Selection of DNA aptamers using atomic force microscopy

Atomic force microscopy (AFM) can detect the adhesion or affinity force between a sample surface and cantilever, dynamically. This feature is useful as a method for the selection of aptamers that bind to their targets with very high affinity. Therefore, we propose the Systematic Evolution of Ligands by an EXponential enrichment (SELEX) method using AFM to obtain aptamers that have a strong affinity for target molecules. In this study, thrombin was chosen as the target molecule, and an ‘AFM-SELEX’ cycle was performed. As a result, selected cycles were completed with only three rounds, and many of the obtained aptamers had a higher affinity to thrombin than the conventional thrombin aptamer. Moreover, one type of obtained aptamer had a high affinity to thrombin as well as the anti-thrombin antibody. AFM-SELEX is, therefore, considered to be an available method for the selection of DNA aptamers that have a high affinity for their target molecules.

[1]  Elisabeth Kremmer,et al.  Development and characterization of new rat monoclonal antibodies for procalcitonin , 2008, Analytical and bioanalytical chemistry.

[2]  R. Weiss,et al.  Binding of biotinylated DNA to streptavidin-coated polystyrene latex: effects of chain length and particle size. , 1996, Analytical biochemistry.

[3]  Zhaohu Li,et al.  Monoclonal antibody-based enzyme linked immunosorbent assay for the analysis of jasmonates in plants. , 2008, Journal of integrative plant biology.

[4]  L. Gold,et al.  Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. , 1990, Science.

[5]  Penmetcha K. R. Kumar,et al.  Selection of RNA aptamers against human influenza virus hemagglutinin using surface plasmon resonance. , 2005, Analytical biochemistry.

[6]  K. Kaneko,et al.  Screening of DNA Aptamer Against Mouse Prion Protein by Competitive Selection , 2007, Prion.

[7]  M Yarus,et al.  Diversity of oligonucleotide functions. , 1995, Annual review of biochemistry.

[8]  M. Hoylaerts,et al.  The application of enzyme kinetics to the determination of dissociation constants for antigen-antibody interactions in solution. , 1990, Journal of immunological methods.

[9]  J. Szostak,et al.  A DNA aptamer that binds adenosine and ATP. , 1995, Biochemistry.

[10]  T. Cech,et al.  Specific interaction between the self-splicing RNA of Tetrahymena and its guanosine substrate: implications for biological catalysis by RNA , 1984, Nature.

[11]  Itamar Willner,et al.  Following aptamer-thrombin binding by force measurements. , 2006, Analytical chemistry.

[12]  K. Dill,et al.  Determination of dissociation constant and concentration of an anti-DNA antibody by using the light-addressable potentiometric sensor. , 1996, Journal of biochemical and biophysical methods.

[13]  J. Szostak,et al.  In vitro selection of functional nucleic acids. , 1999, Annual review of biochemistry.

[14]  K. Ikebukuro,et al.  Aptameric enzyme subunit for homogeneous DNA sensing , 2007, Biotechnology Letters.

[15]  T. Whiteside CHAPTER 61 – Assays for cytokines , 2003 .

[16]  M. Gu,et al.  Electrochemical detection of 17β-estradiol using DNA aptamer immobilized gold electrode chip , 2007 .

[17]  Koji Sode,et al.  Selection of DNA aptamers against VEGF165 using a protein competitor and the aptamer blotting method , 2008, Biotechnology Letters.

[18]  Jijun Tang,et al.  In vitro selection of DNA aptamer against abrin toxin and aptamer-based abrin direct detection. , 2007, Biosensors & bioelectronics.

[19]  Hao Yan,et al.  Recognition imaging with a DNA aptamer. , 2006, Biophysical journal.

[20]  A. Kobayashi,et al.  In vitro selection of hematoporphyrin binding DNA aptamers. , 2000, Bioorganic & medicinal chemistry letters.

[21]  Man Bock Gu,et al.  ssDNA aptamers that selectively bind oxytetracycline. , 2008, Bioorganic & medicinal chemistry.

[22]  P. Soman,et al.  AFM measurements of interactions between the platelet integrin receptor GPIIbIIIa and fibrinogen. , 2009, Colloids and surfaces. B, Biointerfaces.

[23]  R. Stoltenburg,et al.  In vitro selection of DNA aptamers binding ethanolamine. , 2005, Biochemical and biophysical research communications.

[24]  E. Vermaas,et al.  Selection of single-stranded DNA molecules that bind and inhibit human thrombin , 1992, Nature.

[25]  Andrew D. Ellington,et al.  Nucleic Acid Selection and the Challenge of Combinatorial Chemistry. , 1997, Chemical reviews.

[26]  X Chris Le,et al.  Assays for cytokines using aptamers. , 2006, Methods.

[27]  J. Szostak,et al.  Isolation of high-affinity GTP aptamers from partially structured RNA libraries , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[28]  B. Nall,et al.  Measurement of the dissociation rate constant of antigen/antibody complexes in solution by enzyme-linked immunosorbent assay. , 1994, Journal of immunological methods.

[29]  Sonia Centi,et al.  Analytical performances of aptamer-based sensing for thrombin detection. , 2007, Analytical chemistry.

[30]  A. Kondo,et al.  Selection of a DNA aptamer that binds 8-OHdG using GMP-agarose. , 2009, Bioorganic & medicinal chemistry letters.

[31]  S. Otsuka,et al.  Development of glutathione‐coupled cantilever for the single‐molecule force measurement by scanning force microscopy , 2006, FEBS letters.