Surface characterization of oligonucleotides immobilized on polymer surfaces

Duy K. Pham; Elena P Ivanova; Jonathan P. Wright; Piotr A. Grodzinski; Ralf Lenigk; Dan V. Nicolau

doi:10.1117/12.471946

14 November 2002 Surface characterization of oligonucleotides immobilized on polymer surfaces

Duy K. Pham, Elena P Ivanova, Jonathan P. Wright, Piotr A. Grodzinski, Ralf Lenigk, Dan V. Nicolau

Proceedings Volume 4937, Biomedical Applications of Micro- and Nanoengineering; (2002) https://doi.org/10.1117/12.471946
Event: SPIE's International Symposium on Smart Materials, Nano-, and Micro- Smart Systems, 2002, Melbourne, Australia

Abstract

The immobilization and hybridization of amino-terminated oligonucleotide strands to cyclo-olefin-copolymer (COC) and polycarbonate (PC) surfaces have been investigated for potential application in micro-PCR devices. The oligonucleotides were covalently bound to the plasma-treated COC and PC surfaces via an N-hydroxy-sulfosuccinimide (NHSS) intermediate. Analysis by AFM showed that the oligonucleotides were present on the surfaces as lumps, and that the size, both vertically and laterally, of these lumps on the COC surface was larger compared to the PC surface. The immobilization efficiency of the former was also higher (15.8 x 10¹² molecules / cm²) compared to the latter (3.3 x 10¹² molecules / cm²). The higher efficiency of the COC surface is attributed to the more effective NHSS-functionalization and its higher surface roughness. Subsequent hybridization doubled the height of the lumps, while the lateral dimensions remained essentially unchanged. This is explained in terms of organization of the long probe strands used on the surface as flexible, coil-like polymer chains, which allow the complementary oligonucleotides to bind and increase the height of the lumps. The AFM frictional images showed that the hybridization had the effect of reversing hydrophilicity of the oligonucleotide lumps from being more hydrophilic to more hydrophobic, consistent with the hydrophilic bases of the probe strands being shielded as a result of hybridization.

Citation Download Citation

Duy K. Pham, Elena P Ivanova, Jonathan P. Wright, Piotr A. Grodzinski, Ralf Lenigk, and Dan V. Nicolau "Surface characterization of oligonucleotides immobilized on polymer surfaces", Proc. SPIE 4937, Biomedical Applications of Micro- and Nanoengineering, (14 November 2002); https://doi.org/10.1117/12.471946

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