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Assessment of rhIL-11 by Validated SE-LC Method and its Correlation with RP-LC and CZE Methods

[ Vol. 14 , Issue. 3 ]

Author(s):

Ricardo Bizogne Souto, Rafaela Ferreira Perobelli, Bruna Xavier, Fernanda Pavani Stamm Maldaner, Francielle Santos da Silva, Gabriel Lunardi Remuzzi and Sergio Luiz Dalmora*   Pages 191 - 197 ( 7 )

Abstract:


Background: Recombinant human interleukin-11 (rhIL-11) is a cytokine produced by recombinant DNA technology and used clinically to treat chemotherapy-induced thrombocytopenia. A Size Exclusion Liquid Chromatography (SE-LC) method was validated to assess the content/potency of the rhIL-11.

Methods: A BioSep-SEC-S2000 column (300 mm × 7.8 mm i.d.) maintained at 30°C was used. The mobile phase was composed of 0.02 M 2-(N-morpholino)ethanesulfonic acid and 0.5 M sodium chloride buffer, pH 6.0, run isocratically at a flow rate of 0.9 mL min-1, with detection by Photodiode Array (PDA) detector set at 220 nm.

Results: Chromatographic separation was obtained with a retention time of 8.12 min, and the stabilityindicating capability, and specificity was demonstrated. The method was linear over the concentration range of 1.0–200 µg mL-1 (r2= 0.9996) and the Detection Limit (DL) and Quantitation Limit (QL) were 0.23 µg mL-1 and 1.0 µg mL-1, respectively. The accuracy was 99.81% with bias lower than 0.43%. Moreover, method validation showed acceptable results for precision and robustness.

Conclusion: The method was applied to the assessment of rhIL-11 and Higher Molecular Weight forms (HMW) in biopharmaceutical formulations, and the results of content/potencies were correlated to those of a validated Reversed-Phase Liquid Chromatography (RP-LC) and a Capillary Zone Electrophoresis (CZE) methods, showing non-significant differences (p> 0.05). The proposed method and the correlation studies contribute to monitor stability, improve the quality control and ensure the batch-to-batch consistency of the biotherapeutics.

Keywords:

Recombinant Human Interleukin-11, size exclusion liquid chromatography, reversed-phase liquid chromatography, capillary zone electrophoresis, validation, correlation.

Affiliation:

Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria, Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria, Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria, Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria, Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria, Postgraduate Program in Pharmaceutical Sciences, Federal University of Santa Maria, Santa Maria, Postgraduate Program in Pharmaceutical Sciences, Federal University of Santa Maria, Santa Maria

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