Identification of unacceptable background caused by non-specific protein adsorption to the plastic surface of 96-well immunoassay plates using a standardized enzyme-linked immunosorbent assay procedure

Rebeski, Dierk E.; Winger, E.M; Shin, Yeun-Kyung; Lelenta, Mamadou; Robinson, Mark; Varecka, R; Crowther, John R.

doi:10.1016/s0022-1759(99)00051-4

Cited by 51 publications

(27 citation statements)

References 2 publications

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“…Among these, PS and GS are most commonly used solid materials because they readily adsorb to DNA and proteins (antibodies or antigens) via non-covalent interactions (Chu et al 2005), excellent optical as well as mechanical properties, and cost-effective (North et al 2010). However, the non-covalent interactions of DNA and proteins on their surfaces also have limitations like desorption during the time of washing, poor recognizing properties, protein denaturation, loss of bioactivity, and low sensitivity (Rebeski et al 1999;Butler 2000). To overcome these problems, covalent immobilization protocols have been developed and it ensured with the strong attachment and displaying a reorganization site of interest with respect to its target (Goddard and Hotchkiss 2007).…”

Section: Introductionmentioning

confidence: 99%

Gold nanoparticles assisted characterization of amine functionalized polystyrene multiwell plate and glass slide surfaces

et al. 2014

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We demonstrated citrate-capped gold nanoparticles assisted characterization of amine functionalized polystyrene plate and glass slide surfaces through AuNPs staining method. The effect of AuNPs concentration on the characterization of amine modified surfaces was also studied with different concentration of AuNPs (ratios 1.0-0.0). 3-Aminopropylyl triethoxy silane has been used as amine group source for the surface modification. The interactions of AuNPs on modified and unmodified surfaces were investigated using atomic force microscopy and the dispersibility, and the aggregation of AuNPs was analyzed using UV-visible spectrophotometer. Water contact angle measurement and X-ray photoelectron spectroscopy (XPS) were used to further confirmation of amine modified surfaces. The aggregation of AuNPs in modified multiwell plate leads to the color change from red to purple and they are found to be adsorped on the modified surfaces. Aggregation and adsorption of AuNPs on the modified surfaces through the electrostatic interactions and the hydrogen bonds were revealed by XPS analysis. Remarkable results were found even in the very low concentration of AuNPs (ratio 0.2). This AuNPs staining method is simple, cost-effective, less time consuming, and required very low concentration of AuNPs. These results can be read out through the naked eye without the help of sophisticated equipments.

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Section: Introductionmentioning

confidence: 99%

Gold nanoparticles assisted characterization of amine functionalized polystyrene multiwell plate and glass slide surfaces

et al. 2014

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“…[28] We co-immobilized amine functionalized PEG-maleimide (NH 2 -PEG-Mal, MW 3.4k Da) and PEG-hydroxyl (NH 2 -PEG-OH, MW 2k Da) to improve the nonfouling character of the surface and also cover the ‘defects’ on the polystyrene plate. [29] NH 2 -PEG-Mal groups were used for immobilization of dendrimer. Therefore, NH 2 -PEG-Mal grafting density reflects the dendrimer graft density on the surface.…”

Section: Resultsmentioning

confidence: 99%

Multifunctional Dendrimer‐Templated Antibody Presentation on Biosensor Surfaces for Improved Biomarker Detection

Han

Kannan

Wang

et al. 2010

Adv Funct Materials

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Dendrimers, with their well-defined globular shape and a high density of functional groups, are ideal nanoscale materials for templating sensor surfaces. This work exploits dendrimers as a versatile platform for capturing biomarkers with improved sensitivity and specificity. Synthesis, characterization, fabrication, and functional validation of the dendrimer-based assay platform are described. Bifunctional hydroxyl/thiol functionalized G4-polyamidoamine (PAMAM) dendrimer is synthesized and immobilized on to the polyethylene-glycol (PEG)-functionalized assay plate by coupling PEG-maleimide and dendrimer thiol groups. Simultaneously, part of the dendrimer thiol groups are converted to hydrazide functionalities. The resulting dendrimer-modified surface is coupled to the capture antibody in the Fc region of the oxidized antibody. This preserves the orientation flexibility of the antigen binding region (Fv) of the antibody. To validate the approach, the fabricated plates are further used as a solid phase for developing a sandwich type ELISA to detect IL-6 and IL-1β, important biomarkers for early stages of chorioamnionitis. The dendrimer-modified plate provides assays with significantly enhanced sensitivity, lower nonspecific adsorption, and a detection limit of 0.13 pg ml-1 for IL-6 luminol detection and 1.15 pg ml-1 for IL-1β TMB detection, which are significantly better than those for the traditional ELISA. The assays were validated in human serum samples from normal (non-pregnant) woman and pregnant women with pyelonephritis. The specificity and the improved sensitivity of the dendrimer-based capture strategy could have significant implications for the detection of a wide range of cytokines and biomarkers since the capture strategy could be applied to multiplex microbead assays, conductometric immunosensors and field effect biosensors.

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“…On the other hand, non specific protein adsorption to the plastic surfaces is a well known problem (Rebeski et al 1999). Therefore, prior to the construction of the microfluidic device, the Au-PS was firstly functionalized with SAM of 16-MHA (Fig.…”

Section: Antibody Binding Onto Polystyrene and Microfluidic Systemmentioning

confidence: 99%

Antibody immobilization on to polystyrene substrate—on-chip immunoassay for horse IgG based on fluorescence

Darain

Gan

Tjin

2009

Biomed Microdevices

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A simple microfluidic immunoassay card was developed based on polystyrene (PS) substrate for the detection of horse IgG, an inexpensive model analyte using fluorescence microscope. The primary antibody was captured onto the PS based on covalent bonding via a self-assembled monolayer (SAM) of thiol to pattern the surface chemistry on a gold-coated PS. The immunosensor chip layers were fabricated from sheets by CO(2) laser ablation. The functionalized PS surfaces after each step were characterized by contact angle measurement, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). After the antibody-antigen interaction as a sandwich immunoassay with a fluorescein isothiocyanate (FITC)-conjugated secondary antibody, the intensity of fluorescence was measured on-chip to determine the concentration of the target analyte. The present immunosensor chip showed a linear response range for horse IgG between 1 microg/ml and 80 microg/ml (r = 0.971, n = 3). The detection limit was found to be 0.71 microg/ml. The developed microfluidic system can be extended for various applications including medical diagnostics, microarray detection and observing protein-protein interactions.

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Identification of unacceptable background caused by non-specific protein adsorption to the plastic surface of 96-well immunoassay plates using a standardized enzyme-linked immunosorbent assay procedure

Cited by 51 publications

References 2 publications

Gold nanoparticles assisted characterization of amine functionalized polystyrene multiwell plate and glass slide surfaces

Gold nanoparticles assisted characterization of amine functionalized polystyrene multiwell plate and glass slide surfaces

Multifunctional Dendrimer‐Templated Antibody Presentation on Biosensor Surfaces for Improved Biomarker Detection

Antibody immobilization on to polystyrene substrate—on-chip immunoassay for horse IgG based on fluorescence

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