Abstract
BackgroundThe ability to accurately measure multiple proteins simultaneously in a single assay has the potential to markedly improve the efficiency of a myriad of clinical assays. Here, we tested the performance of a new, multiplex protein array platform to quantitate three bladder cancer-associated proteins in urine samples. The following analytes, interleukin 8 (IL8), matrix metallopeptidase 9 (MMP9), and vascular endothelial growth factor A (VEGFA) were monitored using Q-plex, a customized multiplex ELISA system from Quansys Biosciences, and individual target commercial ELISA kits. The performance of the two approaches was compared by evaluating the diagnostic accuracy of the biomarker assays in samples from a cohort of 73 subjects of known bladder cancer status.ResultsThe combination biomarker panel analyses revealed an AUROC value of 0.9476 for the Q-plex assay, and 0.9119 for the combination of the single-target ELISA assays. The Q-plex assay achieved an overall diagnostic sensitivity of 0.93 and specificity of 0.81, and the individual target ELISA assays achieved an overall sensitivity of 0.77 and specificity of 0.91.ConclusionBased on these encouraging preliminary data, we believe that the Q-Plex technology is a viable platform that can be exploited as an efficient, highly accurate tool to quantitate multiplex panels of diagnostic proteins in biologic specimens.
Highlights
The ability to accurately measure multiple proteins simultaneously in a single assay has the potential to markedly improve the efficiency of a myriad of clinical assays
Technologies that can efficiently, simultaneously monitor molecular signatures would be pivotal in moving promising biomarker panels towards clinical utility
Quansys Biosciences have developed the Q-Plex, a multiplex Enzyme-Linked Immunosorbent Assays (ELISA) system that prints capture antibodies as an array in a single well
Summary
The ability to accurately measure multiple proteins simultaneously in a single assay has the potential to markedly improve the efficiency of a myriad of clinical assays. The following analytes, interleukin 8 (IL8), matrix metallopeptidase 9 (MMP9), and vascular endothelial growth factor A (VEGFA) were monitored using Q-plex, a customized multiplex ELISA system from Quansys Biosciences, and individual target commercial ELISA kits. The objective of the current feasibility study was to test the ability of a customized multiplex assay system to accurately and simultaneously monitor three urinary protein biomarkers from one of our validated bladder cancer signatures [8]. The levels of the three proteins, interleukin 8 (IL8), matrix metallopeptidase 9 (MMP9), and vascular endothelial growth factor A (VEGFA), were quantitated in real-world urine samples and the diagnostic performance of the multiplex system was compared with analyses performed using three individual target commercial ELISA kits. Chemiluminescent ELISAs have been shown to be more sensitive than colorimetric detection systems [11,12]
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