title: Polyglycerol based coatings to reduce non-specific protein adsorption in sample vials and on SPR sensors.
abstract: Coatings based on dendritic polyglycerol (dPG) were investigated for their use to control nonspecific protein adsorption in an assay targeted to analyze concentrations of a specific protein. We demonstrate that coating of the sample vial with dPG can significantly increase the recovery of an antibody after incubation. First, we determine the concentration dependent loss of an antibody due to nonspecific adsorption to glass via quartz crystal microbalance (QCM). Complementary to the QCM measurements, we applied the same antibody as analyte in an surface plasmon resonance (SPR) assay to determine the loss of analyte due to nonspecific adsorption to the sample vial. For this purpose, we used two different coatings based on dPG. For the first coating, which served as a matrix for the SPR sensor, carboxyl groups were incorporated into dPG as well as a dithiolane moiety enabling covalent immobilization to the gold sensor surface. This SPR-matrix exhibited excellent protein resistant properties and allowed the immobilization of amyloid peptides via amide bond formation. The second coating which was intended to prevent nonspecific adsorption to glass vials comprised a silyl moiety that allowed covalent grafting to glass. For demonstrating the impact of the vial coating on the accuracy of an SPR assay, we immobilized amyloid beta (Aβ) 1-40 and used an anti-Aβ 1-40 antibody as analyte. Alternate injection of analyte into the flow cell of the SPR device from uncoated and coated vials, respectively gave us the relative signal loss (1-RUuncoated/RUcoated) caused by the nonspecific adsorption. We found that the relative signal loss increases with decreasing analyte concentration. The SPR data correlate well with concentration dependent non-specific adsorption experiments of the analyte to glass surfaces performed with QCM. Our measurements show that rendering both the sample vial and the sensor surface is crucial for accurate results in protein assays.
authors: Freie Universität Berlin, Institute of Chemistry and Biochemistry, Takustr. 3, 14195 Berlin, Germany. Electronic address: tobias.becherer@fu-berlin.de.
authors: Tobias
authors: T
authors: Becherer
authors: Freie Universität Berlin, Institute of Chemistry and Biochemistry, Takustr. 3, 14195 Berlin, Germany.
authors: Christian
authors: C
authors: Grunewald
authors: Freie Universität Berlin, Institute of Chemistry and Biochemistry, Takustr. 3, 14195 Berlin, Germany.
authors: Vivienne
authors: V
authors: Engelschalt
authors: McGill University, Department of Pharmacology & Therapeutics, Faculty of Medicine, 3655 Promenade Sir-William-Osler, H3G 0B1 Montréal, QC, Canada.
authors: Gerhard
authors: G
authors: Multhaup
authors: Freie Universität Berlin, Institute of Chemistry and Biochemistry, Takustr. 3, 14195 Berlin, Germany.
authors: Thomas
authors: T
authors: Risse
authors: Freie Universität Berlin, Institute of Chemistry and Biochemistry, Takustr. 3, 14195 Berlin, Germany.
authors: Rainer
authors: R
authors: Haag
mesh_terms: D016229:Amyloid beta-Peptides / Q000032:analysis* / Q000737:chemistry* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D005898:Glass / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry*; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry* / Q000276:immunology; D011108:Polymers / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D016229:Amyloid beta-Peptides; D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D011108:Polymers; C078592:amyloid beta-protein (1-40); C043941:polyglycerol; D005990:Glycerol
keywords: Biosensor; Molecular diagnostics; Nonspecific protein adsorption; QCM; SPR