Selecting DNA aptamers for endotoxin separation

Zhu, FangFang; Yu, Yi; Chen, Jianshu; Mei, Jianfeng; Zhang, Yanlu; Shuqing, Chen

doi:10.1007/s10529-015-1839-8

Cited by 5 publications

(3 citation statements)

References 11 publications

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“…It allows them to bind with high specificity and affinity to their targets. This was previously shown where aptamers could penetrate and be retained by tumor cells longer than antibodies, both in vitro and in vivo (50). As a result, they could be employed as molecular imaging probes (44).…”

Section: Discussionmentioning

confidence: 83%

Synthesis of fluorescence labelled aptamers for use as low‑cost reagents in HIV/AIDS research and diagnostics

et al. 2021

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Aptamers are nucleic acids selected by systematic evolution of ligands by exponential enrichment. They have potential as alternatives to antibodies in medical research and diagnostics, with the advantages of being non-immunogenic and relatively inexpensive to produce. In the present study, gp120 aptamers conjugated with fluorescein isothiocyanate (FITC) were generated, which could interact with HIV-1 gp120. A previously isolated gp120 aptamer, CSIR 1.1, was conjugated with FITC by incubation with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and imidazole. The conjugation and binding to the glycoprotein were confirmed by flow cytometry. FITC conjugated aptamers showed an increase in fluorescence emission 24-fold higher than baseline, and this difference was statistically significant (P=0.0016). Compared with a commercially available biotinylated anti-gp120 antibody, detected using FITC conjugated streptavidin, the emission of fluorescence obtained from the FITC-conjugated aptamer was 8-fold higher, suggesting a stronger interaction with gp120. In addition, the FITC conjugated aptamer neutralized HIV-1 pseudoviruses with an average IC 50 of 21.3 nM, similar to the parent aptamer that had an IC 50 of 19.2 nM. However, the difference in inhibition between the two aptamers was not statistically significant (P= 0.784). These results indicate that the FITC-conjugated aptamer generated in the present study could potentially be used as a low-cost reagent in HIV/AIDS research and diagnostics.

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Section: Discussionmentioning

confidence: 83%

Synthesis of fluorescence labelled aptamers for use as low‑cost reagents in HIV/AIDS research and diagnostics

et al. 2021

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“…In our previous study, the aptamer EAQ3 was selected by SELEX [12]. In the present study, EAQ3, which has an equilibrium dissociation constant (K D ) of 22.9 nM and a high affinity for endotoxin, is detected by biolayer interferometry.…”

Section: Introductionmentioning

confidence: 97%

Construction and application of an electrochemical biosensor based on an endotoxin aptamer

Wang

Chen

et al. 2017

Biotech and App Biochem

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An electrochemical biosensor that used an aptamer as a biological element was constructed to detect endotoxin. Biolayer interferometry was used to obtain the affinity constant of an aptamer for lipopolysaccharide, which had an equilibrium dissociation constant of 22.9 nM. The amine-terminated aptamer was then assembled on a gold electrode surface using 3-mercaptopropionic acid as an intermediate linker. The modification of the gold electrode was confirmed by cyclic voltammetry and electrochemical impedance spectroscopy. In the range of 0.001-1 EU/mL, the increase in electron transfer resistance of the biosensor was linear with the logarithmic value of the endotoxin concentration. The constructed biosensor exhibits sensitivity and a low limit of detection.

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“…The light and electrochemical sensors have also attracted increasing attention in endotoxin detection lately. Aptamers, , binding proteins, , and polymyxin B were used as endotoxin recognition receptors and probes to achieve high specificity and rapid response of endotoxin. Then, biological signals are converted into fluorescence, − chemiluminescence, and electrical ones , by sensors to complete the detection.…”

Section: Introductionmentioning

confidence: 99%

In Situ Detection of Endotoxin in Bacteriostatic Process by SERS Chip Integrated Array Microchambers within Bioscaffold Nanostructures and SERS Tags

Xiang

et al. 2020

ACS Appl. Mater. Interfaces

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In order to achieve real-time and in situ detection of endotoxin, which is an important and significant clinical test index, surface-enhanced Raman spectroscopy (SERS) chip integrated array microchambers within bioscaffold nanostructures and a SERS monitoring strategy were proposed in this paper. After sputtering of nanogold on the cicada wing, which was selected as a natural template, and polydimethylsiloxane bonding, array-type chambers within bioscaffold nanostructures were prepared for in situ bacterial culture and monitoring of endotoxin in the bacteriostasis process by SERS. Meanwhile, the SERS tag modified with the DNA aptamer was prepared and added into this complex biochemical reaction to further improve the sensitivity and selectivity. A new method for in situ detection of endotoxin was thus established. The detection time was shortened to 100 s, and the detection limit was as low as 6.25 ng/mL. Pseudomonas aeruginosa was cultured in situ in the chamber of the SERS chip with antimicrobial agents in 0−72 h. The endotoxin released in the antibacterial process was monitored by the designed SERS detection strategy. The results obtained by SERS analysis were consistent with those of the ELISA kit.

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Selecting DNA aptamers for endotoxin separation

Abstract: Using these aptamer-magnetic beads, a new method to separate endotoxin was developed to enable specific separation of endotoxin that can be applied to drug and food products.

Cited by 5 publications

References 11 publications

Synthesis of fluorescence labelled aptamers for use as low‑cost reagents in HIV/AIDS research and diagnostics

Synthesis of fluorescence labelled aptamers for use as low‑cost reagents in HIV/AIDS research and diagnostics

Construction and application of an electrochemical biosensor based on an endotoxin aptamer

In Situ Detection of Endotoxin in Bacteriostatic Process by SERS Chip Integrated Array Microchambers within Bioscaffold Nanostructures and SERS Tags

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