ISSN print edition: 0366-6352
ISSN electronic edition: 1336-9075
Registr. No.: MK SR 9/7
Cost-effective indirect ELISA method for determination of recombinant human erythropoietin in production streams
Tomáš Molnár, Mária Bartošová, Monika Antošová, Ľudovít Škultéty, and Milan Polakovič
Slovak University of Technology, Bratislava, Slovakia
Abstract: 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.
Keywords: Recombinant human erythropoietin ; Protein determination ; Indirect ELISA ; Immunoassay ; Cost analysis
Full paper is available at www.springerlink.com.
Chemical Papers 73 (3) 713–718 (2019)
Saturday, October 16, 2021