title: Stability-indicating capillary zone electrophoresis method for the assessment of recombinant human interleukin-11 and its correlation with reversed-phase liquid chromatography and biossay.
abstract: A stability-indicating capillary zone electrophoresis (CZE) method was validated for the analysis of recombinant human interleukin-11(rhIL-11) using rupatadine fumarate, as internal standard (IS). A fused-silica capillary, (50 µm i.d.; effective length, 40 cm) was used at 25°C; the applied voltage was 20 kV. The background electrolyte solution consisted of 50 mmol L(-1) sodium dihydrogen phosphate solution at pH 3.0. Injections were performed using a pressure mode at 50 mbar for 45 s, with detection by photodiode array detector set at 196 nm. Specificity and stability-indicating capability were established in degradation studies, which also showed that there was no interference of the excipients. The method was linear over the concentration range of 1.0-300 µg mL(-1) (r(2)=0.9992) and the limit of detection (LOD) and limit of quantitation (LOQ) were 0.2 µg mL(-1) and 1.0 µg mL(-1), respectively. The accuracy was 100.4% with bias lower than 1.1%. Moreover, the in vitro cytotoxicity test of the degraded products showed significant differences (p<0.05). The method was applied for the content/potency assessment of rhIL-11 in biopharmaceutical formulations, and the results were correlated to those of a validated reversed-phase LC method (RP-LC) and an TF-1 cell culture assay, showing non-significant differences (p>0.05). In addition the CZE and RP-LC methods were applied for the analysis of rhIL-11 in human plasma. Therefore, the proposed alternative method can be applied to monitor stability, to assure the batch-to-batch consistency and quality of the bulk and finished biotechnology-derived medicine.
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
authors.forename: Ricardo Bizogne
authors.initials: RB
authors.lastname: Souto
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
authors.forename: Fernanda Pavani
authors.initials: FP
authors.lastname: Stamm
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
authors.forename: Jéssica Barbieri
authors.initials: JB
authors.lastname: Schumacher
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
authors.forename: Clovis Dervil Appratto
authors.initials: CD
authors.lastname: Cardoso
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
authors.forename: Guilherme Weber
authors.initials: GW
authors.lastname: de Freitas
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
authors.forename: Rafaela Ferreira
authors.initials: RF
authors.lastname: Perobelli
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil. Electronic address: sdalmora@terra.com.br.
authors.forename: Sérgio Luiz
authors.initials: SL
authors.lastname: Dalmora
mesh_terms: D000818:Animals; D001681:Biological Assay; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D004355:Drug Stability; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D020128:Inhibitory Concentration 50; D017370:Interleukin-11 / Q000032:analysis* / Q000097:blood / Q000494:pharmacology; D055550:Protein Stability; D011994:Recombinant Proteins / Q000032:analysis* / Q000494:pharmacology; D015203:Reproducibility of Results
chemical_list: D017370:Interleukin-11; D011994:Recombinant Proteins
keywords: Biotechnology; Capillary zone electrophoresis; Recombinant human interleukin-11; Reversed-phase liquid chromatography; TF-1 cell culture