Efficient selection of glycoprotein-binding DNA aptamers via boronate affinity monolithic capillary.

Systematic evolution of ligands by exponential enrichment (SELEX) is the workhorse method for selecting aptamers that are capable of binding target molecules from a random oligonucleic acid library. However, conventional SELEX methods are associated with apparent drawbacks including labor-intensive, time-consuming, large reagent consumption and strong nonspecific binding with separation media. Herein, we report a boronate affinity monolithic capillary-based SELEX approach for rapid selection of high-specificity glycoprotein-binding DNA aptamers. Boronate affinity monolithic capillary is an advanced functional material appeared in recent years, which allows for facile capture/release of glycoproteins in a pH-switchable fashion. By using boronate affinity monolithic capillary as a platform for target immobilization and aptamer isolation, the proposed SELEX method allowed for efficient selection of glycoprotein-binding aptamers by 6 rounds and the dissociation constants were at 10(-8) M level. Because of the employment of boronate affinity monolithic capillary, the new SELEX approach overcame the above-mentioned drawbacks and provided several significant advantages, including rapid selection speed (only 2 days were needed), high specificity toward the target molecules, and minute reagent consumption.

[1]  M. Radrizzani,et al.  Specific oligobodies against ERK-2 that recognize both the native and the denatured state of the protein. , 2001, Journal of immunological methods.

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

[3]  J. Szostak,et al.  Selection in vitro of single-stranded DNA molecules that fold into specific ligand-binding structures , 1992, Nature.

[4]  B. Han,et al.  High performance aptamer affinity chromatography for single-step selective extraction and screening of basic protein lysozyme. , 2012, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[5]  J. Szostak,et al.  In vitro selection of RNA molecules that bind specific ligands , 1990, Nature.

[6]  J. Fréchet,et al.  RIGID MACROPOROUS POLYMER MONOLITHS , 1999 .

[7]  Michael Musheev,et al.  Nonequilibrium capillary electrophoresis of equilibrium mixtures: a universal tool for development of aptamers. , 2005, Journal of the American Chemical Society.

[8]  M. Breadmore,et al.  Boronate functionalised polymer monoliths for microscale affinity chromatography. , 2006, The Analyst.

[9]  Michael Musheev,et al.  Non-SELEX selection of aptamers. , 2006, Journal of the American Chemical Society.

[10]  M. Bowser,et al.  In vitro selection of high-affinity DNA ligands for human IgE using capillary electrophoresis. , 2004, Analytical chemistry.

[11]  Yuqi Feng,et al.  Boronate affinity monolith for highly selective enrichment of glycopeptides and glycoproteins. , 2009, The Analyst.

[12]  Sergey N Krylov,et al.  Non-SELEX: selection of aptamers without intermediate amplification of candidate oligonucleotides , 2006, Nature Protocols.

[13]  Gwo-Bin Lee,et al.  An integrated microfluidic system for rapid screening of alpha-fetoprotein-specific aptamers. , 2012, Biosensors & bioelectronics.

[14]  Hongyuan Chen,et al.  Ring-opening polymerization with synergistic co-monomers: access to a boronate-functionalized polymeric monolith for the specific capture of cis-diol-containing biomolecules under neutral conditions. , 2009, Angewandte Chemie.

[15]  A. Pardi,et al.  High-resolution molecular discrimination by RNA. , 1994, Science.

[16]  T. Ikegami,et al.  Monolithic columns for high-efficiency HPLC separations. , 2004, Current opinion in chemical biology.

[17]  Yunchun Liu,et al.  Synthesis of hydrophilic boronate affinity monolithic capillary for specific capture of glycoproteins by capillary liquid chromatography. , 2009, Journal of chromatography. A.

[18]  X Chris Le,et al.  Aptamer-modified monolithic capillary chromatography for protein separation and detection. , 2008, Analytical chemistry.

[19]  T. Block,et al.  Fucosylated Glycoproteins as Markers of Liver Disease , 2009, Disease markers.

[20]  X Chris Le,et al.  Aptamer-based affinity chromatographic assays for thrombin. , 2008, Analytical chemistry.

[21]  J. Weinstein,et al.  Biomarkers in Cancer Staging, Prognosis and Treatment Selection , 2005, Nature Reviews Cancer.

[22]  Jijun Tang,et al.  The DNA aptamers that specifically recognize ricin toxin are selected by two in vitro selection methods , 2006, Electrophoresis.

[23]  Yunchun Liu,et al.  A benzoboroxole-functionalized monolithic column for the selective enrichment and separation of cis-diol containing biomolecules. , 2012, Chemical communications.

[24]  M. Bowser,et al.  In vitro selection of aptamers with affinity for neuropeptide Y using capillary electrophoresis. , 2005, Journal of the American Chemical Society.

[25]  H. Zou,et al.  Synthesis and characterization of a new boronate affinity monolithic capillary for specific capture of cis-diol-containing compounds. , 2009, Journal of chromatography. A.

[26]  S. Gopinath Methods developed for SELEX , 2006, Analytical and bioanalytical chemistry.

[27]  H. Zou,et al.  Octadecylated Silica Monolith Capillary Column with Integrated Nanoelectrospray Ionization Emitter for Highly Efficient Proteome Analysis* , 2006, Molecular & Cellular Proteomics.

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

[29]  Mary Ann Comunale,et al.  GP73, a resident Golgi glycoprotein, is a novel serum marker for hepatocellular carcinoma. , 2005, Journal of hepatology.

[30]  M. Bowser,et al.  Capillary electrophoresis-SELEX selection of catalytic DNA aptamers for a small-molecule porphyrin target. , 2013, Analytical chemistry.

[31]  Yunchun Liu,et al.  A unique boronic acid functionalized monolithic capillary for specific capture, separation and immobilization of cis-diol biomolecules. , 2011, Chemical communications.

[32]  Michel H M Eppink,et al.  Alternative affinity tools: more attractive than antibodies? , 2011, The Biochemical journal.

[33]  M. Yarus,et al.  Selection of an RNA domain that binds Zn2+. , 1995, RNA.

[34]  Zhen Liu,et al.  Restricted access boronate affinity porous monolith as a protein A mimetic for the specific capture of immunoglobulin G , 2012 .

[35]  H. Zou,et al.  Coupling strong anion-exchange monolithic capillary with MALDI-TOF MS for sensitive detection of phosphopeptides in protein digest. , 2010, Analytical chemistry.

[36]  Jean M. J. Fréchet,et al.  New Designs of Macroporous Polymers and Supports: From Separation to Biocatalysis , 1996, Science.