PEP-on-DEP: A competitive peptide-based disposable electrochemical aptasensor for renin diagnostics

Biyani, Manish; Kawai, Keiko; Kitamura, Koichiro; Chikae, Miyuki; Biyani, Madhu; Ushijima, Hiromi; Tamiya, Eiichi; Yoneda, Takashi; Takamura, Yuzuru

doi:10.1016/j.bios.2015.12.078

Cited by 22 publications

(10 citation statements)

References 26 publications

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“…Other potential applications of this methodology include the development of biosensors for previously unreachable biomarkers or the intelligent design of peptide aptamers. Certain small protein targets such as the medically relevant angiotensinogenase renin have proven difficult to probe with traditional approaches but can be detected using peptide-based aptamers developed using cDNA display techniques (Biyani et al., 2016). Peptides designed to function as therapeutics can outperform relatively larger (∼150 kDa) antibodies in terms of bioavailability, tumor penetration, and production efficiency and have been used to develop binding assays, cancer therapy, drug delivery, and in vivo imaging (Yu et al., 2017).…”

Section: Discussionmentioning

confidence: 99%

In Silico Engineering of Synthetic Binding Proteins from Random Amino Acid Sequences

et al. 2019

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SummarySynthetic proteins with high affinity and selectivity for a protein target can be used as research tools, biomarkers, and pharmacological agents, but few methods exist to design such proteins de novo. To this end, the In-Silico Protein Synthesizer (InSiPS) was developed to design synthetic binding proteins (SBPs) that bind pre-determined targets while minimizing off-target interactions. InSiPS is a genetic algorithm that refines a pool of random sequences over hundreds of generations of mutation and selection to produce SBPs with pre-specified binding characteristics. As a proof of concept, we design SBPs against three yeast proteins and demonstrate binding and functional inhibition of two of three targets in vivo. Peptide SPOT arrays confirm binding sites, and a permutation array demonstrates target specificity. Our foundational approach will support the field of de novo design of small binding polypeptide motifs and has robust applicability while offering potential advantages over the limited number of techniques currently available.

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Section: Discussionmentioning

confidence: 99%

In Silico Engineering of Synthetic Binding Proteins from Random Amino Acid Sequences

et al. 2019

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“…Many electrochemical techniques are used in analytical chemistry. The most commonly used ones for microfluidic devices or aptamer biosensors are amperometry [ 43 ], voltammetry [ 31 , 34 , 40 , 44 , 80 , 85 ], and electrochemical impedance spectroscopy [ 32 , 35 , 72 , 86 , 87 , 88 , 89 ]. Liu et al [ 87 ] developed ZnO/graphene (ZnO/G) composite with S6 aptamer for a photoelectrochemical (PEC) detector.…”

Section: Detection Methods and Assay Formatsmentioning

confidence: 99%

Recent Microdevice-Based Aptamer Sensors

et al. 2018

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Since the systematic evolution of ligands by exponential enrichment (SELEX) method was developed, aptamers have made significant contributions as bio-recognition sensors. Microdevice systems allow for low reagent consumption, high-throughput of samples, and disposability. Due to these advantages, there has been an increasing demand to develop microfluidic-based aptasensors for analytical technique applications. This review introduces the principal concepts of aptasensors and then presents some advanced applications of microdevice-based aptasensors on several platforms. Highly sensitive detection techniques, such as electrochemical and optical detection, have been integrated into lab-on-a-chip devices and researchers have moved towards the goal of establishing point-of-care diagnoses for target analyses.

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“…In this paper, we showed the plausibility of using SELCOS on close targets of influenza virus subtypes (H1N1 and H3N2). We also introduced a DEPSOR-mode electrochemical sensing method (or Apta-DEPSOR) 18,19 for readily evaluating the specific binding of aptamers. On the whole, a powerful approach for rapidly detecting various influenza subtypes with high sensitivity is presented here, and it addresses several theoretical considerations.…”

Section: Introductionmentioning

confidence: 99%

Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor

Kushwaha

Takamura

Nishigaki

et al. 2019

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The SELEX (Systematic Evolution of Ligands by EXponential enrichment) method has been used successfully since 1990, but work is still required to obtain highly specific aptamers. Here, we present a novel approach called ‘Competitive non-SELEX’ (and termed as ‘SELCOS’ (Systematic Evolution of Ligands by COmpetitive Selection)) for readily obtaining aptamers that can discriminate between highly similar targets. This approach is based on the theoretical background presented here, in which under the co-presence of two similar targets, a specific binding type can be enriched more than a nonspecifically binding one during repetitive steps of partitioning with no PCR amplification between them. This principle was experimentally confirmed by the selection experiment for influenza virus subtype-specific DNA aptamers. Namely, the selection products (pools of DNA aptamers) obtained by SELCOS were subjected to a DEPSOR-mode electrochemical sensor, enabling the method to select subtype-specific aptamer pools. From the clonal analysis of these pools, only a few rounds of in vitro selection were sufficient to achieve the surprisingly rapid enrichment of a small number of aptamers with high selectivity, which could be attributed to the SELCOS principle and the given selection pressure program. The subtype-specific aptamers obtained in this manner had a high affinity (e.g., K D = 82 pM for H1N1; 88 pM for H3N2) and negligible cross-reactivity. By making the H1N1-specific DNA aptamer a sensor unit of the DEPSOR electrochemical detector, an influenza virus subtype-specific and portable detector was readily constructed, indicating how close it is to the field application goal.

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PEP-on-DEP: A competitive peptide-based disposable electrochemical aptasensor for renin diagnostics

Cited by 22 publications

References 26 publications

In Silico Engineering of Synthetic Binding Proteins from Random Amino Acid Sequences

In Silico Engineering of Synthetic Binding Proteins from Random Amino Acid Sequences

Recent Microdevice-Based Aptamer Sensors

Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor

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