PNA-PEG Modified Silicon Platforms as Functional Bio-Interfaces for Applications in DNA Microarrays and Biosensors

摘要

The synthesis and characterization of two types of silicon-based biofunctional interfaces are reported;each interface bonds a dense layer of poly(ethylene glycol)(PEG")and peptide nucleic acid(PNA)probes.Phosphonate self-assembled monolayers were derivatized with PNA using a maleimido-terminated PEG_(45).Similarly,siloxane monolayers were functionalized with PNA using a maleimido-terminated PEG_(45)spacer and were subsequently modified with a shorter methoxy-terminated PEG_(12)("back-filling").The long PEG_(45)spacer was used to distance the PNA probe from the surface and to minimize undesirable nonspecific adsorption of DNA analyte.The short PEG_(12)"back-filler"was used to provide additional passivation of the surface against nonspecific DNA adsorption.X-ray photoelectron spectroscopic(XPS)analysis near the C 1s and N 1s ionization edges was done to characterize chemical groups formed in the near-surface region,which confirmed binding of PEG and PNA to the phosphonate and silane films.XPS also indicated that additional PEG chains were tethered to the surface during the backfilling process.Fluorescence hybridization experiments were carried out with complementary and noncDNA strands;both phosphonate and siloxane biofunctional surfaces were effective for hybridization of cDNA strands and significantly reduced nonspecific adsorption of the analyte.Spatial patterns were prepared by polydimethylsiloxane(PDMS)micromolding on the PNA-functionalized surfaces;selective hybridization of fluorescently labeled DNA was shown at the PNA functionalized regions,and physisorption at the probe-less PEG-functionalized regions was dramatically reduced.These results show that PNA-PEG derivatized phosphonate monolayers hold promise for the smooth integration of device surface chemistry with semiconductor technology for the fabrication of DNA biosensors.In addition,our results confirm that PNA-PEG derivatized self-assembled carboxyalkylsiloxane films are promising substrates for DNA microarray applications.