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Immobilization of Thrombin to Alginate Gel and in vitro Application of Immobilized Thrombin

Year 2017, Volume: 45 Issue: 3, 287 - 294, 01.09.2017

Abstract

Bleeding is one of the biggest problems of humanity. Blood loss in road traffic accidents and surgical operations is an important cause of death. In this study, a product which could decrease bleeding time was developed. The production process of developed product was very simple and the product has commercialization potential. Thrombin was immobilized to calcium alginate beads via ionic interactions. In this method, negative charged carboxylate groups of calcium alginate interacted ionically with the positive charge of thrombin. Optimization studies of thrombin immobilization were performed for increasing immobilization efficiency and it was found to be 4.1% at the end of optimization studies. The thrombin immobilized calcium alginate beads were used for the recalcification time test. The recalcification time is an indicator of bleeding time. The thrombin immobilized calcium alginate lowered recalcification time to 50 percent. Developed product has a potential to stop bleeding.

References

  • K.H. Koo, M.A. Mont, L.C. Jones, Osteonecrosis, (L.C. Jones and D. McK. Ciombor, Osteonecrosis and Intravascular Coagulation Revisited). Berlin: Heidelberg Springer, 2014.
  • P. Wang, Y. Song, M.D. Weir, J. Suna, L. Zhao, C.G. Simond, H.H.K. Xu, A self-setting iPSMSC-alginate- calcium phosphate paste for bone tissue engineering, Dent. Mater., 32 (2016) 252-263.
  • A. Ilie, C. Ghiulică, E. Andronescu, A. Cucuruz, A. Ficai, New composite materials based on alginate and hydroxyapatite as potential carriers for ascorbic acid, Int. J. Pharm., 510 (2016) 501-507.
  • A Akkaya, A.H. Uslan, Sequential immobilization of urease to glycidyl methacrylate grafted sodium alginate, J. Mol. Catal. B-Enzym., 67 (2010) 195-201.
  • D. Kim, T.S. Lee, The detection of thrombin using a mixture of a fluorescent conjugated polyelectrolyte and fibrinogen and implementation of a logic gate, Chem. Commun., 50 (2014) 5833-5836.
  • A. Akkaya, N.K. Pazarlioglu, Thrombin immobilization to poly(methacrylicacid) graft polymerized PET and PAN fabrics, Fiber. Polym., 14 (2013) 358-364.
  • A. Akkaya, N.K. Pazarlioglu, Thrombin immobilization to enzymatic modified PET and PAN fabrics and their applications, Fiber. Polym., 13 (2012) 985-993.
  • R. Thakur, A. Kumar, B. Bose, D. Panda, D. Saikia, P. Chattopadhyay, A.K. Mukherjee, A new peptide (Ruviprase) purified from the venom of Daboia russelii russelii shows potent anticoagulant activity via non-enzymatic inhibition of thrombin and factor Xa, Biochimie, 105 (2014) 149-158.
  • A.V. Sokolov, L. Acquasaliente, V.A. Kostevich, R. Frasson, E.T. Zakharova, G. Pontarollo, V. De Filippis, Thrombin inhibits the anti-myeloperoxidase and ferroxidase functions of ceruloplasmin: relevance in rheumatoid arthritis, Free Radical Bio. Med., 86 (2015) 279-294.
  • T. Xu, A. Lew-Tabor, M. Rodriguez-Valle, Effective inhibition of thrombin by Rhipicephalus microplus serpin-15 (RmS-15) obtained in the yeast Pichia pastoris, Ticks Tick Borne Dis., 7 (2016) 180-187.
  • D.E. Vivas-Ruiz, G.A. Sandoval, J. Mendoza, R.R. Inga, S. Gontijo, M. Richardson, E.F. Sanchez, Coagulant thrombin-like enzyme (barnettobin) from Bothrops barnetti venom: molecular sequence analysis of its cDNA and biochemical properties, Biochimie, 95 (2013) 1476-1486.
  • M.T. Cargnelutti, A.F. Marques, D. Esser, R.Q. Monteiro, M.U. Kassack, L.M.T. Lima, Allosteric activation of human α-thrombin through exosite 2 by suramin analogs, Arch. Biochem. Biophys., 520 (2012) 36-41.
  • A. Akkaya, N.K. Pazarlioglu, Thrombin immobilization to methacrylic acid grafted poly(3-hydroxybutyrate) and its in vitro application, Prep. Biochem. Biotech., 43 (2013) 48-59.
  • M.F. Elahi, G. Guan, L. Wang, Hemocompatibility of surface modified silk fibroin materials: A Review, Rev. Adv. Mater. Sci., 38 (2014) 148-159.
  • A.H.P. Gracher, A.G. Santana, T.R. Cipriani, M. Iacomini, A procoagulant chemically sulfated mannan, Carbohyd. Polym., 136 (2016) 177-186.
  • X. Xu, W. Liu, W. Li, S. Liu, Anticoagulant activity of crude extract of Holotrichia diomphalia larvae, J. Ethnopharmacol., 177 (2016) 28-34.
  • X. Wanga, L. Hu, C. Li, L. Gan, M. He, X. Hea, W. Tian, M. Li, L. Xu, Y. Li, Y. Chen, Improvement in physical and biological properties of chitosan/soyprotein films by surface grafted heparin, Int. J. Biol. Macromol., 83 (2016) 19-29.
  • J. Li, S.Y. Kim, X. Chen, H.J. Park, Calcium-alginate beads loaded with gallic acid: Preparation and characterization, LWT-Food Sci. Technol., 68 (2016) 667-673.
  • Y. Takata, R. Shimohigoshi, K. Yamamoto, J-I. Kadokawa, Enzymatic synthesis of dendritic amphoteric α-glucans by thermostable phosphorylase catalysis, Macromol. Biosci., 14 (2014) 1437-1443.
  • A. Imeson, Food Stabilisers, Thickeners and Gelling Agents, UK. FMC BioPolymer, 2010.
  • de la Escosura-Muñiz, W. Chunglok, W. Surareungchai, A. Merkoçi, Nanochannels for diagnostic of thrombin-related diseases in human blood, Biosens. Bioelectron., 40 (2013) 24-31.
  • M. Babaei, A. Karimi, M.A. Hejazi, Use of mesoporous MnO2 as a support for immobilization of lipase from Candida rugose, Chem. Ind. Chem. Eng., 20 (2014) 371-378.
  • M. Elias, G. Wieczorek, S. Rosenne, D.S. Tawfik, The universality of enzymatic rate-temperature dependency, Trends Biochem. Sci., 39 (2014) 1-7.
  • A. Gao, F. Liu, L. Xue, Preparation and evaluation of heparin-immobilized poly(lacticacid) (PLA) membrane for hemodialysis, J. Membrane Sci., 452 (2014) 390-399.

Trombinin Alginat Jeline İmmobilizasyonu ve İmmobilize Trombinin In Vitro Uygulaması

Year 2017, Volume: 45 Issue: 3, 287 - 294, 01.09.2017

Abstract

Kanama, insanlığın en büyük problemlerinden biridir. Trafik kazaları ve cerrahi operasyonlardaki kan kaybı, ölümlerin önemli bir sebebidir. Bu çalışmada, kanama süresini azaltabilecek bir ürün geliştirildi. Geliştirilen ürünün üretim süreci çok basittir. Ayrıca ürünün ticarileşme potansiyeli bulunmaktadır. Trombin, iyonik etkileşimler yoluyla kalsiyum aljinat boncuklara immobilize edildi. Bu yöntemde, kalsiyum aljinatın negatif yüklü karboksilat grupları, trombinin pozitif yükleriyle iyonik olarak etkileşime girdi. İmmobilizasyon verimini arttırmak için trombin immobilizasyonunun optimizasyon çalışmaları yapıldı. Optimizasyon çalışmalarının sonunda immobilizasyon verimi %4.1 olarak bulundu. Trombin immobilize kalsiyum aljinat boncukları, rekalsifikasyon zamanı testi için kullanıldı. Rekalsifikasyon zamanı, kanama süresi için bir göstergedir. Trombin immobilize kalsiyum aljinat, rekalsifikasyon zamanını %50’ye düşürdü. Geliştirilen ürün kanamayı durdurma potansiyeline sahiptir

References

  • K.H. Koo, M.A. Mont, L.C. Jones, Osteonecrosis, (L.C. Jones and D. McK. Ciombor, Osteonecrosis and Intravascular Coagulation Revisited). Berlin: Heidelberg Springer, 2014.
  • P. Wang, Y. Song, M.D. Weir, J. Suna, L. Zhao, C.G. Simond, H.H.K. Xu, A self-setting iPSMSC-alginate- calcium phosphate paste for bone tissue engineering, Dent. Mater., 32 (2016) 252-263.
  • A. Ilie, C. Ghiulică, E. Andronescu, A. Cucuruz, A. Ficai, New composite materials based on alginate and hydroxyapatite as potential carriers for ascorbic acid, Int. J. Pharm., 510 (2016) 501-507.
  • A Akkaya, A.H. Uslan, Sequential immobilization of urease to glycidyl methacrylate grafted sodium alginate, J. Mol. Catal. B-Enzym., 67 (2010) 195-201.
  • D. Kim, T.S. Lee, The detection of thrombin using a mixture of a fluorescent conjugated polyelectrolyte and fibrinogen and implementation of a logic gate, Chem. Commun., 50 (2014) 5833-5836.
  • A. Akkaya, N.K. Pazarlioglu, Thrombin immobilization to poly(methacrylicacid) graft polymerized PET and PAN fabrics, Fiber. Polym., 14 (2013) 358-364.
  • A. Akkaya, N.K. Pazarlioglu, Thrombin immobilization to enzymatic modified PET and PAN fabrics and their applications, Fiber. Polym., 13 (2012) 985-993.
  • R. Thakur, A. Kumar, B. Bose, D. Panda, D. Saikia, P. Chattopadhyay, A.K. Mukherjee, A new peptide (Ruviprase) purified from the venom of Daboia russelii russelii shows potent anticoagulant activity via non-enzymatic inhibition of thrombin and factor Xa, Biochimie, 105 (2014) 149-158.
  • A.V. Sokolov, L. Acquasaliente, V.A. Kostevich, R. Frasson, E.T. Zakharova, G. Pontarollo, V. De Filippis, Thrombin inhibits the anti-myeloperoxidase and ferroxidase functions of ceruloplasmin: relevance in rheumatoid arthritis, Free Radical Bio. Med., 86 (2015) 279-294.
  • T. Xu, A. Lew-Tabor, M. Rodriguez-Valle, Effective inhibition of thrombin by Rhipicephalus microplus serpin-15 (RmS-15) obtained in the yeast Pichia pastoris, Ticks Tick Borne Dis., 7 (2016) 180-187.
  • D.E. Vivas-Ruiz, G.A. Sandoval, J. Mendoza, R.R. Inga, S. Gontijo, M. Richardson, E.F. Sanchez, Coagulant thrombin-like enzyme (barnettobin) from Bothrops barnetti venom: molecular sequence analysis of its cDNA and biochemical properties, Biochimie, 95 (2013) 1476-1486.
  • M.T. Cargnelutti, A.F. Marques, D. Esser, R.Q. Monteiro, M.U. Kassack, L.M.T. Lima, Allosteric activation of human α-thrombin through exosite 2 by suramin analogs, Arch. Biochem. Biophys., 520 (2012) 36-41.
  • A. Akkaya, N.K. Pazarlioglu, Thrombin immobilization to methacrylic acid grafted poly(3-hydroxybutyrate) and its in vitro application, Prep. Biochem. Biotech., 43 (2013) 48-59.
  • M.F. Elahi, G. Guan, L. Wang, Hemocompatibility of surface modified silk fibroin materials: A Review, Rev. Adv. Mater. Sci., 38 (2014) 148-159.
  • A.H.P. Gracher, A.G. Santana, T.R. Cipriani, M. Iacomini, A procoagulant chemically sulfated mannan, Carbohyd. Polym., 136 (2016) 177-186.
  • X. Xu, W. Liu, W. Li, S. Liu, Anticoagulant activity of crude extract of Holotrichia diomphalia larvae, J. Ethnopharmacol., 177 (2016) 28-34.
  • X. Wanga, L. Hu, C. Li, L. Gan, M. He, X. Hea, W. Tian, M. Li, L. Xu, Y. Li, Y. Chen, Improvement in physical and biological properties of chitosan/soyprotein films by surface grafted heparin, Int. J. Biol. Macromol., 83 (2016) 19-29.
  • J. Li, S.Y. Kim, X. Chen, H.J. Park, Calcium-alginate beads loaded with gallic acid: Preparation and characterization, LWT-Food Sci. Technol., 68 (2016) 667-673.
  • Y. Takata, R. Shimohigoshi, K. Yamamoto, J-I. Kadokawa, Enzymatic synthesis of dendritic amphoteric α-glucans by thermostable phosphorylase catalysis, Macromol. Biosci., 14 (2014) 1437-1443.
  • A. Imeson, Food Stabilisers, Thickeners and Gelling Agents, UK. FMC BioPolymer, 2010.
  • de la Escosura-Muñiz, W. Chunglok, W. Surareungchai, A. Merkoçi, Nanochannels for diagnostic of thrombin-related diseases in human blood, Biosens. Bioelectron., 40 (2013) 24-31.
  • M. Babaei, A. Karimi, M.A. Hejazi, Use of mesoporous MnO2 as a support for immobilization of lipase from Candida rugose, Chem. Ind. Chem. Eng., 20 (2014) 371-378.
  • M. Elias, G. Wieczorek, S. Rosenne, D.S. Tawfik, The universality of enzymatic rate-temperature dependency, Trends Biochem. Sci., 39 (2014) 1-7.
  • A. Gao, F. Liu, L. Xue, Preparation and evaluation of heparin-immobilized poly(lacticacid) (PLA) membrane for hemodialysis, J. Membrane Sci., 452 (2014) 390-399.
There are 24 citations in total.

Details

Primary Language English
Journal Section Research Article
Authors

Alper Akkaya This is me

Publication Date September 1, 2017
Published in Issue Year 2017 Volume: 45 Issue: 3

Cite

APA Akkaya, A. (2017). Immobilization of Thrombin to Alginate Gel and in vitro Application of Immobilized Thrombin. Hacettepe Journal of Biology and Chemistry, 45(3), 287-294.
AMA Akkaya A. Immobilization of Thrombin to Alginate Gel and in vitro Application of Immobilized Thrombin. HJBC. September 2017;45(3):287-294.
Chicago Akkaya, Alper. “Immobilization of Thrombin to Alginate Gel and in Vitro Application of Immobilized Thrombin”. Hacettepe Journal of Biology and Chemistry 45, no. 3 (September 2017): 287-94.
EndNote Akkaya A (September 1, 2017) Immobilization of Thrombin to Alginate Gel and in vitro Application of Immobilized Thrombin. Hacettepe Journal of Biology and Chemistry 45 3 287–294.
IEEE A. Akkaya, “Immobilization of Thrombin to Alginate Gel and in vitro Application of Immobilized Thrombin”, HJBC, vol. 45, no. 3, pp. 287–294, 2017.
ISNAD Akkaya, Alper. “Immobilization of Thrombin to Alginate Gel and in Vitro Application of Immobilized Thrombin”. Hacettepe Journal of Biology and Chemistry 45/3 (September 2017), 287-294.
JAMA Akkaya A. Immobilization of Thrombin to Alginate Gel and in vitro Application of Immobilized Thrombin. HJBC. 2017;45:287–294.
MLA Akkaya, Alper. “Immobilization of Thrombin to Alginate Gel and in Vitro Application of Immobilized Thrombin”. Hacettepe Journal of Biology and Chemistry, vol. 45, no. 3, 2017, pp. 287-94.
Vancouver Akkaya A. Immobilization of Thrombin to Alginate Gel and in vitro Application of Immobilized Thrombin. HJBC. 2017;45(3):287-94.

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