Journal of Thrombosis and Thrombolysis

, Volume 35, Issue 1, pp 48–56 | Cite as

The effect of surface contact activation and temperature on plasma coagulation with an RNA aptamer directed against factor IXa

  • Anandi KrishnanEmail author
  • Erwin A. Vogler
  • Bruce A. Sullenger
  • Richard C. Becker


The anticoagulant properties of a novel RNA aptamer that binds FIXa depend collectively on the intensity of surface contact activation of human blood plasma, aptamer concentration, and its binding affinity for FIXa. Accordingly, anticoagulation efficiency of plasma containing any particular aptamer concentration is low when coagulation is strongly activated by hydrophilic surfaces compared to the anticoagulation efficiency in plasma that is weakly activated by hydrophobic surfaces. Anticoagulation efficiency is lower at hypothermic temperatures possibly because aptamer-FIXa binding decreases with decreasing temperatures. Experimental results demonstrating these trends are qualitatively interpreted in the context of a previously established model of anticoagulation efficiency of thrombin-binding DNA aptamers that exhibit anticoagulation properties similar to the FIXa aptamer. In principle, FIXa aptamer anticoagulants should be more efficient and therefore more clinically useful than thrombin-binding aptamers because aptamer binding to FIXa competes only with FX that is at much lower blood concentration than fibrinogen (FI) that competes with thrombin-binding aptamers. Our findings may have translatable relevance in the application of aptamer anticoagulants for clinical conditions in which blood is in direct contact with non-biological surfaces such as those encountered in cardiopulmonary bypass circuits.


Factor IXa inhibition FIXa aptamer Activator surfaces Contact activation Coagulation Recalcification clot time 



Authors would like to thank Dr. Jens Lohrmann and Kristin Bompiani for expert guidance. Anandi Krishnan would also like to thank Dr. Larry Goldstein and the Henrietta B. and Frederick H. Bugher Foundation for postdoctoral fellowship support.


  1. 1.
    Durham SJ, Gold JP (2003). Late complications of cardiac surgery. In: Cohn LH (ed) Cardiac surgery in the adult, Chap 19, 3rd edn. McGraw Hill, New York, pp 535–548Google Scholar
  2. 2.
    Nimjee SM, Rusconi CP, Harrington RA, Sullenger BA (2005) The potential of aptamers as anticoagulants. Trends Cardiovasc Med 15(1):41–45PubMedCrossRefGoogle Scholar
  3. 3.
    Nimjee SM, Rusconi CP, Sullenger BA (2005) Aptamers: an emerging class of therapeutics. Annu Rev Med 56:555–583PubMedCrossRefGoogle Scholar
  4. 4.
    Rusconi CP, Roberts JD, Pitoc GA, Nimjee SM, White RR, Quick G Jr et al (2004) Antidote-mediated control of an anticoagulant aptamer in vivo. Nat Biotechnol 22(11):1423–1428PubMedCrossRefGoogle Scholar
  5. 5.
    Rusconi CP, Scardino E, Layzer J, Pitoc GA, Ortel TL, Monroe D et al (2002) RNA aptamers as reversible antagonists of coagulation factor IXa. Nature 419(6902):90–94PubMedCrossRefGoogle Scholar
  6. 6.
    Sullenger BA, White RR, Rusconi CP (2003) Therapeutic aptamers and antidotes: a novel approach to safer drug design. Ernst Schering Res Found Workshop 43:217–223PubMedGoogle Scholar
  7. 7.
    Becker RC (2005) Cell-based models of coagulation: a paradigm in evolution. J Thromb Thrombolysis 20(1):65–68PubMedCrossRefGoogle Scholar
  8. 8.
    Howard EL, Becker KC, Rusconi CP, Becker RC (2007) Factor IXa inhibitors as novel anticoagulants. Arterioscler Thromb Vasc Biol 27(4):722–727PubMedCrossRefGoogle Scholar
  9. 9.
    Nimjee SM, Keys JR, Pitoc GA, Quick G, Rusconi CP, Sullenger BA (2006) A novel antidote-controlled anticoagulant reduces thrombin generation and inflammation and improves cardiac function in cardiopulmonary bypass surgery. Mol Ther 14(3):408–415PubMedCrossRefGoogle Scholar
  10. 10.
    Becker RC (2007) Emerging paradigms, platforms, and unifying themes in biomarker science. J Am Coll Cardiol 50(18):1777–1780PubMedCrossRefGoogle Scholar
  11. 11.
    Becker RC (2005) Novel constructs for thrombin inhibition. Am Heart J 149(1 Suppl):S61–S72PubMedCrossRefGoogle Scholar
  12. 12.
    Becker RC, Oney S, Becker KC, Rusconi CP, Sullenger B (2007) Nucleic acid aptamers and their complimentary antidotes. Entering an era of antithrombotic pharmacobiologic therapy. Hamostaseologie 27(5):378–382PubMedGoogle Scholar
  13. 13.
    Yavari M, Becker RC (2008) Anticoagulant therapy during cardiopulmonary bypass. J Thromb Thrombolysis 26(3):218–228PubMedCrossRefGoogle Scholar
  14. 14.
    Yavari M, Becker RC (2008) Coagulation and fibrinolytic protein kinetics in cardiopulmonary bypass. J Thromb Thrombolysis 27(1):95–104PubMedCrossRefGoogle Scholar
  15. 15.
    Vogler EA, Siedlecki CA (2009) Contact activation of blood-plasma coagulation. Biomaterials 30(10):1857–1869PubMedCrossRefGoogle Scholar
  16. 16.
    Vogler EA, Nadeau JG, Graper JC (1997) Contact activation of the plasma coagulation cascade. III. Biophysical aspects of thrombin binding anticoagulants. J Biomed Mat Res 40(1):92–103Google Scholar
  17. 17.
    Golas A, Parhi P, Dimachkie ZO, Siedlecki CA, Vogler EA (2010) Surface-energy dependent contact activation of blood factor XII. Biomaterials 31(6):1068–1079PubMedCrossRefGoogle Scholar
  18. 18.
    Vogler EA, Graper JC, Harper GR, Sugg HW, Lander LM, Brittain WJ (1995) Contact activation of the plasma coagulation cascade. I. Procoagulant surface chemistry and energy. J Biomed Mater Res 29(8):1005–1016PubMedCrossRefGoogle Scholar
  19. 19.
    Rusconi CP, Yeh A, Lyerly HK, Lawson JH, Sullenger BA (2000) Blocking the initiation of coagulation by RNA aptamers to factor VIIa. Thromb Haemost 84(5):841–848PubMedGoogle Scholar
  20. 20.
    Povsic TJ, Cohen MG, Chan MY, Zelenkofske SL, Wargin WA, Harrington RA et al (2011) Dose selection for a direct and selective factor IXa inhibitor and its complementary reversal agent: translating pharmacokinetic and pharmacodynamic properties of the REG1 system to clinical trial design. J Thromb Thrombolysis 32(1):21–31PubMedCrossRefGoogle Scholar
  21. 21.
    Povsic TJ, Sullenger BA, Zelenkofske SL, Rusconi CP, Becker RC (2010) Translating nucleic acid aptamers to antithrombotic drugs in cardiovascular medicine. J Cardiovasc Transl Res 3(6):704–716PubMedCrossRefGoogle Scholar
  22. 22.
    Mitropoulos KA (1999) The levels of factor XIIa generated in human plasma on an electronegative surface are insensitive to wide variation in the concentration of FXII, prekallikrein, high molecular weight kininogen or FXI. Thromb Haemost 82(3):1033–1040PubMedGoogle Scholar
  23. 23.
    Mitropoulos KA, Martin JC, Stirling Y, Morrisey JH, Cooper JA (1995) Activation of factors XII and VII induced in citrated plasma in the presence of contact surface. Thromb Res 78(1):67–75PubMedCrossRefGoogle Scholar
  24. 24.
    Vogler EA, Graper JC, Sugg HW, Lander LM, Brittain WJ (1995) Contact activation of the plasma coagulation cascade. II. Protein adsorption to procoagulant surfaces. J Biomed Mater Res 29(8):1017–1028PubMedCrossRefGoogle Scholar
  25. 25.
    Brown B (1973) Hematology principles and procedures. Lea and Febiger, Philadelphia, pp 33–39Google Scholar
  26. 26.
    Vogler EA, Nadeau JG, Graper JC (1998) Contact activation of the plasma coagulation cascade. III. Biophysical aspects of thrombin-binding anticoagulants. J Biomed Mater Res 40(1):92–103PubMedCrossRefGoogle Scholar
  27. 27.
    Zhuo R, Miller R, Bussard KM, Siedlecki CA, Vogler EA (2005) Procoagulant stimulus processing by the intrinsic pathway of blood plasma coagulation. Biomaterials 26(16):2965–2973PubMedCrossRefGoogle Scholar
  28. 28.
    Vogler EA, Graper JC, Harper GR, Lander LM, Brittain WJ (1995) Contact activation of the plasma coagulation cascade. I. Procoagulant surface energy and chemistry. J Biomed Mat Res 29:1005–1016CrossRefGoogle Scholar
  29. 29.
    Zhuo R, Miller R, Bussard KM, Siedlecki CA, Vogler EA (2005) Procoagulant stimulus processing by the intrinsic pathway of blood plasma coagulation. Biomaterials 26:2965–2973PubMedCrossRefGoogle Scholar
  30. 30.
    Zhuo R, Colombo P, Pantano C, Vogler EA (2005) Silicon oxycarbide glasses for blood-contact applications. Acta Biomater 1:583–589PubMedCrossRefGoogle Scholar
  31. 31.
    Anderson NL, Anderson NG (2002) The human plasma proteome: history, character, and diagnostic prospects. Mol Cell Proteomics 1(11):845–867PubMedCrossRefGoogle Scholar
  32. 32.
    Vogler EA (2001) On the origins of water wetting terminology. In: Morra M (ed) Water in biomaterials surface science. John Wiley and Sons, New York, pp 150–182Google Scholar
  33. 33.
    Li Y, Guo L, Zhang F, Zhang Z, Tang J, Xie J (2008) High-sensitive determination of human alpha-thrombin by its 29-mer aptamer in affinity probe capillary electrophoresis. Electrophoresis 29(12):2570–2577PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Anandi Krishnan
    • 1
    Email author
  • Erwin A. Vogler
    • 2
  • Bruce A. Sullenger
    • 3
  • Richard C. Becker
    • 1
    • 3
  1. 1.Duke Clinical Research InstituteDuke University Medical CenterDurhamUSA
  2. 2.Departments of Materials Science & Engineering and BioengineeringThe Pennsylvania State UniversityUniversity ParkUSA
  3. 3.Departments of Surgery and Medicine and Duke Translational Research InstituteDuke University Medical CenterDurhamUSA

Personalised recommendations