X-ray photoelectron spectroscopy and infrared spectroscopy study of maleimide-activated supports for immobilization of oligodeoxyribonucleotides.

Surface-tethered nucleic acids are widely applied in solid-phase assays in which complementary strands must be detected against a complex mixture of other sequences. In response to such needs, numerous methods have been developed for immobilizing nucleic acids on solid supports. Often, detailed analysis of associated chemical transformations and of potential side reactions is difficult to obtain. Combined use of planar and high surface area powder supports allows characterization using surface as well as bulk diagnostic techniques. This approach is followed in the present study in which X-ray photoelectron spectroscopy (XPS), transmission infrared spectroscopy (FTIR) and reactivity titrations are used to investigate siliceous supports modified with an aminosilane precursor followed by a maleimide-bearing crosslinker for attachment of nucleic acids. The supports retain maleimide activity for approximately a day when stored under buffer, but deactivation is accelerated under basic conditions or by incomplete conversion of the precursor aminosilane monolayer. Reactions involving the olefinic bond of the imide as well as its carbonyl groups are observed and analyzed. Attachment of sulfhydryl-terminated oligodeoxyribonucleotides is highly site specific, and immobilized strands exhibit excellent hybridization activity. Quantitative use of XPS for label-free determination of DNA coverage based on calibration against reference materials is also described.

[1]  R. Georgiadis,et al.  The effect of surface probe density on DNA hybridization. , 2001, Nucleic acids research.

[2]  P. Voort,et al.  Characterization and Chemical Modification of the Silica Surface , 1995 .

[3]  R. Georgiadis,et al.  Kinetic Control of Hybridization in Surface Immobilized DNA Monolayer Films , 2000 .

[4]  C R Cantor,et al.  High-density, covalent attachment of DNA to silicon wafers for analysis by maldi-tof mass spectrometry. , 1997, Analytical chemistry.

[5]  S. Matsui,et al.  Hydrolysis of some N-alkylmaleimides , 1979 .

[6]  J. H. Scofield,et al.  Hartree-Slater subshell photoionization cross-sections at 1254 and 1487 eV , 1976 .

[7]  S. Balasubramanian,et al.  Probing DNA Surface Attachment and Local Environment Using Single Molecule Spectroscopy , 2001 .

[8]  F. Blum,et al.  Molecular Mobility of the Interface in a Model Polymer Composite: A NMR Study , 1992 .

[9]  Yoshio Okahata,et al.  Hybridization of nucleic acids immobilized on a quartz crystal microbalance , 1992 .

[10]  J Wang,et al.  From DNA biosensors to gene chips. , 2000, Nucleic acids research.

[11]  A. Steel,et al.  Electrochemical quantitation of DNA immobilized on gold. , 1998, Analytical chemistry.

[12]  O. Blumenfeld,et al.  Reactions of N-ethylmaleimide with peptides and amino acids. , 1964, The Biochemical journal.

[13]  T. M. Herne,et al.  Characterization of DNA Probes Immobilized on Gold Surfaces , 1997 .

[14]  R. Levicky,et al.  Preparation of End-Tethered DNA Monolayers on Siliceous Surfaces Using Heterobifunctional Cross-Linkers , 2003 .

[15]  White,et al.  An Infrared Study of the Amine-Catalyzed Reaction of Methoxymethylsilanes with Silica. , 2000, Journal of colloid and interface science.

[16]  E. Southern,et al.  Sequence variation in genes and genomic DNA: methods for large-scale analysis. , 2000, Annual review of genomics and human genetics.

[17]  Chang Ok Kim,et al.  Characteristics of DNA Microarrays Fabricated on Various Aminosilane Layers , 2002 .

[18]  p‐Maleimidophenyl Isocyanate: A Novel Heterobifunctional Linker for Hydroxyl to Thiol Coupling. , 1995 .

[19]  Joseph Wang.  SURVEY AND SUMMARY From DNA biosensors to gene chips , 2000 .

[20]  E. Lukhtanov,et al.  Attachment of benzaldehyde-modified oligodeoxynucleotide probes to semicarbazide-coated glass. , 2001, Nucleic acids research.

[21]  Charles S. Fadley,et al.  Angle-resolved x-ray photoelectron spectroscopy , 1984 .

[22]  C. Powell,et al.  The NIST Electron Effective-Attenuation-Length Database , 2002 .

[23]  H. Hörhold,et al.  Investigation of quantitative SiOH determination by the silane treatment of disperse silica , 1988 .

[24]  S. Satija,et al.  Using Self-Assembly To Control the Structure of DNA Monolayers on Gold: A Neutron Reflectivity Study , 1998 .

[25]  Y. Rogers,et al.  Immobilization of oligonucleotides onto a glass support via disulfide bonds: A method for preparation of DNA microarrays. , 1999, Analytical biochemistry.

[26]  Elizabeth M. Boon,et al.  Single-base mismatch detection based on charge transduction through DNA. , 1999, Nucleic acids research.

[27]  N. Yamamoto,et al.  Microarray fabrication with covalent attachment of DNA using Bubble Jet technology , 2000, Nature Biotechnology.

[28]  Gil U. Lee,et al.  Covalent attachment of synthetic DNA to self-assembled monolayer films. , 1996, Nucleic acids research.

[29]  N. McIntyre,et al.  ESCA spectra and molecular charge distributions for some pyrimidine and purine bases , 1978 .

[30]  G. Turcatti,et al.  Solid phase DNA amplification: characterisation of primer attachment and amplification mechanisms. , 2000, Nucleic acids research.

[31]  C. Niemeyer,et al.  Dendrimer‐Activated Solid Supports for Nucleic Acid and Protein Microarrays , 2001, Chembiochem : a European journal of chemical biology.

[32]  A. Horgan,et al.  Reaction of N-phenyl maleimide with aminosilane monolayers. , 2004, Colloids and surfaces. B, Biointerfaces.

[33]  S. Mason,et al.  Fourier Transform Raman and infrared spectroscopy of N-phenylmaleimide and methylene dianiline bismaleimide , 1990 .

[34]  G. Socrates,et al.  Infrared Characteristic Group Frequencies , 1980 .

[35]  M. McGovern,et al.  High surface density immobilization of oligonucleotide on silicon. , 2001, The Analyst.

[36]  B. Weimer,et al.  Optimizing the immobilization of single-stranded DNA onto glass beads. , 2001, Journal of biochemical and biophysical methods.

[37]  D. Castner,et al.  Quantitative X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry characterization of the components in DNA. , 2004, Analytical chemistry.

[38]  Wolfgang Knoll,et al.  Oligonucleotide hybridization observed by surface plasmon optical techniques , 1995 .

[39]  J. Crivello.  Polyaspartimides: Condensation of aromatic diamines and bismaleimide compounds , 1973 .

[40]  D. Clark,et al.  Molecular core binding energies; carbon and nitrogen 1s levels for adenine , 1970 .

[41]  D. Leyden,et al.  Ammonia-Catalyzed Silylation Reactions of Cab-O-Sil with Methoxymethylsilanes. , 1988 .

[42]  D. Clark,et al.  Molecular core binding energies; carbon and nitrogen 1s levels for cytosine and thymine , 1970 .