Journal
SENSORS AND ACTUATORS B-CHEMICAL
Volume 137, Issue 1, Pages 215-221Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2008.11.038
Keywords
Peptide nucleic acid; DNA; QCM; Self-assembled monolayer; Biosensor; Hybridization
Funding
- Ratchadapiseksompoj Endowment Fund
- National Research Council of Thailand (NRCT)
- Nanopolymer Project [GOR-SOR-SOR 52/2547, 12/2548]
- National Nanotechnology Center [NN-B-22-b6-10-49-17]
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Two novel thiolated pyrrolidinyl peptide nucleic acids (HS-PNA) carrying D-prolyl-2-aminocyclopentane-carboxylic acid (ACPC) backbones were synthesized and directly immobilized on gold-coated quartz crystals by self-assembled monolayer (SAM) formation (via S atom linkage) for the detection of DNA hybridization. The amount of immobilized HS-PNA was determined by quartz crystal microbalance (QCM), whilst the surface functionality and wettability of the gold surfaces before and after immobilization of the HS-PNA were characterized by reflection absorption infrared spectroscopy (RAIRS) and water contact angle analysis, respectively. Three different procedures for immobilization of HS-PNA were investigated; (I) HS-PNA without blocking thiol, (II) HS-PNA with subsequent blocking thiol and (III) co-immobilization with mixed HS-PNA and blocking thiol. The amount of immobilized HS-PNA on the gold electrode increased asymptotically with the HS-PNA concentration. The sequential blocking step procedure II) was critical to the success of subsequent DNA hybridization. For hybridization, 0.5 mN phosphate buffer at pH 7.0 with no added NaCl Was found to be optimal. Under these conditions, discrimination between complementary and single or multiple base mismatched DNA targets was achieved Using a combination of pyrrolidinyl HS-PNA with QCM. (C) 2008 Elsevier B.V. All rights reserved.
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