The development of a method for determining the biological activity of erythropoietin preparations in vitro
Abstract
The method for the assessment of biological activity of erythropoietin preparations in vitro on TF-1 sensitive cells culture has been developed. The special characteristics of measurements and calculations of the biological activity of erythropoietin with different levels of glycosylation in vivo and in vitro. Linear response ranges for cell lines to erythropoietin and its hyperglycosylated analogue differ and make (0.39-0.56 ng/ml) and (3.1-12.5 ng/ml), respectively. The results have shown the possibility of using cellular test for comparing and assaying the activity of erythropoietin analogues in standard international units of biological activity (IU) set for erythropoietin.
About the Authors
N. A. Gavrilova
Scientific Centre for Expert Evaluation of Medicinal Products
Russian Federation
Analyst of Office of expertise of antibacterial medical immunobiological preparations of Center for examination and control of medical immunobiological preparations. Candidate of Biological Sciences
Russian Federation
Analyst of Office of expertise of antibacterial medical immunobiological preparations of Center for examination and control of medical immunobiological preparations. Candidate of Biological Sciences
S. A. Cherepushkin
State Research Institute of Selection of Industrial Microorganisms
Russian Federation
Researcher of Department of medical biotechnology
Russian Federation
Researcher of Department of medical biotechnology
N. V. Rykalina
State Research Institute of Selection of Industrial Microorganisms
Russian Federation
Researcher of Laboratory of immunology
Russian Federation
Researcher of Laboratory of immunology
Yu. I. Obukhov
Scientific Centre for Expert Evaluation of Medicinal Products
Russian Federation
Head of Office of expertise of antibacterial medical immunobiological preparations of Center of expertise and control of medical immunobiological preparations
Russian Federation
Head of Office of expertise of antibacterial medical immunobiological preparations of Center of expertise and control of medical immunobiological preparations
References
1. Bunn HF. New agents that stimulate erythropoesis. Blood 2007; 109(3): 868–3.
2. Macdougall IC, Eckardt K-U. Novel strategies for stimulating erythropoiesis and potential treatments for anaemia. Lancet 2006; 368(9539): 947–53. 3. Cossar JD, Lawrence M, Donaldi S. Pegylated erythpoietic compounds. WO2004009627; 2004.
3. Egrie JC, Dwyer E, Browne JK, Hitz A, Lykos MA. Darbepoetin alfa has a longer circulating half-life and greater in vivo potency than recombinant erythropoietin. Exp Hematol. 2003; 31(4): 290–9.
4. Joung CH, Shin JY, Koo JK, Linn JJ, Wang JS, Lee SJ, et al. Production and characterization of long-acting recombinant human albumin-EPO fusion protein expressed in CHO cell. Protein Expr Purif. 2009; 68(2): 137–45.
5. Brines M, Grasso G, Fiordaliso F, Sfacteria A, Ghezzi P, Fratelli M, et al. Erythropoietin mediates tissue protection through an erythropoietin and common beta-subunit heteroreceptor. Proc Natl Acad Sci USA 2004; 101(41): 14907–12.
6. European Pharmacopoeia 8.0/8.4. «Erythropoietin Concentrated Solution».
7. List of European Pharmacopoeia Reference Standards. Available from: http://www.edqm.eu/en/ph-eur-reference-standards-orders-catalogue.
8. Yakovlev AK, Gayderova LA, Alpatova NA, Lobanova TN, Batuashvili TA, Simutenko LV, et al. The stages in standardizing erythropoietin preparations. Biopreparaty 2015; (4): 17–20 (in Russian).
9. Kitamura T, Tange T, Terasawa T, Chiba S, Kuwaki T, Miyagawa K, et al. Establishment and characterization of a unique human cell line that proliferates dependently on GM-CSF, IL-3, or erythropoietin. J Cell Physiol. 1989; 140(2): 323–34.
10. Elliott S, Egrie J, Browne J, Lorenzini T, Busse L, Rogers N, et al. Control of rHuEPO biological activity: the role of carbohydrate. Exp Hematol. 2004; 32(12): 1146–55.
11. Gavrilova NA, Cheremnykh AM, Bobreneva RA, Askerova EV, Kalinina TA, Bulushova NV, et al. The haemopoietic activity and pharmacokinetics of EPO-Fc, EPO-Fcneo and Alb-EPO fused proteins, derivatives of human erythropoietin. Biotehnologiya 2012; (5): 38–49 (in Russian).
12. Cheremnykh AM, Askerova EV, Bobreneva RA, Gavrilova NA, Kalinina TA, Jurin VL. Hybrid protein based on recombinant human erythropoietin, having prolonged action (versions), and method of its production. Patent RF, ¹ 2515914; 2013 (in Russian).
Review
For citations:
Gavrilova N.A., Cherepushkin S.A., Rykalina N.V., Obukhov Yu.I. The development of a method for determining the biological activity of erythropoietin preparations in vitro. BIOpreparations. Prevention, Diagnosis, Treatment. 2016;16(2):120-124. (In Russ.)
Views:
824
Email this article 