Cell-Specific Aptamer Characterization

Chen, Tao; Wu, Cuichen; Wang, Tan

doi:10.1007/978-3-662-46226-3_4

Cited by 1 publication

(1 citation statement)

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“…They display very high target specificity and superior binding affinity for the target molecule with dissociation constants from nano to picomolar range and other features that make them more appealing than antibodies [ 6 ]. Since first reported in the 1990s, several SELEX methods have been developed to facilitate more efficient isolation of aptamers for a wide variety of targets including ions [ 7 , 8 ], small molecules [ 9 ], proteins [ 10 ], and whole cells [ 11 ]. Graphene oxide-assisted SELEX (GO-SELEX) is an immobilization-free method that can be easily implemented to enrich the aptamer pool [ 12 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

Isolation and Characterization of a ssDNA Aptamer against Major Soluble Antigen of Renibacterium salmoninarum

et al. 2022

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Bacterial kidney disease (BKD) is a major health problem of salmonids, affecting both wild and cultured salmon. The disease is caused by Renibacterium salmoninarum (Rs), a fastidious, slow-growing and strongly Gram-positive diplobacillus that produces chronic, systemic infection characterized by granulomatous lesions in the kidney and other organs, often resulting in death. Fast detection of the pathogen is important to limit the spread of the disease, particularly in hatcheries or aquaculture facilities. Aptamers are increasingly replacing conventional antibodies as platforms for the development of rapid diagnostic tools. In this work, we describe the first instance of isolating and characterizing a ssDNA aptamer that binds with high affinity to p57 or major soluble antigen (MSA), the principal antigen found on the cell wall surface of Rs. Specifically, in this study a construct of the full-length protein containing a DNA binding domain (MSA-R2c) was utilized as target. Aptamers were isolated from a pool of random sequences using GO-SELEX (graphene oxide-systematic evolution of ligands by exponential enrichment) protocol. The selection generated multiple aptamers with conserved motifs in the random region. One aptamer with high frequency of occurrence in different clones was characterized and found to display a strong binding affinity to MSA-R2c with a Kd of 3.0 ± 0.6 nM. The aptamer could be potentially utilized for the future development of a sensor for rapid and onsite detection of Rs in water or in infected salmonids, replacing time-consuming and costly lab analyses.

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