Stability-indicating capillary zone electrophoresis method for the assessment of recombinant human interleukin-11 and its correlation with reversed-phase liquid chromatography and biossay

• Published in: Talanta (Oxford) 2014-06, Vol.123, p.179-185
• Main Authors: Souto, Ricardo Bizogne, Stamm, Fernanda Pavani, Schumacher, Jéssica Barbieri, Cardoso, Clovis Dervil Appratto, de Freitas, Guilherme Weber, Perobelli, Rafaela Ferreira, Dalmora, Sérgio Luiz
• Format: Article
• Language: English
• Subjects: Animals; Biological Assay; Biotechnology; Capillary zone electrophoresis; Cell Line; Cell Line, Tumor; Cell Survival - drug effects; Chromatography, Reverse-Phase - methods; Drug Stability; Electrophoresis, Capillary - methods; Humans; Inhibitory Concentration 50; Interleukin-11 - analysis; Interleukin-11 - blood; Interleukin-11 - pharmacology; Protein Stability; Recombinant human interleukin-11; Recombinant Proteins - analysis; Recombinant Proteins - pharmacology; Reproducibility of Results; Reversed-phase liquid chromatography; TF-1 cell culture
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2014.01.065

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, (50µm i.d.; effective length, 40cm) was used at 25°C; the applied voltage was 20kV. The background electrolyte solution consisted of 50mmolL−1 sodium dihydrogen phosphate solution at pH 3.0. Injections were performed using a pressure mode at 50mbar for 45s, with detection by photodiode array detector set at 196nm. 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 1.0–300µgmL−1 (r2=0.9992) and the limit of detection (LOD) and limit of quantitation (LOQ) were 0.2µgmL−1 and 1.0µgmL−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 (p0.05). 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. Representative CZE electropherograms showing peak 1=rhIL-11; peak 2=internal standard (IS); peak 3, 4, 5=degraded forms; peak 6=hydrogen peroxide; peak 7=glycine. (a) BRS-rhIL-11; (b) sample of biopharmaceutical formulation. BRS-rhIL-11 following degradation under conditions (c) basic hydrolysis, (d) acid hydrolysis, (e) photolytic, and (f) oxidative. [Display omitted] •Stability-indicating capillary zone electrophoresis method for rhIL-11.•Correlation between physicochemical methods and bioassay for rhIL-11.•Content/potency assessment of rhIL-11 in biotechnology-derived product.