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Design of a stable cell line producing recombinant darbepoetin alpha based on CHO cells

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Abstract

A stable cell line that is based on CHO cells and produces 100 mg per liter of culture medium of recombinant darbepoetin alpha with a target glycosylated isoforms effective yield of about 30% has been selected. The expression product of the cell line was characterized and compared to the originator (Aranesp, Amgen). It was shown that the obtained preparation contained all isoforms characteristic of the originator. The created cell line can be used for the development of industrial cultivation and a purification scheme for recombinant darbepoetin alpha production.

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Abbreviations

HPLC:

High Performance Liquid Chromatography

DARB:

darbepoetin alfa

ELISA:

enzyme-linked immunosorbent assay

IEF:

isoelectric focusing

CCF:

cell culture fluid

CZE:

capillary zone electrophoresis

PAGE:

polyacrylamide gel

rhEPO:

recombinant human erythropoietin

EPO:

erythropoietin

CHO :

Chinese hamster ovary cells

CMV:

cytomegalovirus

SDS:

sodium dodecyl sulfate

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Authors and Affiliations

  1. Limited Liability Company “PHARMAPARK,”, Moscow, 117246, Russia

    R. R. Shukurov, N. V. Lobanova, I. N. Savinova, I. G. Vorobyova, A. A. Nurbakov, N. V. Orlova, A. G. Mosina, L. P. Antonova & Yu. A. Seryogin

  2. The State Research Institute for Genetics and Selection of Industrial Microorganisms, Moscow, 117545, Russia

    L. V. Ermolina & R. A. Khamitov

Authors
  1. R. R. Shukurov
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  2. N. V. Lobanova
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  3. I. N. Savinova
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  4. I. G. Vorobyova
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  5. A. A. Nurbakov
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  6. L. V. Ermolina
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  7. N. V. Orlova
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  8. A. G. Mosina
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  9. L. P. Antonova
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  10. R. A. Khamitov
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  11. Yu. A. Seryogin
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Corresponding author

Correspondence to R. R. Shukurov.

Additional information

Original Russian Text © R.R. Shukurov, N.V. Lobanova, I.N. Savinova, I.G. Vorobyova, A.A. Nurbakov, L.V. Ermolina, N.V. Orlova, A.G. Mosina, L.P. Antonova, R.A. Khamitov, Yu.A. Seryogin, 2013, published in Biotekhnologiya, 2013, No. 2, pp. 46–54.

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Shukurov, R.R., Lobanova, N.V., Savinova, I.N. et al. Design of a stable cell line producing recombinant darbepoetin alpha based on CHO cells. Appl Biochem Microbiol 50, 812–818 (2014). https://doi.org/10.1134/S0003683814090063

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  • DOI: https://doi.org/10.1134/S0003683814090063

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