Detection of Single Oligonucleotide by an Aerolysin Nanopore

Cao, Chunhua; Liao, Dong-Fang; Ying, Yi‐Lun; Long, Yi‐Tao

doi:10.6023/a16070352

Cited by 13 publications

(10 citation statements)

References 15 publications

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“…Next, the voltage‐dependent studies were conducted which revealed that the duration of Teln decreased exponentially with the applied voltage ranging from +120 to +180 mV ( Figure a). These results were consistent with our previous researches and confirmed the translocation of Teln through the aerolysin nanopore. The free energy barrier is associated with the transport of oligonucleotides.…”

Section: Statistical Results Of the Duration And The I/i0 For Teln (Nsupporting

confidence: 94%

“…Next, the voltage-dependent studies were conducted which revealed that the duration of Teln decreased exponentially with the applied voltage ranging from +120 to +180 mV (Figure 3a). These results were consistent with our previous researches [8,[26][27][28] Small 2018, 14, 1704520 Figure 2. Translocation of unfolded Teln (n = 1, 2, 3, 4, and 5) through an aerolysin nanopore in MgCl 2 solution.…”

Section: Doi: 101002/smll201704520supporting

confidence: 93%

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A General Strategy of Aerolysin Nanopore Detection for Oligonucleotides with the Secondary Structure

Liao

Cao

Ying

et al. 2018

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An aerolysin nanopore is employed as a sensitive tool for single-molecule analysis of short oligonucleotides (≤10 nucleotides), poly(ethylene glycol) (PEGs), peptides, and proteins. However, the direct analysis of long oligonucleotides with the secondary structure (e.g., G-quadruplex topology) remains a challenge, which impedes the further practical applications of the aerolysin nanopore. Here, a simple and applicable method of aerolysin nanopore is presented to achieve a direct analysis of structured oligonucleotides that are extended to 30 nucleotides long by a cation-regulation mechanism. By regulating the cation type in electrolyte solution, the structured oligonucleotides are unfolded into linear form which ensures the successive translocation. The results show that each model oligonucleotide of 5'-(TTAGGG) -3' can produce a well-resolved current blockade in its unfolded solution of MgCl . The length between 6 and 30 nucleotides long of model oligonucleotides is proportional to the duration time, showing a translocation velocity as low as 0.70-0.13 ms nt at +140 mV. This method exhibits an excellent sensitivity and a sufficient temporal resolution, provides insight into the aerolysin nanopore methodology for genetic and epigenetic biosensing, making aerolysin applicable in practical diagnosing with long and structured nucleic acids.

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Section: Statistical Results Of the Duration And The I/i0 For Teln (Nsupporting

confidence: 94%

Section: Doi: 101002/smll201704520supporting

confidence: 93%

A General Strategy of Aerolysin Nanopore Detection for Oligonucleotides with the Secondary Structure

Liao

Cao

Ying

et al. 2018

Small

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“…In 1996, Kasianowicz and his colleagues reported on the application of a-hemolysin nanochannels in nucleic acid detection for the first time at NIST [3]. So far, researchers have also developed various new biological nanochannels [4][5][6][7]. Nanochannels single-channel detection technology has also been applied to DNA sequencing [4,6], peptide detection [8][9][10] and RNA detection [11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

“…So far, researchers have also developed various new biological nanochannels [4][5][6][7]. Nanochannels single-channel detection technology has also been applied to DNA sequencing [4,6], peptide detection [8][9][10] and RNA detection [11][12][13][14]. Due to the physical occupancy effect of biomolecules, when the biomolecules pass through the nanochannel, the blockage will cause a square wave signal, and when the molecular length is long, the modulation current caused by the lengthening will become longer.…”

Section: Introductionmentioning

confidence: 99%

A new method for early detection of pancreatic cancer biomarkers: detection of microRNAs by nanochannels

Liao

et al. 2019

Artificial Cells, Nanomedicine, and Biotechnology

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Objective: To develop an effective new method for early detection of pancreatic cancer biomarkers and to aid early clinical diagnosis. Methods: A DNA probe (Probe) capable of specifically recognizing the target miRNA was designed. The specific probe of miRNA 21 is designed first, and then mixed with the miRNA 21 sample to form a complex molecule, and the complex molecule is added to the nanochannels to detect the received signal. The probe is designed to detect the electrical signal by means of pre-matching and post-matching and observe the stability of the signal. The miRNA 21, miRNA 155, miRNA 196a were added to the nano-single channel to detect the characteristic signals and blocking time. The miRNA 21Áprobe 21 mixture was mixed with other five cancer-associated microRNAs, and the signal results of the detection were collected and compared. Results: The signal of miRNA 21 was successfully detected. Whether the probe is designed at the front or the back, there are two signal results. The Probe should be designed to match the middle region of the miRNA. The three microRNA complex molecules have different characteristic signals and blocking times, which can be effectively distinguished. Conclusion: Nanochannels can effectively detect pancreatic cancer-related microRNAs.

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“…针对分子个体的检测技术如原子力显微镜(AFM)受限于样品状态和荧光标记, 无法检测液相样品. 而与传统蛋白质检测手段相比, 新兴的纳米孔技术 [16,17] 的优势在于: (1)无标志检测; (2)针对分子个体而不是蛋白分子全体.…”

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Label-free Detection of PD-1 Antibody and Antigen Immunoreaction Using Nano-Sensors

Fu¹,

Zhang²,

Sun³

et al. 2019

Acta Chim. Sinica

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Immunotherapy for cancer is a method to treat cancer by using the body's own immune system. Programmed death receptor 1 (PD-1) is one of the checkpoints in the immunotherapy. The signal pathway PD-1 (programmed death receptor 1)/PD-L1 (ligand of PD-1) is closely related to the immune escape of the cancer cells. The inhibitor drugs for PD-1 checkpoint, essentially the monoclonal antibodies of PD-1 or PD-L1 which is essentially the immune checkpoints inhibitors could block the PD-1/PD-L1 pathway and reactivate T-cells to kill cancer cells, and as a result, the immunotherapy for cancer is realized. In order to study the binding process of PD-1 drugs and PD-1 antigen in vivo, in this work, solid-state nanopore as a single molecule method is used to detect the binding of PD-1 antibody and antigen. The PD-1 antibody as well as antigen is driven though the same nanopore under the same experimental condition by the external electric field. Since the antibody's block is about 0.01297 while the antigen's block is 0.00404, the PD-1 antibody is distinguished with the PD-1 antigen according to the theoretical formula. Driving the PD-1 antigen though the nanopore modified by PD-1 antibody (a series of experiments are conducted for characterization) under the same temperature and buffer concentration, the antibody-antigen complexes are detected and distinguished with PD-1 protein and its antibody through the relative current drop analysis and the current drop achieved before. The results suggest that the antibody and antigen have a specific binding (the smaller peak represents the free PD-1 antibody and antigen) and the binding process can be detected by nano-sensors. So the nanopore is able to distinguish the antibody, the antigen and the complexes without any labling. And in the future, the nanopore technology may be a rapid and label-free way for patients and doctors to evaluate the drugs' efficiency.

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Detection of Single Oligonucleotide by an Aerolysin Nanopore

Cited by 13 publications

References 15 publications

A General Strategy of Aerolysin Nanopore Detection for Oligonucleotides with the Secondary Structure

A General Strategy of Aerolysin Nanopore Detection for Oligonucleotides with the Secondary Structure

A new method for early detection of pancreatic cancer biomarkers: detection of microRNAs by nanochannels

Label-free Detection of PD-1 Antibody and Antigen Immunoreaction Using Nano-Sensors

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