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Filter paper grafted with epoxide-based copolymer brushes for activation-free peptide nucleic acid conjugation and its application for colorimetric DNA detection
- Source :
- Colloids and Surfaces B: Biointerfaces. 173:851-859
- Publication Year :
- 2019
- Publisher :
- Elsevier BV, 2019.
-
Abstract
- Epoxide-bearing filter paper was first prepared by surface-initiated reversible addition-fragmentation chain transfer (RAFT) copolymerization of glycidyl methacrylate (GMA) and poly(ethylene glycol)methacrylate (PEGMA). Without the need for activation step, the capture peptide nucleic acid (PNA) probes carrying a C-terminal lysine modification can be directly immobilized on the surface-grafted poly[glycidyl methacrylate-ran-poly(ethylene glycol)methacrylate] (P(GMA-ran-PEGMA)) through ring-opening of epoxide groups in the GMA repeating units by amino groups in the PNA’s structure. The success of P(GMA-ran-PEGMA) grafting on the filter paper and subsequent PNA immobilization was confirmed by fluorescence microscopy, Fourier transform-infrared spectroscopy and X-ray photoelectron spectroscopy. Colorimetric detection with signal amplification upon DNA hybridization relies on sandwich-hybridization assay employing another biotinylated PNA strand as a reporter probe together with streptavidin-horseradish peroxidase conjugate (SA-HRP) and o-phenylenediamine (OPD) substrate. It was found that increasing ionic strength during the DNA hybridization step by addition of NaCl can increase the signal intensity, which can be visualized by naked eye. The sensing platform showed the best performance in preventing non-specific adsorption from the non-complementary DNA and discriminating between complementary and single-mismatched targets of at least 50 fmol without the requirement for stringent hybridization or washing condition. This superior ability to suppress non-specific adsorption of non-target DNA as well as other non-DNA components may be explained as a result of hydrophilic PEGMA repeating units in the surface-grafted copolymer.
- Subjects :
- Peptide Nucleic Acids
Glycidyl methacrylate
Magnetic Resonance Spectroscopy
Polymers
Surface Properties
Epoxide
02 engineering and technology
Phenylenediamines
Methacrylate
01 natural sciences
Polyethylene Glycols
Polymerization
chemistry.chemical_compound
Colloid and Surface Chemistry
HLA Antigens
Spectroscopy, Fourier Transform Infrared
0103 physical sciences
Copolymer
Humans
Physical and Theoretical Chemistry
Alleles
Horseradish Peroxidase
010304 chemical physics
Peptide nucleic acid
Filter paper
Nucleic Acid Hybridization
Chain transfer
DNA
Surfaces and Interfaces
General Medicine
021001 nanoscience & nanotechnology
Combinatorial chemistry
chemistry
Epoxy Compounds
Methacrylates
Colorimetry
Adsorption
0210 nano-technology
Ethylene glycol
Filtration
Software
Biotechnology
Subjects
Details
- ISSN :
- 09277765
- Volume :
- 173
- Database :
- OpenAIRE
- Journal :
- Colloids and Surfaces B: Biointerfaces
- Accession number :
- edsair.doi.dedup.....5b4a2c460a57431660bdf68837b41b52