Three-Dimensional Flow-Through Protein Platform

Lieshout, R. Van; Domburg, T. van; Saalmink, Milan; Verbeek, R.; Wimberger-Friedl, R.; Dieijen-Visser, Marja van; Punyadeera, Chamindie

doi:10.1021/ac801244d

Cited by 15 publications

(14 citation statements)

References 18 publications

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“…; Figure 1b); hence, the incubation time for the antigen-antibody reaction corresponded to just the permeation time, which was 1/20 shorten than that of conventional ELISA systems (37 1C 1 h) and (3) this rapid and precise centrifugal permeation system can be applied to various molecular diagnostic biosensors because of its simple, reliable and disposable process, which improves other approaches for permeation detection in existence. [13][14][15][16][17] Moreover, we considered that this permeation detection system can achieve the separation and detection of antigen molecules from whole blood simultaneously, just when the molecular cutoff of the filter is appropriate to the molecular weight of the antigen.…”

Section: Introductionmentioning

confidence: 99%

Polyelectrolyte multilayers-modified membrane filter for rapid immunoassay: protein condensation by centrifugal permeation

Shen

Watanabe

Akashi

2010

Polym J

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We demonstrated that positive polyelectrolyte multilayers (PEMs) have great potential for conventional enzyme-linked immunosorbent assay (ELISA) systems because of the induction protein enrichment on their surface. In this study, we developed a novel simple molecular detection system using a PEMs-modified cellulose acetate (CA) membrane filter (PEMs-CA), which achieved rapid detection without compromising sensitivity as compared with conventional ELISA systems. This rapid detection was carried out by the centrifugal permeation of the antigen solution, thus allowing the local condensation of the antigen molecule in the proximity of the primary antibody-enriched PEMs-CA membrane, which overcame molecular diffusion as the time-limiting factor as compared with conventional ELISA systems. Hence, on the permeation system, the incubation time required for the antigen-antibody reaction corresponded to just the permeation time, which was 1/20 shorten than the conventional ELISA system under optimized conditions for the centrifugal forces. Moreover, the calibration curve of PEMs-CA had wide range of concentration from 0.02 to 5 lg ml À1 and larger change in signal as compared with the bare CA membrane. We concluded that this centrifugal permeation system should be further developed for rapid, precise and simple systems in various immunosensors using PEMs-modified membrane filters. Keywords: centrifugal condensation; ELISA; membrane filter; permeation detection; polyelectrolyte multilayers INTRODUCTIONSince immunoassays were first developed in the 1950s, antibodies and other affinity reagents have been developed to improve these assays in terms of specificity and sensitivity. 1 Moreover, the fundamental technique for using various precise immunosensors, electrochemical, optical and microgravimetric immunosensors has evolved significantly from the early devices that appeared in the late 1980s to the more sophisticated, integrated systems developed more recently. 2 Economic pressures in the health-care system mean that attempts have long been made to reconcile the elements of the tension triangle: 'fast-goodcheap' . Thus far, however, it has only been possible to satisfy only two of these elements. 3,4 Disposable immunotests (qualitative and quantitative), such as enzyme-linked immunosorbent assay (ELISA), retain the gold standard for protein detection and quantification. However, long incubation times are required to reach complete antigen-antibody reactions due to the long diffusion times of the antigen molecules toward the antibody as the rate-limiting step in these systems, resulting in slow binding kinetics and compromised assay sensitivity and dynamic range. [5][6][7][8] In this study, we developed a rapid, precise and simple antigen permeation immunoassay based on the positive polyelectrolyte multilayers (PEMs)-modified membrane filter (cellulose acetate,

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Section: Introductionmentioning

confidence: 99%

Polyelectrolyte multilayers-modified membrane filter for rapid immunoassay: protein condensation by centrifugal permeation

Shen

Watanabe

Akashi

2010

Polym J

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“…An optical flow-through assay using a fibrous filter, has exposed the capture surface to the entire analyte solution to avoid mass transport limitations [5], but it was an ex-situ end point assay with complicated steps. Recently, free standing 200 nm pore AAO membranes, when functionalized with an epoxysilane, allowed analytes to flow-through less than 100 nm from the assay surface, breaking mass transport limitations and effectively targeting their delivery [6]. In that work, polarimetry (Fig 1) was used to take the preliminary real-time biosensing results.…”

Section: Introductionmentioning

confidence: 99%

A real time immunoassay in alumina membranes

Álvarez

Solá

Cretich

et al. 2014

IEEE SENSORS 2014 Proceedings

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Abstract-To date, photonic biosensing with porous membranes has produced slow responses and long sensing times, due to the narrow (less than 100 nm) closed end pores of the membranes used. Recently, polarimetry was used to demonstrate analyte flow through, and real time biosensing in, free-standing porous alumina membranes. Here, we demonstrate how an improved functionalization technology, has for the first time enabled a real-time immunoassay within a porous membrane with a total assay time below one hour. With the new approach, we show a noise floor for individual biosensing measurements of 3.7 ng/ml (25 pM), and a bulk refractive index detection limit of 5×10 -6 RIU, with a standard deviation of less than 5%. The membranes, with their 200 nm pore diameter enabling targeted delivering of analytes to bioreceptors immobilized on the pore walls, therefore provide a route towards rapid and low cost realtime opto-fluidic biosensors for small sample volumes.

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“…The subsequent activation via oxidation or homobifunctional crosslinking could also result in low activation efficiency and unstable intermediates such as schiff's base (Bai et al 2006;Yu et al 2009b). Rigid porous matrixes including polymer monoliths (Jain et al 2004;) and organic/inorganic membranes (Dai et al 2006;Fu et al 2008;van Lieshout et al 2009) provide larger surface areas for high protein loading capacity to improve detection sensitivity. Nevertheless, in a flow operation, only the probes on the outside surface of the porous matrix could efficiently interact with the targets, but those in the inner pores are accessed with difficulty by the targets in such a short contact time.…”

Section: Introductionmentioning

confidence: 99%

Highly sensitive poly[glycidyl methacrylate-co-poly(ethylene glycol) methacrylate] brush-based flow-through microarray immunoassay device

Liu

Wang

et al. 2011

Biomed Microdevices

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Flow-through immunoassay is an attractive method for fast, inexpensive and high-throughput protein analyses. However, its practical application is limited by low sensitivity. In this work, a highly sensitive flow-through microarray immunoassay device is developed, in which a poly[glycidyl methacrylate-co-poly(ethylene glycol) methacrylate] (P(GMA-co-PEGMA)) brush as a flexible matrix is uniformly coated on a glass slide through a purge-free surface-initiated atom transfer radical polymerization (SI-ATRP) to immobilize capture proteins for a larger loading capacity and higher bioactivity while allowing easy target access to the brush-attached probes for efficient antibody-antigen (Ab-Ag) bindings. The integrated device is then constructed by simply laminating the protein-arrayed slide onto a ready-for-bonding double-sided adhesive tape-attached poly(methyl methacrylate) (PMMA) microfluidic structure. As a demonstration, a parallel microarray panel is designed to perform flow-through immunoassays for good detection flexibility and simultaneous analysis of various samples. The limit of detection (LOD) of 1-10 pg/mL for detected target proteins is achieved, which is one to two orders better than those of reported flow-through immunoassays. The device also demonstrates significantly reduced total assay time over the static microarray immunoassay. The rapid and sensitive detection can be mainly ascribed to the P(GMA-co-PEGMA) brushed substrate, of which both the hydrophilicity from its PEG component and the binding capability from its GMA moiety result in higher protein loading capacity, lower nonspecific adsorption, and higher Ab-Ag binding efficiency. The integrated microfluidic device was further used to detect an important cancer biomarker carcinoembryonic antigen (CEA) in serum and achieved a LOD of 10 pg/mL, demonstrating its great potential for clinical applications.

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Three-Dimensional Flow-Through Protein Platform

Cited by 15 publications

References 18 publications

Polyelectrolyte multilayers-modified membrane filter for rapid immunoassay: protein condensation by centrifugal permeation

Polyelectrolyte multilayers-modified membrane filter for rapid immunoassay: protein condensation by centrifugal permeation

A real time immunoassay in alumina membranes

Highly sensitive poly[glycidyl methacrylate-co-poly(ethylene glycol) methacrylate] brush-based flow-through microarray immunoassay device

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