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At-line bioprocess monitoring by immunoassay with rotationally controlled serial siphoning and integrated supercritical angle fluorescence optics

Authors
Journal
Analytica Chimica Acta
0003-2670
Publisher
Elsevier
Publication Date
Volume
781
Identifiers
DOI: 10.1016/j.aca.2013.04.016
Keywords
  • Lab-On-A-Disc
  • Microfluidics
  • Bioprocess Monitoring
  • Supercritical Angle Fluorescence
  • Integrated
Disciplines
  • Biology
  • Chemistry

Abstract

Abstract In this paper we report a centrifugal microfluidic “lab-on-a-disc” system for at-line monitoring of human immunoglobulin G (hIgG) in a typical bioprocess environment. The novelty of this device is the combination of a heterogeneous sandwich immunoassay on a serial siphon-enabled microfluidic disc with automated sequential reagent delivery and surface-confined supercritical angle fluorescence (SAF)-based detection. The device, which is compact, easy-to-use and inexpensive, enables rapid detection of hIgG from a bioprocess sample. This was achieved with, an injection moulded SAF lens that was functionalized with aminopropyltriethoxysilane (APTES) using plasma enhanced chemical vapour deposition (PECVD) for the immobilization of protein A, and a hybrid integration with a microfluidic disc substrate. Advanced flow control, including the time-sequenced release of on-board liquid reagents, was implemented by serial siphoning with ancillary capillary stops. The concentration of surfactant in each assay reagent was optimized to ensure proper functioning of the siphon-based flow control. The entire automated microfluidic assay process is completed in less than 30min. The developed prototype system was used to accurately measure industrial bioprocess samples that contained 10mgmL−1 of hIgG.

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