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

Authors:
Charles E Nwankire
Charles E Nwankire
Dublin City University
Ireland
Gerard G Donohoe
Gerard G Donohoe
Dublin City University
Ireland
Xin Zhang
Xin Zhang
School of Marine Sciences
New Bedford | United States
Jonathan Siegrist
Jonathan Siegrist
University of California
United States
Martin Somers
Martin Somers
Royal College of Surgeons in Ireland
Ireland
Dirk Kurzbuch
Dirk Kurzbuch
Dublin City University
Ireland
Ruairi Monaghan
Ruairi Monaghan
Dublin City University
Ireland
Maria Kitsara
Maria Kitsara
Institute of Microelectronics
Greece

Anal Chim Acta 2013 Jun 18;781:54-62. Epub 2013 Apr 18.

Biomedical Diagnostics Institute, National Centre for Sensor Research, Dublin City University, Glasnevin, Dublin 9, Ireland.

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 30 min. The developed prototype system was used to accurately measure industrial bioprocess samples that contained 10 mg mL(-1) of hIgG.

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http://dx.doi.org/10.1016/j.aca.2013.04.016DOI Listing

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June 2013
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