TITLE:
A Comparison of Enzyme-Linked Immunosorbent Assay versus Multiplex Methodology Using an in Vitro Model of Pulmonary Hypertension and Inflammation
AUTHORS:
Yan Zhu, Deepthi Alapati, Joanna Costa, Victoria L. Maduskuie, Paul T. Fawcett, Thomas H. Shaffer
KEYWORDS:
Enzyme-Linked Immunosorbent Assay (ELISA), Luminex, Pulmonary Artery Smooth Muscle Cells (PASMC), Inflammation, Bland-Altman Plot Analysis
JOURNAL NAME:
Journal of Biomedical Science and Engineering,
Vol.7 No.7,
June
6,
2014
ABSTRACT: Enzyme-linked immunosorbent assay (ELISA) is the most widely used method for measuring a single cytokine. Recent developments in cytokine quantification such as multiple arrays measure multiple cytokines simultaneously. Although good correlations between ELISA and multiplex methods have been observed, side by side comparisons are limited. In the present study we hypothesized that ELISA and Luminex techniques are comparable in detecting cytokines in culture medium when pulmonary artery smooth muscle cells (PASMC) are exposed to stress. Primary human PASMC were cultured in modular chambers and exposed to 21% FiO2 and peak inspiratory and positive end expiratory pressure of 24 and 8 cmH2O respectively, and 95% FiO2. At 24 hours, culture medium was collected and assayed for interleukin-6 (IL-6) and IL-8 by quantitative ELISA and by Human Cytokine 25-Plex Panel using a Luminex 200 analyzer. A comparative analysis of agreement between our ELISA and Luminex data was detailed for control and stress conditions using the Bland-Altman plot analysis. Each assay resulted in comparable increased (p