Real-time label-free assessment of T7 DNA polymerase immobilization

Dronina, Julija; Plaušinaitis, Deivis; Bubniene, U.; Ramanavičius, Arūnas

doi:10.1016/j.mtnano.2022.100232

Cited by 4 publications

(3 citation statements)

References 49 publications

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“…The QCM biosensor is an extremely sensitive mass sensor capable of measuring subnanogram levels of mass changes [ 144 , 145 , 146 , 147 ]. QCM biosensors are suitable for direct and label-free monitoring of affinity interactions of biomolecules [ 148 , 149 , 150 , 151 ]. In the efforts to develop a new strategy for simple, selective, and sensitive detection of SNPs, different designs including TSDR and nanomaterials-based QCM detection platforms have been reported [ 152 , 153 , 154 , 155 ].…”

Section: Biosensors For Snp Detectionmentioning

confidence: 99%

Recent Progress in Single-Nucleotide Polymorphism Biosensors

Wu,

Kong,

Zhang

et al. 2023

Biosensors

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Single-nucleotide polymorphisms (SNPs), the most common form of genetic variation in the human genome, are the main cause of individual differences. Furthermore, such attractive genetic markers are emerging as important hallmarks in clinical diagnosis and treatment. A variety of destructive abnormalities, such as malignancy, cardiovascular disease, inherited metabolic disease, and autoimmune disease, are associated with single-nucleotide variants. Therefore, identification of SNPs is necessary for better understanding of the gene function and health of an individual. SNP detection with simple preparation and operational procedures, high affinity and specificity, and cost-effectiveness have been the key challenge for years. Although biosensing methods offer high specificity and sensitivity, as well, they suffer drawbacks, such as complicated designs, complicated optimization procedures, and the use of complicated chemistry designs and expensive reagents, as well as toxic chemical compounds, for signal detection and amplifications. This review aims to provide an overview on improvements for SNP biosensing based on fluorescent and electrochemical methods. Very recently, novel designs in each category have been presented in detail. Furthermore, detection limitations, advantages and disadvantages, and challenges have also been presented for each type.

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Section: Biosensors For Snp Detectionmentioning

confidence: 99%

Recent Progress in Single-Nucleotide Polymorphism Biosensors

Wu,

Kong,

Zhang

et al. 2023

Biosensors

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“…Nucleic acids (DNAs and RNAs) are essential molecules for life on the earth and serve as the primary materials for storing, transmitting, and retrieving genetic information in living organisms. Due to their unique features, such as sequence programmability of structures and properties and good biocompatibility, DNAs and RNAs have been broadly used to develop novel materials and methods for biotechnology and biomedicine, including numerous biosensing tools. , The enzymes that are responsible for the synthesis and modification of DNAs and RNAs, such as DNA polymerases and Cas9 protein, have also been widely employed in the construction of biosensing systems. , In recent years, various unnatural modifications have been introduced into different moieties of nucleic acids and greatly expanded their structures, properties, and functions, which led to their broader application in related fields. , While many artificial modifications on the sugar-phosphate scaffold can greatly increase the chemical and biological stability of the nucleic acids, the artificial modifications on the nucleobases have been extensively employed to attach novel functionalities to the nucleic acids …”

Section: Introductionmentioning

confidence: 99%

“…1,2 The enzymes that are responsible for the synthesis and modification of DNAs and RNAs, such as DNA polymerases and Cas9 protein, have also been widely employed in the construction of biosensing systems. 3,4 In recent years, various unnatural modifications have been introduced into different moieties of nucleic acids and greatly expanded their structures, properties, and functions, which led to their broader application in related fields. 5,6 While many artificial modifications on the sugarphosphate scaffold can greatly increase the chemical and biological stability of the nucleic acids, the artificial modifications on the nucleobases have been extensively employed to attach novel functionalities to the nucleic acids.…”

Section: Introductionmentioning

confidence: 99%

Bottlebrush DNA-Primed Rolling Circle Amplification for Sensitive Detection of Biomolecules

Chen,

Zhang,

Peng

et al. 2023

ACS Appl. Nano Mater.

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Development of nucleic acids containing unnatural moieties has expanded the scope of genetic materials, and modifications of nucleobases have been extensively explored and found broad applications. However, previous efforts have mainly focused on modifying nucleobases with small molecules, while nucleobase modification with macromolecules has barely been explored. In this work, we first demonstrated the construction of highly programmable bottlebrush-like DNA structures by enzymatic production of DNA containing nucleobases modified with coupling handles and chemical attachment of single-stranded DNAs onto these handles. Employing these bottlebrush DNAs (BDs), we then developed a method named bottlebrush DNA-primed rolling circle amplification (BDP-RCA), which could rapidly produce branched DNA products with ultrahigh molecular weights in a controllable manner. BDP-RCA generated up to approximately 20-fold more products than ordinary RCA within 10 min. Moreover, the BDP-RCA product demonstrated a netlike structure in aqueous solution and a structure consisted of uniformly sized nanoflowers with a diameter of approximately 0.7 μm when lyophilized. These observations suggested the great potential of BDP-RCA in the development of materials and methods for biosensing, which were demonstrated with several examples. First, ultrasensitive detection of human α-thrombin with a limit of detection (LOD) of 1.1 pM was achieved by using BDP-RCA to amplify the detection signal in an ELISA-like assay. Second, magnetic beads immobilizing the BDP-RCA product were successfully applied for the efficient capture, enrichment, and preservation of a protein or bacterial cells to be detected. The LOD for the detection of human α-thrombin with this method was 68.5 pM.

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