease tool for management of OSD.