Selection and characterization of a DNA aptamer inhibiting coagulation factor XIa View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2017-12

AUTHORS

David A. Donkor, Varsha Bhakta, Louise J. Eltringham-Smith, Alan R. Stafford, Jeffrey I. Weitz, William P. Sheffield

ABSTRACT

Factor XIa (FXIa) is a serine protease that catalyzes the activation of Factor IX (FIX) in the blood coagulation cascade. FXIa and its precursor FXI are emergent therapeutic targets for the development of safer anticoagulant agents. Here, we sought a novel DNA-based agent to inhibit FXIa. Towards this goal, an 80 base, single-stranded DNA aptamer library (containing a 40 base randomized core) was screened for FXIa-binding candidates, using ten rounds of positive and negative selection. After selection, 6 of 89 different sequences inhibited FXIa-mediated chromogenic substrate S2366 cleavage. The most active anti-FXIa aptamer had a hypervariable central sequence 5'-AACCTATCGGACTATTGTTAGTGATTTTTATAGTGT-3' and was designated Factor ELeven Inhibitory APtamer (FELIAP). FELIAP, but not a scrambled aptamer control (SCRAPT), competitively inhibited FXIa-catalyzed S2366 cleavage, FIX activation, and complex formation with antithrombin. No effect of FELIAP on FXI activation was observed. FELIAP inhibited plasma clotting and thrombin generation assays to a significantly greater extent than SCRAPT. Immobilized FELIAP bound FXIa with strong affinity and an equilibrium binding constant (KD) in the low nanomolar range determined using surface plasmon resonance. FELIAP is the first FXIa-inhibitory aptamer to be described and constitutes a lead compound to develop related aptamers for in vivo use. More... »

PAGES

2102

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41598-017-02055-x

DOI

http://dx.doi.org/10.1038/s41598-017-02055-x

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1085436195

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/28522812


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Turtle is a human-readable linked data format.

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RDF/XML is a standard XML format for linked data.

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