Recombinant human interferon analysis in pharmaceutical formulations

• Autor(a) principal: de Andrade, Sinéa Mendes
• Data de Publicação: 2017
• Outros Autores: da Silva, Manuela, Silva, Filipe Soares Quirino da
• Tipo de documento: Artigo
• Idioma: por; eng
• Título da fonte: Vigilância Sanitária em Debate
• Texto Completo: https://visaemdebate.incqs.fiocruz.br/index.php/visaemdebate/article/view/943
• topic: Interferon-alfa; Proteínas; MALDI-TOF; Dicroísmo Circular; Fluorescência; Alpha Interferon; Protein; MALDI-TOF; Circular Dichroism; Fluorescence

Introduction: Due to the interest in the treatment of hepatitis, the industrial production process of INF-α has been developed and perfected over the last few years. Objective: The present work aimed to develop a protocol to characterize the molecular structure of INF-α2b in pharmaceutical formulations by MALDI-TOF mass spectrometry. Method: Initially, a reversed-phase liquid chromatography method was developed to promote the separation of active and minor constituent INF-α2b and human serum albumin, also present in the pharmaceutical formulations, to obtain samples with protein homogeneity revealed by electrophoresis. Samples were hydrolyzed with trypsin and submitted to MALDI-TOF. In order to analyze the molecular structure, a procedure based on immunoaffinity and gel filtration chromatography was developed. Results: Prepared samples by these methods showed protein homogeneity by SDS-PAGE, and were analyzed by circular dichroism and fluorescence, which showed three – dimensional structure degradation. Conclusions: This work provides important data that support the establishment of a protocol for the analysis of INF-α2b in final product, which could replace the traditional peptide mapping by liquid chromatography, with the advantage of resulting in a larger amount of information about the structure of the biopharmaceutical.