Localized protein immobilization on microstructured polymeric surfaces for diagnostic applications
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- 作者：Nicole E. Steidle ; Thomas Hahn ; Christian Bader…
- 关键词：Lab ; on ; a ; chip ; Microfabrication ; Protein immobilization ; CRP
- 刊名：Microfluidics and Nanofluidics
- 出版年：2016
- 出版时间：February 2016
- 年：2016
- 卷：20
- 期：2
- 全文大小：1,147 KB
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Thomas Hahn (2)
Christian Bader (1)
Ralf Ahrens (1)
Bastian E. Rapp (1)
Andreas E. Guber (1)
Kerstin Länge (1)
1. Institute of Microstructure Technology (IMT), Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, 76344, Eggenstein-Leopoldshafen, Germany
2. Bürkert Werke GmbH & Co. KG, Christian-Bürkert-Str. 13-17, 74653, Ingelfingen, Germany - 刊物类别：Engineering
- 刊物主题：Engineering Fluid Dynamics
Medical Microbiology
Polymer Sciences
Nanotechnology
Mechanics, Fluids and Thermodynamics
Engineering Thermodynamics and Transport Phenomena - 出版者：Springer Berlin / Heidelberg
- ISSN：1613-4990
文摘
We demonstrate a localized protein immobilization method based on controlled physical adsorption on the three-phase boundary of an aqueous phase, a gas phase, and a polymeric material. By imprinting micrometer and sub-micrometer pillars onto a polymeric foil, superhydrophobic surfaces are fabricated. Those structures force the fluid locally into the Cassie–Baxter state and generate an artificial three-phase boundary at the edges of the imprinted pillars. First, fluorescence-labeled bovine serum albumin (BSA) and streptavidin dissolved in various buffer solutions are utilized to investigate protein adsorption on the structured surfaces. A stable adsorption of the respective protein on the three-phase boundary is observed. The following experiments use streptavidin adsorbed on the pillars to immobilize biotinylated antibodies for analyte detection. The pillars are passivated with an excess concentration of BSA to reduce nonspecific protein adsorption. Implemented in a lab-on-a-chip device, the proposed immobilization method is utilized in a sandwich assay to detect the inflammation marker C-reactive protein in human serum, showing the potential of this immobilization method for diagnostic applications. The method overcomes laborious procedures to immobilize proteins on thermoplastic materials, which enables the fast transfer of point-of-care applications from research to commercial scale. Keywords Lab-on-a-chip Microfabrication Protein immobilization CRP