Abstract
The limit of detection of advanced immunoassays, biochips and micro/nano biodetection devices is impacted by the non-specific adsorption of target molecules at the sample surface. In this paper, we present a simple and versatile low cost method for generating active surfaces composed of antibodies arrays surrounded by an efficient anti-fouling layer, capable to decrease drastically the fluorescence background signal obtained after interaction with a solution to be analyzed. The technological process involves the direct micro-contact printing of the antibodies probe molecules on a pre-coated PLL-g-dextran thin layer obtained by contact printing using a flat PDMS stamp. Compared to other blocking strategies (ethanolamine blocking treatment, PLL-g-PEG incubation, PLL-g-dextran incubation, printing on a plasma-deposited PEO layer), our surface chemistry method is more efficient for reducing non-specific interactions responsible for a degraded signal/noise ratio.
Highlights
Protein microarrays are used to detect and semi-quantify proteins in solution and have been used for a variety of applications (Coleman et al 2007; Gao et al 2005; Popescu et al 2007; Zhu et al 2000)
We present a simple and versatile low cost method for generating active surfaces composed of antibodies arrays surrounded by an efficient anti-fouling layer, capable to decrease drastically the fluorescence background signal obtained after interaction with a solution to be analyzed
We have investigated if direct printing of probe proteins by μCP on a PLL-g-dextran antifouling coating is possible
Summary
Protein microarrays are used to detect and semi-quantify proteins in solution and have been used for a variety of applications (Coleman et al 2007; Gao et al 2005; Popescu et al 2007; Zhu et al 2000). Antibody microarrays target essentially clinical diagnostics and proteomics fields, but their contribution is limited by: the availability of high-affinity and high-specificity antibodies for the capture of protein biomarkers, the possible protein denaturation and spatial configuration when grafted at the chip surface and the non-specific adsorption of target proteins on the surface (Hucknall et al 2009a) This last point directly impacts the biosensing performances, especially the limit of detection (LOD) (Hucknall et al 2009b). The minimal detected concentration of an immunoassay depends on the sensitivity of the signal high-concentration of a blocking molecule that does not generate any detection signal This passivating agent is intended to bind to any available surface interaction site, thereby restricting further nonspecific adsorption of the protein of interest. The dextran groups provide the required antifouling properties (Holland et al 1998; Martwiset et al 2006), which depend on its weight (Martwiset et al 2006), structure (Osterberg et al 1995) and grafting density
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