Please use this identifier to cite or link to this item: http://hdl.handle.net/1959.3/2221

Download PDF (Published version) (Adobe Acrobat PDF, -1 bytes)

Title

Oligonucleotide/poly(1-lysine) complexes attachment on poly(styrene/maleic acid) and poly(styrene/maleic anhydride) polymeric surfaces

Author(s)

Ivanova, Elena P.;  Pham, Duy K.;  Demyashev, Gregory M.;  Nicolau, Dan V.

Abstract

The immobilization efficiency of the complexes of oligonucleotide/poly(L-lysine) onto Poly(Styrene/Maleic Acid), PSMA, and Poly(Styrene/Maleic Anhydride), PSMAA, has been investigated using X-ray photoelectron spectroscopy and atomic force microscopy (AFM) in conjugation with fluorescence-based measurements of DNA attachment. A mono-molecularly thin layer of either electrostatically or covalently (via amide bond) coupled poly(L-lysine) (PL) allows the "switching" of the chemistry from a COOH-based to NH2-based one. The COOH-based chemistry has the advantage of a high yield of reaction but the disadvantage of a low surface concentration of DNA molecules (negative-negative electrostatic exclusion) whereas the NH2-based chemistry provides a higher surface concentration (positive-negative electrostatic attraction) but has a lower yield of covalent binding reaction. The immobilization efficiency of covalently coupled 26-mer oligonucleotides/poly(L-lysine) to polymeric surfaces was estimated as 0.3-0.5 x1012 molecules/mm2 for both polymeric surfaces studied. The electrostatic adsorption of poly(L-lysine)/oligonucleotides onto PSMA and functionalized PSMAA surfaces yielded 0.5 x 1011 and 0.1 x 1010 molecules/mm2, respectively. Although this mode of attachment is not "covalent binding" per se, the evidence is provided that this attachment is strong enough to withstand PCR cycles. The properties of these oligonucleotide/poly(L-lysine) complexes make them promising candidates for DNA-DNA hybridisation assays and PCR.

Publication type

Conference paper

Research centre

Swinburne University of Technology. Industrial Research Institute Swinburne

Source

Proceedings of SPIE: Biomedical Applications of Micro- and Nanoengineering, Melbourne, Victoria, Australia, 16-18 December 2002, Vol. 4937, pp. 23-33

Publication year

2002

Publisher

SPIE: International Society for Optical Engineering

Publisher URL

http://dx.doi.org/10.1117/12.471953

Copyright

Copyright © 2003 SPIE Society of Photo-Optical Instrumentation Engineers. This paper was originally published in Proceedings of SPIE (Vol. 4937), and is available from: http://dx.doi.org/10.1117/12.471953. The published version of the paper is reproduced here in accordance with the copyright policy of the publisher. One print or electronic copy may be made for personal use only. Systematic electronic or print reproduction and distribution, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper are prohibited.

Full text

Peer reviewed