Evaluation of recombinant human interferon beta 1b by liquid chromatography methods and bioassay

Abstract

Recombinant human interferon beta 1b (rhIFNβ−1b) is clinically used to treat multiple sclerosis. A reversed−phase liquid chromatography (RP−LC) method was carried out on a Jupiter C 4 column (250 mm × 4.6 mm i.d.). The mobile phase A consisted of 0.1% trifluoroacetic acid (TFA) in water, and the mobile phase B was acetonitrile with 0.1% TFA run at a flow rate of 1.0 mL/min. A size exclusion liquid chromatography (SE−LC) method was carried out on a BioSep−SEC−S 2000 column (300 mm × 7.8 mm i.d.). The mobile phase consisted of 1 mM monobasic potassium phosphate, 8 mM sodium phosphate dibasic and 200 mM sodium chloride buffer pH 7.4, run isocratically at a flow rate of 0.8 mL/min. Retention times were 31.87 and 17.78 min, and calibration curves were linear over the concentration range of 1−200 µg/mL (r 2 = 0.9998) and 0.50−200 µg/mL (r 2 = 0.9999), respectively, for RP−LC and SE−LC, with detection at 214 nm. Liquid chromatography (LC) methods were validated and employed in conjunction with the in vitro bioassay to assess the content/potency of rhIFNβ−1b, contributing to improve the quality control and to ensure the efficacy of the biotherapeutic.

Alvos Moleculares dos Fármacos no Tratamento da COVID-19

“…Os interferons betas representam a primeira classe de terapias modificadoras de doenças (DMTs), contribuindo consideravelmente para a compreensão dos mecanismos imunomoduladores. O interferon beta humano (hIFN-β) é uma glicoproteína hidrofóbica que contém 166 aminoácidos produzidos por fibroblastos (SILVA et al, 2019). O IFNβ-1b liga-se aos receptores de interferon tipo I, e, aproximadamente, 100 proteínas imunomoduladoras e antivirais são ativadas por sua fosforilação e dimerização (SIRBU et al, 2020).…”