Design of Peptide Ligands (Aptamers) for Determination of Myeloperoxidase Level in Blood Using Biochips

Sitkov, Nikita O.; Zimina, Tatiana M.; Карасев, В. Е.; Lemozerskii, Vladislav E.; Kolobov, A. A.

doi:10.1109/eiconrus49466.2020.9039329

Cited by 7 publications

(5 citation statements)

References 16 publications

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“…The development of peptide aptamer sequences could be carried out using various in-silico modeling methods [1]. In this work, the modeling was carried out using the "Protein 3D" visualizer of supramolecular structures [2], developed at the Center of Microtechnology and Diagnostics of St. Petersburg State Electrotechnical University.…”

Section: Methodsmentioning

confidence: 99%

Label-Free Peptide-Based Biosensor for Express Detection of Protein Markers of Acute Cardiovascular Conditions in Biological Fluids

Sitkov

Zimina

Карасев

et al. 2020

The 1st International Electronic Conference on Biosensors

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Acute cardiovascular conditions require prompt assistance, which depends on a timely and accurate diagnosis. This could be achieved by using biosensor systems based on peptide aptamers capable of selectively binding protein markers of diseases. In this work, a label-free biosensor system based on fluorometric registration of the formation of a “peptide aptamer—target protein” complex is considered. It comprises a microfluidic subsystem integrated with arrays of sites with immobilized peptide aptamers, coupled with an optical detection system. The clinical sample of the whole blood is loaded into the inlet basin, where the cells are separated and plasma flows into the microfluidic channel for analysis. Peptide aptamers were created using the molecular complement search technique based on the search for systems of conjugated ion-hydrogen bonds in the three-dimensional structures of target proteins. The technology for manufacturing a microfluidic chip is a combination of thick-film and photolithography technologies based on the SU-8 photoresist, for which the relief and surface morphology have been studied. The composition of the biochip layers is selected in such a way that ultraviolet light with a wavelength of 280 nm passes through an inlet window, excites fluorescence inside the channel, which passes through the glass window, which absorbs UV-light. This wavelength accounts for the maximum absorption of aromatic amino acids—tyrosine and tryptophan. In this case, one of the last layers is a luminophore layer for re-emission of protein fluorescence as a visible light. The reading platform includes a 280 nm LED, a video sensor, 3D-printed PLA tooling, and software for processing and analyzing the received signal.

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Section: Methodsmentioning

confidence: 99%

Label-Free Peptide-Based Biosensor for Express Detection of Protein Markers of Acute Cardiovascular Conditions in Biological Fluids

Sitkov

Zimina

Карасев

et al. 2020

The 1st International Electronic Conference on Biosensors

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“…The peptide aptamers were designed using data from Protein Data Bank [40] and "Protein 3D" software, developed at the Center of Microtechnology and Diagnostics of St. Petersburg Electrotechnical University «LETI» [41][42][43]. Two versions of peptides designed for troponin T were considered.…”

Section: Peptide Aptamersmentioning

confidence: 99%

“…The representation of proteins and peptides in the form of CIHBS has proved to be useful as a technology for in silico development of peptide aptamers. Based on this technology, a number of peptide sequences of 15-30 amino acid residues have been proposed and synthesized, and their ability of selective binding target proteins was experimentally tested using capillary electrophoresis [43,50].…”

Section: Designing Peptide Aptamers For Label-free Detectionmentioning

confidence: 99%

Toward Development of a Label-Free Detection Technique for Microfluidic Fluorometric Peptide-Based Biosensor Systems

et al. 2021

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The problems of chronic or noncommunicable diseases (NCD) that now kill around 40 million people each year require multiparametric combinatorial diagnostics for the selection of effective treatment tactics. This could be implemented using the biosensor principle based on peptide aptamers for spatial recognition of corresponding protein markers of diseases in biological fluids. In this paper, a low-cost label-free principle of biomarker detection using a biosensor system based on fluorometric registration of the target proteins bound to peptide aptamers was investigated. The main detection principle considered includes the re-emission of the natural fluorescence of selectively bound protein markers into a longer-wavelength radiation easily detectable by common charge-coupled devices (CCD) using a specific luminophore. Implementation of this type of detection system demands the reduction of all types of stray light and background fluorescence of construction materials and aptamers. The latter was achieved by careful selection of materials and design of peptide aptamers with substituted aromatic amino acid residues and considering troponin T, troponin I, and bovine serum albumin as an example. The peptide aptamers for troponin T were designed in silico using the «Protein 3D» (SPB ETU, St. Petersburg, Russia) software. The luminophore was selected from the line of ZnS-based solid-state compounds. The test microfluidic system was arranged as a flow through a massive of four working chambers for immobilization of peptide aptamers, coupled with the optical detection system, based on thick film technology. The planar optical setup of the biosensor registration system was arranged as an excitation-emission cascade including 280 nm ultraviolet (UV) light-emitting diode (LED), polypropylene (PP) UV transparent film, proteins layer, glass filter, luminophore layer, and CCD sensor. A laboratory sample has been created.

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“…Of considerable interest is the research and development of the technological foundations of the heterogeneous integration of an artificial biorecognition element that performs the function of spatial recognition of the target structure [8] into the hybrid structure of the biosensor system and ensures a unified integral-group technological cycle of its formation. The atomic-molecular design of a biorecognition element allows the editing of its physical properties in order to improve the functional characteristics of the detection system [23][24][25]. Peptide aptamers (oligopeptides built of 8-40 amino acid residues) can serve as biorecognition elements with a high degree of affinity and a specificity of complex formation with target protein macromolecules.…”

Section: Introductionmentioning

confidence: 99%

“…The complexes of peptide aptamers and target proteins have dissociation constants comparable, and are sometimes better than those of antigen-antibody complexes [23]. Therefore, they are extremely promising for the development of biosensors that aim to detect the protein markers of diseases [24][25][26][27].…”

Section: Introductionmentioning

confidence: 99%

Study of the Fabrication Technology of Hybrid Microfluidic Biochips for Label-Free Detection of Proteins

et al. 2021

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A study of the peculiarities and a comparative analysis of the technologies used for the fabrication of elements of novel hybrid microfluidic biochips for express biomedical analysis have been carried out. The biochips were designed with an incorporated microfluidic system, which enabled an accumulation of the target compounds in a biological fluid to be achieved, thus increasing the biochip system’s sensitivity and even implementing a label-free design of the detection unit. The multilevel process of manufacturing a microfluidic system of a given topology for label-free fluorometric detection of protein structures is presented. The technological process included the chemical modification of the working surface of glass substrates by silanization using (3-aminopropyl) trimethoxysilane (APTMS), formation of the microchannels, for which SU-8 technologies and a last generation dry film photoresist were studied and compared. The solid-state phosphor layers were deposited using three methods: drop application; airbrushing; and mechanical spraying onto the adhesive surface. The processes of sealing the system, installing input ports, and packaging using micro-assembly technologies are described. The technological process has been optimized and the biochip was implemented and tested. The presented system can be used to design novel high-performance diagnostic tools that implement the function of express detection of protein markers of diseases and create low-power multimodal, highly intelligent portable analytical decision-making systems in medicine.

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Design of Peptide Ligands (Aptamers) for Determination of Myeloperoxidase Level in Blood Using Biochips

Cited by 7 publications

References 16 publications

Label-Free Peptide-Based Biosensor for Express Detection of Protein Markers of Acute Cardiovascular Conditions in Biological Fluids

Label-Free Peptide-Based Biosensor for Express Detection of Protein Markers of Acute Cardiovascular Conditions in Biological Fluids

Toward Development of a Label-Free Detection Technique for Microfluidic Fluorometric Peptide-Based Biosensor Systems

Study of the Fabrication Technology of Hybrid Microfluidic Biochips for Label-Free Detection of Proteins

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