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

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

doi:10.1117/12.471953

14 November 2002 Oligonucleotide/poly(l-lysine) complexes attachment on poly(styrene/maleic acid) and poly(styrene/maleic anhydride) polymeric surfaces

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

Author Affiliations +

Proceedings Volume 4937, Biomedical Applications of Micro- and Nanoengineering; (2002) https://doi.org/10.1117/12.471953
Event: SPIE's International Symposium on Smart Materials, Nano-, and Micro- Smart Systems, 2002, Melbourne, Australia

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.

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Elena P. Ivanova, Duy K. Pham, Gregory M. Demyashev, and Dan V. Nicolau "Oligonucleotide/poly(l-lysine) complexes attachment on poly(styrene/maleic acid) and poly(styrene/maleic anhydride) polymeric surfaces", Proc. SPIE 4937, Biomedical Applications of Micro- and Nanoengineering, (14 November 2002); https://doi.org/10.1117/12.471953

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