Cost-effective indirect ELISA method for determination of recombinant human erythropoietin in production streams

  • Tomáš Molnár
  • Mária Bartošová
  • Monika Antošová
  • Ľudovít Škultéty
  • Milan Polakovič
Original Paper


Effective development of production of any therapeutic protein requires a relatively fast, reliable, and economical method of target protein monitoring in the outlet streams of cultivation and separation steps. The aim of this work was to develop a method for the determination of recombinant human erythropoietin (rhEPO) produced by human embryonic kidney (HEK) cells. An indirect ELISA method was designed that meets all requirements to fulfill this goal. It was found that an HEK cell-cultivation medium, containing plant hydrolyzate, is a suitable coating buffer, which solves the problem of competitive binding of byproducts. The method has a linear range of 0.078–4 μg/mL allowing accurate determination of rhEPO in relatively concentrated solutions (about 200 μg/mL) without excessive dilution. For the multicomponent rhEPO concentrate at the 400-fold dilution, the inter-assay coefficient of variation was found to be 3.3%. The cost analysis showed that the designed indirect ELISA assay is by one–two orders of magnitude cheaper than the existing commercial sandwich ELISA kits.


Recombinant human erythropoietin Protein determination Indirect ELISA Immunoassay Cost analysis 



This work was supported by the grants from the Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic for Structural Funds of EU (Grant no: ITMS 26240220071), the Slovak Research and Development Agency (Grant no: APVV-14-0474), and the Slovak Grant Agency for Science (Grant no: VEGA 1/0573/17).

Compliance with ethical standards

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.


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Copyright information

© Institute of Chemistry, Slovak Academy of Sciences 2019

Authors and Affiliations

  1. 1.Department of Chemical and Biochemical Engineering, Institute of Chemical and Environmental Engineering, Faculty of Chemical and Food TechnologySlovak University of TechnologyBratislavaSlovakia
  2. 2.Institute of Virology, Biomedical Research CenterSlovak Academy of SciencesBratislavaSlovakia

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