RNA aptamers with specificity for heparosan and chondroitin glycosaminoglycans
Author
Kizer, Megan; Li, Peiqin; Cress, Brady F.; Lin, Lei; Jing, Tom T.; Zhang, Xing; Xia, Ke; Linhardt, Robert J.; Wang, XingOther Contributors
Date Issued
2018-10-10Subject
Biology; Chemistry and chemical biology; Chemical and biological engineering; Biomedical engineeringDegree
Terms of Use
CC BY-NC-ND : this license allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.; Attribution-NonCommercial-NoDerivs 3.0 United StatesFull Citation
RNA aptamers with specificity for heparosan and chondroitin glycosaminoglycans, M. Kizer, P. Li, B. F. Cress, L. Lin, T. T. Jing, X. Zhang, R. J. Linhardt, X. Wang, ACS Omega, 3, 13667−13675, 2018.Metadata
Show full item recordAbstract
In this study, two respective groups of RNA aptamers have been selected against two main classes of glycosaminoglycans (GAGs), heparosan, and chondroitin, as they have proven difficult to specifically detect in biological samples. GAGs are linear, anionic, polydisperse polysaccharides found ubiquitously in nature, yet their detection remains problematic. GAGs comprised repeating disaccharide units, consisting of uronic acid and hexosamine residues that are often also sulfated at various positions. Monoclonal antibodies are frequently used in biology and medicine to recognize various biological analytes with high affinity and specificity. However, GAGs are conserved across the whole animal phylogenic tree and are nonimmunogenic in hosts traditionally used for natural antibody generation. Thus, it has been challenging to obtain high affinity, selective antibodies that recognize various GAGs. In the absence of anti-GAG antibodies, glycobiologists have relied on the use of specific enzymes to convert GAGs to oligosaccharides for analysis by mass spectrometry. Unfortunately, while these methods are sensitive, they can be labor-intensive and cannot be used for in situ detection of intact GAGs in cells and tissues. Aptamers are single-stranded oligonucleotide (DNA or RNA) ligands capable of high selectivity and high affinity detection of biological analytes. Aptamers can be developed in vitro by the systematic evolution of ligands by exponential enrichment (SELEX) to recognize nonimmunogenic targets, including neutral carbohydrates. This study utilizes the SELEX method to generate RNA aptamers, which specifically bind to the unmodified GAGs, heparosan, and chondroitin. Binding confirmation and cross-screening with other GAGs were performed using confocal microscopy to afford three specific GAGs to each target. Affinity constant of each RNA aptamer was obtained by fluorescent output after interaction with the respective GAG target immobilized on plates; the KD values were determined to be 0.71–1.0 μM for all aptamers. Upon the success of chemical modification (to stabilize RNA aptamers in actual biological systems) and fluorescent tagging (to only visualize RNA aptamers) of these aptamers, they would be able to serve as a specific detection reagent of these important GAGs in biological samples.;Description
ACS Omega, 3, 13667−13675; Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.Department
The Linhardt Research Labs.; The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS);Publisher
American Chemical Society (ACS)Relationships
The Linhardt Research Labs Online Collection; Rensselaer Polytechnic Institute, Troy, NY; ACS Omega; https://harc.rpi.edu/;Access
CC BY-NC-ND — Creative Commons Attribution-NonCommercial-NoDerivatives; A full text version is available in DSpace@RPI; Open Access;Collections
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