Recombinant human interferon analysis in pharmaceutical formulations
DOI:
https://doi.org/10.22239/2317-269X.00943Keywords:
Alpha Interferon, Protein, MALDI-TOF, Circular Dichroism, FluorescenceAbstract
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.Downloads
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