Journal article
Preconcentration and detection of the phosphorylated forms of cardiac troponin I in a cascade microchip by cationic isotachophoresis
Lab on a chip, Vol.11(22), pp.3793-3801
11/21/2011
Handle:
https://hdl.handle.net/2376/105363
PMCID: PMC3233477
PMID: 21935555
Abstract
This paper describes the detection of a cardiac biomarker, cardiac troponin I (cTnI), spiked into depleted human serum using cationic isotachophoresis (ITP) in a 3.9 cm long poly(methyl methacrylate) (PMMA) microfluidic channel. The microfluidic chip incorporates a 100x cross-sectional area reduction, including a 10x depth reduction and a 10x width reduction, to increase sensitivity during ITP. The cross-sectional area reductions in combination with ITP allowedvisualization of lower concentrations of fluorescently labeled cTnI. ITP was performed in both “peak mode” and “plateau mode” and the final concentrations obtained were linear with initial cTnI concentration. We were able to detect and quantify cTnI at initial concentrations as low as 46 ng mL
−1
in the presence of human serum proteins and obtain cTnI concentrations factors as high as ~ 9000. In addition, preliminary ITP experiments including both labeled cTnI and labeled protein kinase A (PKA) phosphorylated cTnI were performed to visualize ITP migration of different phosphorylated forms ofcTnI. The different phosphorylated states of cTnI formed distinct ITP zones between the leading and terminating electrolytes. To our knowledge, this is the first attempt at using ITP in a cascade microchip to quantify cTnI in human serum and detect different phosphorylated forms.
Metrics
13 Record Views
Details
- Title
- Preconcentration and detection of the phosphorylated forms of cardiac troponin I in a cascade microchip by cationic isotachophoresis
- Creators
- Danny Bottenus - Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, P.O. Box 642710, Pullman, WA, 99164-2710, U.S.AMohammad Robiul Hossan - School of Mechanical and Materials Engineering, Washington State University, Pullman, WA 99163, U.S.AYexin Ouyang - Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, P.O. Box 642710, Pullman, WA, 99164-2710, U.S.AWen-Ji Dong - School of Mechanical and Materials Engineering, Washington State University, Pullman, WA 99163, U.S.APrashanta Dutta - School of Mechanical and Materials Engineering, Washington State University, Pullman, WA 99163, U.S.ACornelius F Ivory - Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, P.O. Box 642710, Pullman, WA, 99164-2710, U.S.A
- Publication Details
- Lab on a chip, Vol.11(22), pp.3793-3801
- Academic Unit
- Chemical Engineering and Bioengineering, School of; Mechanical and Materials Engineering, School of
- Grant note
- T32 GM008336-22 || GM / National Institute of General Medical Sciences : NIGMS T32 GM008336-21 || GM / National Institute of General Medical Sciences : NIGMS
- Identifiers
- 99900546568801842
- Language
- English
- Resource Type
- Journal article