학술논문
Potential Application of Recombinant Snake Prothrombin Activator Ecarin in Blood Diagnostics.
Document Type
Academic Journal
Author
Zhao KN; Australian Institute of Biotechnology and Nanotechnology, St Lucia Campus, The University of Queensland, Brisbane, QLD 4072, Australia.; Centre for Kidney Disease Research-Venomics Research, School of Medicine, The University of Queensland, Brisbane, QLD 4072, Australia.; Masci P; Centre for Kidney Disease Research-Venomics Research, School of Medicine, The University of Queensland, Brisbane, QLD 4072, Australia.; Dimeski G; Chemical Pathology, Princess Alexandra Hospital, Woolloongabba, Brisbane, QLD 4102, Australia.; School of Chemistry and Molecular Biosciences, St Lucia Campus, The University of Queensland, Brisbane, QLD 4072, Australia.; Johnson L; Australian Institute of Biotechnology and Nanotechnology, St Lucia Campus, The University of Queensland, Brisbane, QLD 4072, Australia.; Grant M; Q-Sera Pty Ltd., Level 9, 31 Queen St, Melbourne, VIC 3000, Australia.; de Jersey J; School of Chemistry and Molecular Biosciences, St Lucia Campus, The University of Queensland, Brisbane, QLD 4072, Australia.; Lavin MF; Australian Institute of Biotechnology and Nanotechnology, St Lucia Campus, The University of Queensland, Brisbane, QLD 4072, Australia.; Centre for Clinical Research, RBWH Campus, The University of Queensland, Brisbane, QLD 4029, Australia.
Source
Publisher: MDPI Country of Publication: Switzerland NLM ID: 101596414 Publication Model: Electronic Cited Medium: Internet ISSN: 2218-273X (Electronic) Linking ISSN: 2218273X NLM ISO Abbreviation: Biomolecules Subsets: MEDLINE
Subject
Language
English
Abstract
We describe here the purification and cloning of a codon-optimized form of the snake prothrombin activator ecarin from the saw scaled viper ( Echis carinatus ) expressed in mammalian cells. Expression of recombinant ecarin (rEcarin) was carried out in human embryonic kidney cells (HEK) cells under conditions for the development and performance of a novel and scalable recombinant snake ecarin to industry standards. Clotting performance of the rEcarin was established in recalcified citrated whole blood, plasma, and fresh whole blood and found to be comparable to native ecarin (N-Ecarin). Furthermore, hemolysis was observed with N-Ecarin at relatively high doses in both recalcified citrated and fresh whole blood, while clotting was not observed with rEcarin, providing an important advantage for the recombinant form. In addition, rEcarin effectively clotted both recalcified citrated whole blood and fresh whole blood containing different anticoagulants including heparin, warfarin, dabigatran, Fondaparinux, rivaroxaban and apixaban, forming firm clots in the blood collection tubes. These results demonstrate that rEcarin efficiently clots normal blood as well as blood spiked with high concentrations of anticoagulants and has great potential as an additive to blood collection tubes to produce high quality serum for analyte analysis in diagnostic medicine.