Journal
CARBOHYDRATE POLYMERS
Volume 131, Issue -, Pages 80-89Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carbpol.2015.05.043
Keywords
Chitosan; Pyrrolidinyl peptide nucleic acid; Quaternary ammonium group; MALDI-TOF mass spectrometry; DNA sequence analysis
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Funding
- Thailand Research Fund
- Office of Higher Education Commission [RTA5080004, RTA5280002]
- Chulalongkorn University [DPG5780002]
- Ratchadaphiseksomphot Endowment Fund of Chulalongkorn University [RE5560530126-AM]
- Thai Government Stimulus Package 2, under the Project for Establishment of Comprehensive Center for Innovative Food, Health Products and Agriculture [TKK2555]
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Quaternized chitosan particles are introduced as anion-exchanged captures to be used with a conformationally constrained pyrrolidinyl peptide nucleic acid (acpcPNA) and MALDI-TOF mass spectrometry for DNA sequence analysis. Methylated chitosan (MC) and methylated N-benzyl chitosan (MBzC) particles were obtained by heterogeneous chemical modification of ionically cross-linked chitosan particles via direct methylation and reductive benzylation/methylation, respectively. N,N,N-trimethylchitosan (TMC) and N-[(2-hydroxyl-3-trimethylammonium)propyl]chitosan chloride (HTACC) particles were prepared by ionic cross-linking of quaternized chitosan derivatives, homogeneously modified from chitosan, namely TMC and HTACC, respectively. The particles formed had a size in a sub-micrometer range and possessed positive charge. Investigation by MALDI-TOF mass spectrometry suggested that some quaternized particles in combination with acpcPNA were capable of detecting a single mismatched base out of 9-14 base DNA sequences. Potential application of this technique for the detection of wild-type and mutant K-ras DNA, a gene that mutation is associated with certain cancers, has also been demonstrated. (C) 2015 Elsevier Ltd. All rights reserved.
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