A Monoclonal Antibody Against Bovine Thrombin Reacting to the C-Terminal Side of Thrombin

Moriyama, Tomoe; Nakamura, Mariko; Kinjoh, Kiyohiko; Tanaka, Tatsuo; Kosugi, Tadayoshi

doi:10.1089/15368590152740806

Cited by 2 publications

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References 40 publications

(31 reference statements)

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“…Since then topical bovine thrombin and fibrin sealants are extensively used in cardiovascular surgery, otolaryngologic surgery and neurosurgery (Brennan, 1991). There is 85% homology between bovine thrombin and human thrombin, so most antibodies against bovine thrombin cross-react with human thrombin (Moriyama et al, 2001). Rare antibodies against bovine thrombin but not cross-reacting with human thrombin have been reported.…”

Section: Introductionmentioning

confidence: 99%

RNA aptamers specific for bovine thrombin

Liu¹,

Zhang²,

Cao³

et al. 2003

J of Molecular Recognition

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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 nM, 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.

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Section: Introductionmentioning

confidence: 99%

RNA aptamers specific for bovine thrombin

Liu¹,

Zhang²,

Cao³

et al. 2003

J of Molecular Recognition

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“…As the quantity of bound thrombin generated in vivo would be too small to be isolated and identified. We tried to detect it using a monoclonal antibody [36].…”

mentioning

confidence: 99%

Bound Thrombin from Crushed Clots Is Composed of α-Thrombin and the N-Terminal Regions of α- and γ-Chains of Fibrinogen

Kinjoh

Nakamura²,

Zeng³

et al. 2002

Pathophysiol Haemos Thromb

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We aimed at clarifying the structural characteristics of the bound thrombin that is liberated by mechanical breakdown of fibrin clots. Fibrin clots were prepared with bovine thrombin and rabbit fibrinogen, and were crushed mechanically with a glass rod. The supernatant of the crushed clots was subjected to immunoaffinity chromatography to isolate the bound thrombin. Western blotting analysis revealed that the bound thrombin could be reacted with both antithrombin and antifibrinogen under unreduced conditions. SDS-PAGE under reduced conditions revealed that there were three bands, two of which were found to be the N-terminal fragments of the α- and γ-chains of fibrinogen. The bound thrombin could be dissociated into three distinct fibrin fragments and bovine α-thrombin when denatured by 8 M urea. Thus, the bound thrombin liberated from crushed clots is a stable complex between bovine α-thrombin and fibrin fragments of the N-terminal regions of rabbit α- and γ-chains.

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A Monoclonal Antibody Against Bovine Thrombin Reacting to the C-Terminal Side of Thrombin

Cited by 2 publications

References 40 publications

RNA aptamers specific for bovine thrombin

RNA aptamers specific for bovine thrombin

Bound Thrombin from Crushed Clots Is Composed of α-Thrombin and the N-Terminal Regions of α- and γ-Chains of Fibrinogen

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