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RNA aptamers specific for bovine thrombin
Author(s) -
Liu Xuemei,
Zhang Dajin,
Cao Guojun,
Yang Guang,
Ding Hongmei,
Liu Gang,
Fan Ming,
Shen Beifen,
Shao Ningsheng
Publication year - 2003
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.604
Subject(s) - thrombin , aptamer , systematic evolution of ligands by exponential enrichment , fibrin , chemistry , rna , biochemistry , microbiology and biotechnology , biology , gene , immunology , platelet
Bovine thrombin is widely used in clinical wound healing after surgery. There is 85% homology between bovine thrombin and human thrombin, so most antibodies against bovine thrombin cross‐react with human thrombin. Rare antibodies against bovine thrombin but not cross‐reacting with human thrombin have been reported. RNA ligands (aptamers) have been used to bind to target molecules with sometimes higher specificity than antibodies. Here we report the isolation of aptamers specific for bovine thrombin by systematic evolution of ligands by exponential enrichment (SELEX) from an RNA pool containing a 25‐nucleotide randomized region. After seven rounds of selection, two aptamers specific for bovine thrombin were identified with a K d of 164 and 240 n M , respectively. Significantly, these aptamers do not bind to human thrombin. Secondary structure prediction revealed potential stem‐loop structures for these RNAs. Both RNA aptamers inhibit only bovine thrombin‐catalyzed fibrin clot formation in vitro . Competition assay results suggested that the RNA aptamers might bind to the electropositive domain of bovine thrombin, that is, heparin‐binding site, instead of fibrinogen‐recognition exosite. The resulting bovine‐specific thrombin inhibitor might be used in some clinical applications when bovine thrombin activity needs to be contained or in research where human and bovine thrombin need to be distinguished. Copyright © 2003 John Wiley & Sons, Ltd. Abbreviations used: SELEX systematic evolution of ligands by exponential enrichment

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