Cell-SELEX Technology

Ohuchi, Shoji

doi:10.1089/biores.2012.0253

Cited by 121 publications

(92 citation statements)

References 52 publications

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“…5,6,7,8 Aside from their high affinities, the exquisite specificities of aptamers suggested binding of precisely folded entities that recognize their targets through shape complementarity. This notion is now corroborated with several crystal structures of aptamer–target complexes.…”

Section: Introductionmentioning

confidence: 99%

Nucleic Acid Ligands With Protein-like Side Chains: Modified Aptamers and Their Use as Diagnostic and Therapeutic Agents

Rohloff

Gelinas

Jarvis

et al. 2014

Molecular Therapy - Nucleic Acids

445

391

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Limited chemical diversity of nucleic acid libraries has long been suspected to be a major constraining factor in the overall success of SELEX (Systematic Evolution of Ligands by EXponential enrichment). Despite this constraint, SELEX has enjoyed considerable success over the past quarter of a century as a result of the enormous size of starting libraries and conformational richness of nucleic acids. With judicious introduction of functional groups absent in natural nucleic acids, the “diversity gap” between nucleic acid–based ligands and protein-based ligands can be substantially bridged, to generate a new class of ligands that represent the best of both worlds. We have explored the effect of various functional groups at the 5-position of uracil and found that hydrophobic aromatic side chains have the most profound influence on the success rate of SELEX and allow the identification of ligands with very low dissociation rate constants (named Slow Off-rate Modified Aptamers or SOMAmers). Such modified nucleotides create unique intramolecular motifs and make direct contacts with proteins. Importantly, SOMAmers engage their protein targets with surfaces that have significantly more hydrophobic character compared with conventional aptamers, thereby increasing the range of epitopes that are available for binding. These improvements have enabled us to build a collection of SOMAmers to over 3,000 human proteins encompassing major families such as growth factors, cytokines, enzymes, hormones, and receptors, with additional SOMAmers aimed at pathogen and rodent proteins. Such a large and growing collection of exquisite affinity reagents expands the scope of possible applications in diagnostics and therapeutics.

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Section: Introductionmentioning

confidence: 99%

Nucleic Acid Ligands With Protein-like Side Chains: Modified Aptamers and Their Use as Diagnostic and Therapeutic Agents

Rohloff

Gelinas

Jarvis

et al. 2014

Molecular Therapy - Nucleic Acids

445

391

View full text Add to dashboard Cite

show abstract

“…25,26 More recently, nucleic acid aptamers have been introduced as novel recognition elements. [27][28][29][30][31] Aptamers are short artificial DNA and RNA sequences selectively chosen by their extremely strong binding affinity to their target proteins through a process called Systematic Evolution of Ligands by Exponential Enrichment (SELEX). In the SELEX process, an extremely large pool of oligonucleotides of different sequences is incubated with the protein target of interest.…”

Section: Preliminary Results and Discussionmentioning

confidence: 99%

Functionalized paper SERS (P-SERS) substrates for selective targeting of analytes in complex samples

Wang¹,

Hoppmann²

2015

SPIE Proceedings

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Surface enhanced Raman spectroscopy (SERS) requires the analyte molecule to be close to the plasmonic surface in order to generate SERS enhancement. This limitation restricts the practical application of SERS to molecules that possess functional groups that interact strongly with gold or silver surfaces. Moreover, the identification of target analytes in a complex sample matrix is made even more difficult when interferents compete with the target for binding to the plasmonic surface, resulting in overlapping spectral signatures. In this work, we report a strategy to functionalize inkjet printed P-SERS substrates by strategically placing supramolecular structures (such as nucleic acid aptamers) onto the gold nanoparticles. This promotes the selective interaction of target molecules with the plasmonic surface, leading to improved sensor performance.

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“…For the improvement of specificity to targets, counter-SELEX has been developed, in which only unbounded nucleotides to non-target bacteria were selected and amplified for eliminating undesired aptamers. However, complete elimination of variants binding to nonspecific molecules are difficult even though repeated process of SELEX and counter-SELEX (Ohuchi, 2012). Therefore, the post-SELEX modifications are suggested in order to enhance the selectivity and binding affinity.…”

Section: Whole-cell Selexmentioning

confidence: 99%

The Importance of FACS Analysis in the Development of Aptamers Specific to Pathogens

Moon¹,

Kim²,

Park³

et al. 2014

Journal of Biosystems Engineering

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Purpose: This review aims to introduce aptamers and the methods of its development to improve the sensitivity and selectivity to target bacteria. In this review, we have highlighted current developments and directions in the pathogen detection based on aptamers. Background: Aptamers, the specific nucleic acid sequences, can bind to targets with high affinity and specificity. Some of researches on the use of aptamers for the detection of pathogen have been reported in recent years. Aptamers have more applicability than antibodies for the development of pathogen detection using biosensor; such as easy to synthesis and labeling, lack of immunogenicity, and a low cost of production. However, only few reports on the development and use of aptamers for the detection of pathogen have been published. Review: Aptamers specific to pathogen are obtained by whole-cell systematic evolution of ligands by exponential enrichment (SELEX) process. SELEX process is composed of screening random oligonucleotide bound with target cells, multiple separation and amplification of nucleic acids, final identification of the best sequences. For improving those affinity and selectivity to target bacteria, optimization of multiple separating process to remove unbounded oligonucleotides from aptamer candidates and sorting process by flow cytometry are required.

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Cell-SELEX Technology

Cited by 121 publications

References 52 publications

Nucleic Acid Ligands With Protein-like Side Chains: Modified Aptamers and Their Use as Diagnostic and Therapeutic Agents

Nucleic Acid Ligands With Protein-like Side Chains: Modified Aptamers and Their Use as Diagnostic and Therapeutic Agents

Functionalized paper SERS (P-SERS) substrates for selective targeting of analytes in complex samples

The Importance of FACS Analysis in the Development of Aptamers Specific to Pathogens

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