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  4. Application of surface biopassivated disposable poly(dimenthylsiloxane)/glass chips to a heterogeneous competitive human serum immunoglobulin G immunoassy with incorporated internal standard
 
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research article

Application of surface biopassivated disposable poly(dimenthylsiloxane)/glass chips to a heterogeneous competitive human serum immunoglobulin G immunoassy with incorporated internal standard

Linder, V.
•
de Rooij, N. F.  
•
Verpoorte, E.
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2002
Electrophoresis

A microfluidic platform for a heterogeneous competitive immunoassay of human immunoglobulin G (IgG) employing Cy5-human IgG as tracer and Cy3-mouse IgG as internal standard was developed. The device consisted of microchannels made of poly(dimethylsiloxane) and glass which were patterned with antibodies against human IgG and mouse IgG. Electrokinetic sample transport was employed in order to exploit the small difference between the net mobilities of analyte and tracer, thereby achieving favorable conditions for the performance of the competitive immunoreaction. The overall quality of the disposable chip and performance of the immunoassay were controlled by monitoring the fluorescence of bound tracer and bound internal standard. Analyses with an insufficient internal standard response were discarded, and immunoassay data evaluation was based on the ratio of tracer and internal standard fluorescence. Using synthetic samples in the range from 0 to 80 μg/mL IgG and alkaline running conditions, a concentration-dependent response with reproducible Cy5/Cy3 signal ratios (average relative standard deviation of 6.8%) was obtained. Chips stored with solution in the channels at 4°C over a two-month period were found to perform like freshly prepared chips, whereas chips stored dry at -20°C and rehydrated prior to use could not be employed. The analysis of patient sera showed that the immunoassay platform behaved differently in the presence of serum-based samples. Using the same conditions as for the synthetic samples, no concentration dependence was noted. With a large excess of tracer, however, an IgG concentration dependence was observed, permitting distinction of samples of patients with normal IgG serum levels (8-16 mg/mL) from those with elevated IgG concentrations (> 16 mg/mL).

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research article
DOI
10.1002/1522-2683(200203)23:5<740::AID-ELPS740>3.0.CO;2-7
Author(s)
Linder, V.
•
de Rooij, N. F.  
•
Verpoorte, E.
•
Sigrist, H.
•
Thormann, W.
Date Issued

2002

Published in
Electrophoresis
Volume

23

Issue

5

Start page

740

End page

749

Note

279

Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
SAMLAB  
Available on Infoscience
May 12, 2009
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/39553
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