Selection and Identification of Novel Aptamers Specific for Clenbuterol Based on ssDNA Library Immobilized SELEX and Gold Nanoparticles Biosensor

Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry
Xixia LiuYaoyao Hou

Abstract

We describe a multiple combined strategy to discover novel aptamers specific for clenbuterol (CBL). An immobilized ssDNA library was used for the selection of specific aptamers using the systematic evolution of ligands by exponential enrichment (SELEX). Progress was monitored using real-time quantitative PCR (Q-PCR), and the enriched library was sequenced by high-throughput sequencing. Candidate aptamers were picked and preliminarily identified using a gold nanoparticles (AuNPs) biosensor. Bioactive aptamers were characterized for affinity, circular dichroism (CD), specificity and sensitivity. The Q-PCR amplification curve increased and the retention rate was about 1% at the eighth round. Use of the AuNPs biosensor and CD analyses determined that six aptamers had binding activity. Affinity analysis showed that aptamer 47 had the highest affinity (Kd = 42.17 ± 8.98 nM) with no cross reactivity to CBL analogs. Indirect competitive enzyme linked aptamer assay (IC-ELAA) based on a 5'-biotin aptamer 47 indicated the limit of detection (LOD) was 0.18 ± 0.02 ng/L (n = 3), and it was used to detect pork samples with a mean recovery of 83.33⁻97.03%. This is the first report of a universal strategy including library fixation, Q-PCR monit...Continue Reading

References

Oct 25, 1995·Nucleic Acids Research·K P Williams, D P Bartel
Apr 27, 2007·Analytical Chemistry·Wolfgang HaissDavid G Fernig
Feb 26, 2008·Journal of the American Chemical Society·Weian ZhaoYingfu Li
Feb 11, 2011·Journal of Agricultural and Food Chemistry·Jiang HeXianjin Liu
Dec 11, 2013·The Analyst·Jeroen VanbrabantLuc Michiels
Jan 28, 2014·Journal of Pharmaceutical and Biomedical Analysis·Nina MencinBorut Šolar
Jul 30, 2014·Chemical Communications : Chem Comm·Van-Thuan NguyenMan Bock Gu
Aug 28, 2015·Journal of Photochemistry and Photobiology. B, Biology·M M V RamanaSachin Pardeshi
May 26, 2016·Biotechnology and Applied Biochemistry·Mohammad HeiatGholamreza Farnoosh
Jan 7, 2017·Journal of Analytical Methods in Chemistry·Rui-Yun TianShi-Ying Lu

❮ Previous
Next ❯

Citations

Jan 21, 2020·Journal of Biological Engineering·Qingxiu LiuJun Sheng Lin
Feb 14, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Qi LuSirui Chen
Mar 27, 2020·International Journal of Molecular Sciences·Wei ZhangJun Sheng Lin
Jun 5, 2021·Applied Biochemistry and Biotechnology·Hongli FanZhuoya Luo

❮ Previous
Next ❯

Methods Mentioned

BETA
SELEX
biosensors
biosensor
circular dichroism
enzyme-linked aptamer assay
PCR
electrophoresis

Software Mentioned

GraphPad
mfold
clustal
Graphpad Prism
clustalX2
TreeView

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.