title: Evaluation of an in vitro cell culture assay for the potency assessment of recombinant human erythropoietin.
abstract: Recombinant human erythropoietin is a sialoglycoprotein that stimulates erythropoiesis. To assess potency of human erythropoietin produced by recombinant technology, we investigated an in vitro TF-1 cell proliferation assay, which was applied in conjunction with a reversed-phase liquid chromatography method for the determination of the content of sialic acids. The results obtained, which were higher than 126.8ng/μg, were compared with those obtained with the in vivo normocythaemic mouse bioassay. The in vitro assay resulted in a non-significant lower mean difference of the estimated potencies (0.61% ± 0.026, p > 0.05). The use of this combination of methods represents an advance toward the establishment of alternative in vitro approaches, in the context of the Three Rs, for the potency assessment of biotechnology-derived medicines.
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria-RS, Brazil.
authors.forename: Francine T
authors.initials: FT
authors.lastname: Machado
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria-RS, Brazil.
authors.forename: Fernanda P S
authors.initials: FP
authors.lastname: Maldaner
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria-RS, Brazil.
authors.forename: Rafaela F
authors.initials: RF
authors.lastname: Perobelli
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria-RS, Brazil.
authors.forename: Bruna
authors.initials: B
authors.lastname: Xavier
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria-RS, Brazil.
authors.forename: Francielle S
authors.initials: FS
authors.lastname: da Silva
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria-RS, Brazil.
authors.forename: Guilherme W
authors.initials: GW
authors.lastname: de Freitas
authors.affiliation: Biotechnology Department, IPEN-CNEN, São Paulo-SP, Brazil.
authors.forename: Paolo
authors.initials: P
authors.lastname: Bartolini
authors.affiliation: Biotechnology Department, IPEN-CNEN, São Paulo-SP, Brazil.
authors.forename: M Tereza C P
authors.initials: MT
authors.lastname: Ribela
authors.affiliation: Biotechnology Department, IPEN-CNEN, São Paulo-SP, Brazil.
authors.forename: Sérgio L
authors.initials: SL
authors.lastname: Dalmora
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D004921:Erythropoietin / Q000494:pharmacology*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000494:pharmacology
chemical_list: D011994:Recombinant Proteins; D004921:Erythropoietin