G‐rich sequence‐functionalized polystyrene microsphere‐based instantaneous derivatization for the chemiluminescent amplified detection of DNA

Abstract

Herein, we develop a novel chemiluminescence (CL) approach with high sensitivity and excellent selectivity, by taking advantage of magnetic beads as preconcentration carriers and polystyrene microspheres as an amplification platform. Briefly, a 'sandwich-type' detection strategy is employed in our design, which involves capture probe DNA immobilized on the surface of carboxyl-terminated magnetic beads and multiple biotinylated reporter DNA self-assembled on the surface of streptavidin-modified polystyrene microspheres. The reporter DNA includes a guanine nucleobase-rich (G-rich) sequence domain for the generation of light and an additional tethered nucleic acid domain complementary with the target DNA. The CL signal is obtained via a novel instantaneous derivatization reaction between a specific CL reagent and the guanine nucleo-bases rich in the target and reporter DNA. As a result, we demonstrate that this DNA assay is reproducible, stable, easy to use, and can sensitively detect femtomolar target DNA related to anthrax lethal factors with excellent differentiation ability for single-base mismatched sequences. Overall, this new CL protocol couples the high sensitivity of CL analysis with effective magnetic separation for discriminating against unwanted constituents such as mismatched sequences, and hence, offers great promise for DNA hybridization analysis.

Simultaneous detection of two lung cancer biomarkers using dual-color fluorescence quantum dots

“…Multiple biotinylated QDs self-assembled on the surface of streptavidinmodified polystyrene beads or carbon nanotubes could be envisioned to further extend the power of this protocol. [40][41][42][43] Second, antibody pairs for CEA and NSE need to be further optimized to avoid possible non-specific interaction and thus produce a reliable and robust multiplexed assay. Despite these challenges and limitations, the technique proposed here is universal since the same method can be used for multianalyte quantification of any sample.…”

Microbiological Sensing Technologies: A Review

“…Immobilization strategies differ mainly in the natures of the functionalized surface and the bio-reporter [ 22 ]. For different applications, the bio-reporter can be immobilized on different types of functionalized slide surfaces, including glass [ 23 ], silicon [ 24 ], PDMS (polydimethylsiloxane) [ 25 ], COC (cyclic olefin copolymer) [ 26 ], PS (polystyrene) [ 27 ], PMMA (polymethyl-methacrylate) [ 28 ], and metal films [ 29 ]. For the majority of these slides, the surface is first covered with a layer of a silanizing agent, such as siorganofunctional alkoxysilane molecules, which provide suitable functional residues for anchoring to functional groups (like -OH and -NH 2 ) of proteins [ 30 ].…”

A novel method to prepare magnetic polymer‐based anion exchangers and their application

“…Magnetic polymer microspheres have a wide range of biological applications, including enzyme immobilization, nucleic acid sequencing, and separation of biomolecules (DNA, RNA, and protein) and cells . The use of magnetic microspheres has the advantage of allowing quick and one‐step purification of biological substances without centrifugation.…”