Development of a novel bead-based 96-well filtration plate competitive immunoassay for the detection of Gentamycin

Ho, Tien Yu; Chan, Chia Chung; Chan, King Ho; Wang, Yu-Chieh; lin, Jing; Chang, Chun–Ju; Chen, Chien Sheng

doi:10.1016/j.bios.2013.04.027

Cited by 25 publications

(13 citation statements)

References 24 publications

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“…The preparation of the sulforhodamine B sodium (SRB)-encapsulated liposome was described in detail in previous publication. 29 The working concentration were 540 μg mL −1 dissolved in PBS. The substrates were first immersed in PBS for 30 min to achieve equilibrium before liposome adhesion.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Modification of Silicone Elastomer with Zwitterionic Silane for Durable Antifouling Properties

et al. 2014

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Biofouling on medical devices generally causes adverse complications, such as thrombosis, infection, and pathogenic calcification. Silicone is a widely used material for medical applications. Its surface modification typically encounters undesirable "hydrophobic recovery", leading to deterioration of surface engineering. In this study, we developed a stable superhydrophilic zwitterionic interface on polydimethylsiloxane (PDMS) elastomer by covalent silanization of sulfobetaine silane (SBSi) to resist nonspecific adsorption of bacteria, proteins, and lipids. SBSi is a zwitterionic organosilane assembly, enabling resisting surface reconstruction by forming a cross-linked network and polar segregation. Surface elemental composition was confirmed by X-ray photoelectron spectroscopy (XPS), and the long-term stability of modification was accessed using a contact angle goniometer. The biofouling tests were carried out by exposing substrates to bacterial, protein, and lipid solutions, revealing the excellent bioinertness of SBSi-tailored PDMS, even after 30 day storage in ambient. For the real-world application, we modified commercially available silicone hydrogel contact lenses with developed zwitterionic silane, presenting its antibacterial adhesion property. Moreover, the cytotoxicity of SBSi was accessed with NIH-3T3 fibroblast by the MTT assay, showing negligible cytotoxicity up to a concentration of 5 mM. Consequently, the strategy of surface engineering in this work can effectively retard the "hydrophobic recovery" occurrence and can be applied to other silicone-based medical devices in a facile way.

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Section: ■ Materials and Methodsmentioning

confidence: 99%

Modification of Silicone Elastomer with Zwitterionic Silane for Durable Antifouling Properties

et al. 2014

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“…Flow-through direct immunoassays are performed into wells of a 96-well filter plates that have a membrane (Durapore PVDF, 0.45 um pore size) in the bottom of each microwell. Filter well plates are typically used for analyte purifications (plasmid isolation, DNA, and PCR products purification) and receptor/ligand binding assays but they have been also used in analytical assays such as ELISpot assays, bacterial colony count assays, ELISA assays for detection of antibodies, , and bead-based immunoassays . To the best of our knowledge, they have not been used in assays for the detection of bacteria.…”

Section: Resultsmentioning

confidence: 99%

“…Filter well plates are typically used for analyte purifications (plasmid isolation, DNA, and PCR products purification) and receptor/ligand binding assays but they have been also used in analytical assays such as ELISpot assays, 25 bacterial colony count assays, 26 ELISA assays for detection of antibodies, 6,27 and bead-based immunoassays. 28 To the best of our knowledge, they have not been used in assays for the detection of bacteria. Each well of a filter plate can hold 300 μL of a solution, but higher volumes can be sampled as the wells can be drained from the bottom.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

High-Throughput Flow-Through Direct Immunoassays for Targeted Bacteria Detection

et al. 2021

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Regulatory authorities require analytical methods for bacteria detection to analyze large sample volumes (typically 100 mL). Currently only the Membrane Filtration and the Most Probable Number assays analyze such large volumes, while other assays for bacteria detection (ELISA, lateral flow assays, etc.) typically analyze volumes 1000 times smaller. This study describes flow-through direct immunoassays (FTDI), a new methodology for the targeted detection of bacteria in liquid samples of theoretically any volume. Flow-through direct immunoassays are performed in fluid-permeable microwells (e.g., wells of a filter well plate) that have a membrane on their bottom where the bacteria are trapped before their detection using a direct immunoassay. Two versions of FTDI assays for the detection of E. coli in 10 mL of sample were developed. A rapid FTDI assay that can be completed in less than 2.5 h can detect E. coli bacteria in levels down to 17 CFU/mL, and an ultrasensitive FTDI assay that employs an additional bacteria culturing step to boost the sensitivity can detect E. coli bacteria in levels lower than 1 CFU/mL in less than 5.5 h. All the steps of the assays, including the immunoassay steps, the culturing step, and the analytical signal measurement step are performed inside the well plate to decrease the chance of contamination and ensure a safe, easy process for the user. The assays were assessed and validated in tap water, river water, and apple juice samples, and the results suggests that the assays are robust, precise, and accurate. When the assays are performed in 96-well filter plates, a filter well plate vacuum manifold and a multichannel peristaltic pump are also used, so multiple samples can be analyzed in parallel to allow highthroughput analysis of samples.

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“…The bead-based ELISA originates from sandwich ELISA and substitutes the flat plastic surface supports with magnetic, polymeric beads, or microspheres. Due to the mobility of the particles, bead-based ELISA can be applied in more detection systems. − Traditionally, the detection of the enzyme labels was performed by optical assays based on chromogenic and fluorogenic organic substrates, which change absorption or emission spectra in the course of enzymatic reactions. However, most of the organic dyes currently in use for bead-based ELISA are not very resistant against decomposition under the action of light in the presence of oxygen, hydrogen peroxide, and water in the assay mixture resulting in bleaching.…”

Section: Introductionmentioning

confidence: 99%

Microbead QD-ELISA: Microbead ELISA Using Biocatalytic Formation of Quantum Dots for Ultra High Sensitive Optical and Electrochemical Detection

Grinyte

Barroso

Möller

et al. 2016

ACS Appl. Mater. Interfaces

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Electrochemical detection strategies employing semiconductor quantum dots (QDs) open up new opportunities for highly sensitive detection of biological targets. We designed a new assay based on microbead linked enzymatic generation of CdS QDs (Microbead QD-ELISA) and employed it in optical and electrochemical affinity assays for the cancer biomarker superoxide dismutase 2 (SOD2). Biotinylated antibodies against SOD2 were immobilized on the surface of polyvinyl chloride microbeads bearing streptavidin. In order to prevent any non-specific adsorption the microbeads were further blocked with bovine serum albumin. The analyte, SOD2 was captured on microbeads and labeled with alkaline phosphatase-conjugated antibody linked with mouse antibody against SOD2. Hydrolysis of para-nitrophenylphosphate by immobilized alkaline phosphatase triggered the rapid formation of phosphate-stabilized CdS QDs on the surface of microbeads. The resulting semiconductor nanoparticles were detected by fluorescence spectroscopy, microscopy, and square-wave voltammetry (SWV). The electrochemical assay based on the detection with square-wave voltammograms of Cd ions originating from immobilized CdS QDs showed linearity up to 45 ng mL, and the limit of SOD2 detection equal to 0.44 ng mL (1.96 × 10 M). This detection limit is lower by 2 orders of magnitude in comparison with that of other previously published assays for superoxide dismutase. The electrochemical assay was validated with HepG2 (Human hepatocellular carcinoma) cell lysate containing SOD2.

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Development of a novel bead-based 96-well filtration plate competitive immunoassay for the detection of Gentamycin

Cited by 25 publications

References 24 publications

Modification of Silicone Elastomer with Zwitterionic Silane for Durable Antifouling Properties

Modification of Silicone Elastomer with Zwitterionic Silane for Durable Antifouling Properties

High-Throughput Flow-Through Direct Immunoassays for Targeted Bacteria Detection

Microbead QD-ELISA: Microbead ELISA Using Biocatalytic Formation of Quantum Dots for Ultra High Sensitive Optical and Electrochemical Detection

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