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

Abstract

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.

Bonding of thermoplastic microfluidics by using dry adhesive tape

“…28,29 Dry adhesive is also capable of multifunction integration 30 and reversible sealing. 31 Due to these advantages, dry adhesive bonding has been applied in various microuidic applications including droplet-based micro-uidics, 32 electrowetting microdevice, 33,34 chip-based electrophoresis, 35 electrochemical biosensing, 36 microarray immunoassay device, 37 and optical biochips. 38 Surface chemistry and biocompatibility is one of the critical considerations for dry adhesive bonding in microuidic applications.…”

Mitigated reactive oxygen species generation leads to an improvement of cell proliferation on poly[glycidyl methacrylate‐co‐poly(ethylene glycol) methacrylate] functionalized polydimethylsiloxane surfaces

“…The enthusiasm of the modification of PDMS will be enhanced with a bifunctional polymer brush that can reduce the hydrophobicity of PDMS while providing functional groups for covalently tethering protein without further activation or involving of chemical linkers, such as EDC/NHS and glutaraldehyde. Liu et al 26,27 have reported that poly(glycidyl methacrylate-co-poly(ethylene glycol) methacrylate (poly(GMA-copEGMA)) brush-functionalized glass surface can robustly capture target proteins and reduce nonspecific binding of unrelated biomolecules for the construction of sensitive immunoassay. The impact of poly(GMA-co-pEGMA) functionalization to cell adhesion and growth is poorly understood.…”

“…Liu et al have reported that poly(glycidyl methacrylate‐ co ‐poly(ethylene glycol) methacrylate (poly(GMA‐ co ‐pEGMA)) brush‐functionalized glass surface can robustly capture target proteins and reduce nonspecific binding of unrelated biomolecules for the construction of sensitive immunoassay. The impact of poly(GMA‐ co ‐pEGMA) functionalization to cell adhesion and growth is poorly understood.…”

Synthesis and self-assembly behavior of amphiphilic methyl α-D-glucopyranoside-centered copolymers

“…The epoxide groups can be also used for several subsequent couplings, i.e. biofunctionalization via the attachment of amine functionalized biological molecules, such as peptides, antibodies, and DNA [7,[19][20][21].…”