Isolation and Characterization of High Affinity Aptamers Against DNA Polymerase Iota

Лахин, А. В.; Kazakov, Andrei A.; Makarova, Аlena V.; Pavlov, Youri I.; Ефремова, А.; Shram, S. I.; Тарантул, В. З.; Генинг, Л. В.

doi:10.1089/nat.2011.0324

Cited by 15 publications

(5 citation statements)

References 29 publications

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“…Due to their relatively accessible synthetic process, flexibility in design, and excellent molecular interaction profile, they have found use in biotechnological and therapeutic applications. [8,9] They offer advantages over antibodies because they are readily produced by chemical synthesis; possess desirable storage properties and elicit little or no immunogenicity in therapeutic applications. [6] Despite this, they do have some limitations, nominally around their thermal and chemical stability-the nature of the nucleic acid sequence leaves it open to degradation.…”

Section: Introductionmentioning

confidence: 99%

Hybrid Aptamer‐Molecularly Imprinted Polymer (aptaMIP) Nanoparticles from Protein Recognition—A Trypsin Model

Sullivan

Clay

Moazami

et al. 2021

Macromolecular Bioscience

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Aptamers offer excellent potential for replacing antibodies for molecular recognition purposes however their performance can compromise with biological/environmental degradation being a particular problem. Molecularly imprinted Polymers (MIPs) offer an alternative to biological materials and while these offer the robustness and ability to work in extreme environmental conditions, they often lack the same recognition performance. By slightly adapting the chemical structure of a DNA aptamer it is incorporated for use as the recognition part of a MIP, thus creating an aptamer‐MIP hybrid or aptaMIP. Here these are developed for the detection of the target protein trypsin. The aptaMIP nanoparticles offer superior binding affinity over conventional MIP nanoparticles (nanoMIPs), with KD values of 6.8 × 10−9 (±0.2 × 10−9) m and 12.3 × 10−9 (±0.4 × 10−9) m for the aptaMIP and nanoMIP, respectively. The aptaMIP also outperforms the aptamer only (10.3 × 10−9 m). Good selectivity against other protein targets is observed. Using surface plasmon resonance, the limit of detection for aptaMIP nanoparticles is twofold lower (2 nm) compared to the nanoMIP (4 nm). Introduction of the aptamer as a “macro‐monomer” into the MIP scaffold has beneficial effects and offers potential to improve this class of polymers significantly.

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Section: Introductionmentioning

confidence: 99%

Hybrid Aptamer‐Molecularly Imprinted Polymer (aptaMIP) Nanoparticles from Protein Recognition—A Trypsin Model

Sullivan

Clay

Moazami

et al. 2021

Macromolecular Bioscience

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“…The Pol ι enzymatic activity was estimated by the appearance of an additional specific band on electropherograms, which corresponds to the 18-mer product with misincorporated G opposite template T. Only Pol ι is capable of generating significant amounts of this product, since its formation is inhibited by Pol ι-specific aptamer [ 26 ]. Relying on this unique feature we have developed a method for the detection of Pol ι activity in cell extracts, named misGvA (misincorporation of G versus A) [ 24 , 25 , 27 , 28 ].…”

Section: Resultsmentioning

confidence: 99%

Alterations in Synthesis and Repair of DNA during the Development of Loach Misgurnus fossilis

Генинг

Лахин

Makarova

et al. 2016

JDB

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Using a modified radiolabeled primer extension method (we named this modification misGvA—“misincorporation of G versus A”) we have investigated the DNA synthesis and repair at early and late stages of development of loach Misgurnus fossilis. The misincorporation activity of DNA polymerase iota (Pol ι) in wild-type loach could not be detected by this method at any stage of loach development. In transgenic loach overexpressing human Pol ι we have shown that the bypassing of DNA synthesis arrest after incorporation of mismatched nucleotide by Pol ι (the T-stop) was not associated with this enzyme. Non-transgenic loach larvae are virtually lacking the capacity for error correction of DNA duplex containing a mismatched nucleotide. Such repair activity develops only in the adult fish. It appears that the initial stages of development are characterized by more intensive DNA synthesis, while in terminal stages the repair activities become more prominent. The misGvA approach clearly indicates substantial changes in the DNA synthesis intensity, although the role of particular replicative and repair DNA polymerases in this process requires further study.

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“…The alternative, using the already existing functional groups further decreases the number of functional groups available for aptamer target recognition. Counter-selection steps should be carried out in SELEX to increase specificity and exclude cross-reactivity [ 79 , 80 ]. However, in an already time-consuming protocol these extra steps may drastically worsen the economics.…”

Section: Challenges In the Detection Of Small Molecules Using Aptamentioning

confidence: 99%

Aptasensors for Point-of-Care Detection of Small Molecules

et al. 2020

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Aptamers, a group of nucleic acids which can specifically bind to a target molecule, have drawn extensive interest over the past few decades. For analytics, aptamers represent a viable alternative to gold-standard antibodies due to their oligonucleic nature combined with advantageous properties, including higher stability in harsh environments and longer shelf-life. Indeed, over the last decade, aptamers have been used in numerous bioanalytical assays and in various point-of-care testing (POCT) platforms. The latter allows for rapid on-site testing and can be performed outside a laboratory by unskilled labor. Aptamer technology for POCT is not limited just to medical diagnostics; it can be used for a range of applications, including environmental monitoring and quality control. In this review, we critically examine the use of aptamers in POCT with an emphasis on their advantages and limitations. We also examine the recent success of aptasensor technology and how these findings pave the way for the analysis of small molecules in POCT and other health-related applications. Finally, the current major limitations of aptamers are discussed, and possible approaches for overcoming these challenges are presented.

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Isolation and Characterization of High Affinity Aptamers Against DNA Polymerase Iota

Cited by 15 publications

References 29 publications

Hybrid Aptamer‐Molecularly Imprinted Polymer (aptaMIP) Nanoparticles from Protein Recognition—A Trypsin Model

Hybrid Aptamer‐Molecularly Imprinted Polymer (aptaMIP) Nanoparticles from Protein Recognition—A Trypsin Model

Alterations in Synthesis and Repair of DNA during the Development of Loach Misgurnus fossilis

Aptasensors for Point-of-Care Detection of Small Molecules

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