July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Validation of a Novel Cytokine Multiplex Assay for Vitreous Analysis in Diabetic Retinopathy
Author Affiliations & Notes
  • Markus Zumbansen
    AYOXXA Biosystems GmbH, Cologne, Germany
  • Sui-Ping Ong
    AYOXXA Living Health Technologies Pte. Ltd, Singapore, Singapore
  • Kelvin Yi Chong Teo
    Singapore National Eye Centre, Singapore, Singapore
    Singapore Eye Research Institute, Singapore, Singapore
  • Marcus Ang
    Singapore National Eye Centre, Singapore, Singapore
    Singapore Eye Research Institute, Singapore, Singapore
  • Tanja Endermann
    AYOXXA Biosystems GmbH, Cologne, Germany
  • Verena Arndt
    AYOXXA Biosystems GmbH, Cologne, Germany
  • Shu Yen Lee
    Singapore National Eye Centre, Singapore, Singapore
    Singapore Eye Research Institute, Singapore, Singapore
  • Ian Yeo
    Singapore National Eye Centre, Singapore, Singapore
    Singapore Eye Research Institute, Singapore, Singapore
  • Edmund Yick Mun Wong
    Singapore National Eye Centre, Singapore, Singapore
    Singapore Eye Research Institute, Singapore, Singapore
  • Gavin Tan
    Singapore National Eye Centre, Singapore, Singapore
    Singapore Eye Research Institute, Singapore, Singapore
  • Anna Tan
    Singapore National Eye Centre, Singapore, Singapore
    Singapore Eye Research Institute, Singapore, Singapore
  • Yasuo Yanagi
    Singapore National Eye Centre, Singapore, Singapore
    Singapore Eye Research Institute, Singapore, Singapore
  • Tien Y Wong
    Singapore National Eye Centre, Singapore, Singapore
    Singapore Eye Research Institute, Singapore, Singapore
  • Nobuyo Yawata
    Singapore National Eye Centre, Singapore, Singapore
    Fukuoka Dental College, Fukuoka , Japan
  • Footnotes
    Commercial Relationships   Markus Zumbansen, AYOXXA Biosystems GmbH (E); Sui-Ping Ong, 1AYOXXALiving Health Technologies Pte. Ltd (E); Kelvin Yi Chong Teo, None; Marcus Ang, None; Tanja Endermann, AYOXXA Biosystems GmbH (E); Verena Arndt, AYOXXA Biosystems GmbH (E); Shu Yen Lee, None; Ian Yeo, None; Edmund Yick Mun Wong, None; Gavin Tan, None; Anna Tan, None; Yasuo Yanagi, None; Tien Wong, None; Nobuyo Yawata, None
  • Footnotes
    Support  NMRC/MOHIAF Cat1/0019/2014
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 5362. doi:
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      Markus Zumbansen, Sui-Ping Ong, Kelvin Yi Chong Teo, Marcus Ang, Tanja Endermann, Verena Arndt, Shu Yen Lee, Ian Yeo, Edmund Yick Mun Wong, Gavin Tan, Anna Tan, Yasuo Yanagi, Tien Y Wong, Nobuyo Yawata; Validation of a Novel Cytokine Multiplex Assay for Vitreous Analysis in Diabetic Retinopathy. Invest. Ophthalmol. Vis. Sci. 2018;59(9):5362.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Vascular endothelial growth factor (VEGF) and also cytokines and chemokines have been identified as key players in disease development and progression of diabetic retinopathy (DR) and other retinal angiogenic diseases. As such, these factors may not only represent therapeutic targets but also biomarkers for patient stratification or treatment monitoring.
However, the analysis of biomarkers from ocular fluids in the clinical setting is limited by low sample quantities. This study describes the analytical validation of a novel bead based microarray for the simultaneous quantification of IL-1β, IL-2, IL-6, IL-8, TNFα, VEGF-A from only minute amounts of ocular fluids. Furthermore, we show the analysis of vitreous humour samples from a small cohort of Diabetes mellitus (DM) patients with and without DR.

Methods : The AYOXXA LunarisTM System enables reliable multiplex analysis from minute amounts of biological samples. Capture antibody coated microspheres are immobilized in micro-sized cavities of a biochip. The assay follows the sandwich ELISA principle. By using fluorescence-labeled detection reagents signal intensity of every single reaction per microsphere is recorded using the AYOXXA reader and concentrations calculated with the Lunaris™ Analysis Software. For the analysis, 5 µl of prediluted vitreous humour samples were subjected to the standard workflow with the Lunaris™ Human 6-plex Ophthalmology Kit.

Results : The novel multiplex assay demonstrated high sensitivity, precision and accuracy within and between assay runs. In vitreous humour samples elevated levels of VEGF-A, IL-6 and IL-8 were observed in DR patients when compared to non-diabetic controls.

Conclusions : This study demonstrates the analytical reliability of the Lunaris™ platform for the evaluation of biomarkers of DR such as VEGF in small amounts of ocular fluids. This technology will enable biomarker analysis close to the origin of the disease and can become a feasible patient monitoring or stratification tool to evaluate treatment and disease progression.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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