FERNANDA DOS SANTOS ARTHUSO
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Artigo IPEN-doc 24236 Determination of recombinant Interferon-α2 in E. coli periplasmic extracts by reversed-phase high-performance liquid chromatography2018 - DIAS, PAULO V.S.; ARTHUSO, FERNANDA S.; OLIVEIRA, JOAO E.; SUZUKI, MIRIAM F.; SOUSA, JOSE M.; RIBELA, MARIA T.C.P.; BARTOLINI, PAOLO; SOARES, CARLOS R.J.Reversed-phase high-performance liquid chromatography (RP-HPLC) has been used to analyze Interferon α-2 (IFN-α2) as a pure protein or as a pharmaceutical preparation: a method for analyzing periplasmic IFN-α2 directly in osmotic shock extract has, however, never been reported. This work describes an RP-HPLC methodology for the qualitative and quantitative analysis of human IFN-α2a and IFN-α2b directly in bacterial periplasmic extracts or in purified preparations. The analytical method has been set up and validated for accuracy, precision, linearity, sensitivity and specificity. A recovery test indicated an average bias of ∼1%, intra-day and inter-day quantitative determinations presented relative standard deviations always ≤ 5%, while the working sensitivity was of ∼0.3 μg of IFN-α2 (RSD =5%). The method proved to be suitable for detecting and quantifying also glycosylated and oxidized forms and N-methionylated IFN-α2 molecules, it was, however, not able to distinguish between IFN-α2a and IFN-α2b. This rapid methodology allows the application of RP-HPLC as a powerful tool to monitor the production yield and quality of IFN-α2 in osmotic shock fluids, right after, or even during the fermentation process.Resumo IPEN-doc 22087 Aplicação do bioensaio Ba/F3-LLP na determinação da potência biológica da prolactina humana glicosiladfa (G-hPRL)2007 - ARTHUSO, FERNANDA dos S.; SOARES, CARLOS R.J.Artigo IPEN-doc 18110 Expression, purification, and characterization of authentic mouse prolactin obtained in Escherichia coli periplasmic space2012 - SUZUKI, MIRIAM F.; ARTHUSO, FERNANDA S.; OLIVEIRA, JOAO E.; OLIVEIRA, NELIO A.J.; GOULART, HERBERT R.; CAPONE, MARCOS V.N.; RIBELA, MARIA T.C.P.; BARTOLINI, PAOLO; SOARES, CARLOS R.J.Artigo IPEN-doc 14975 Synthesis, purification and characterization of recombinant glycosylated human prolactin (G-hPRL) secreted by cycloheximide-treated CHO cells2010 - HELLER, S.R.; GOULART, H.R.; ARTHUSO, F.S.; OLIVEIRA, T.L.; BARTOLINI, P.; SOARES, C.R.J.Artigo IPEN-doc 15555 Enhancement of human prolactin synthesis by sodium butyrate addition to serum-free CHO cell culture2010 - GOULART, HERBERT R.; ARTHUSO, FERNANDA dos S.; CAPONE, MARCOS V.N.; OLIVEIRA, TAIS L. de; BARTOLINI, PAOLO; SOARES, CARLOS R.J.Sodium butyrate (NaBu) has been used as a productivity enhancer for the synthesis of recombinant proteins in Chinese hamster ovary (CHO) cells. Thus, the influence of NaBu on the production of recombinant human prolactin (hPRL) from CHO cells was investigated for the first time. CHO cell cultures were submitted to a treatment with different concentrations of NaBu (0.25 to 4 mM). Quantitative and qualitative analyses by reverse-phase high-performance liquid chromatography (RP-HPLC) and Western blot or SDS-PAGE, carried out directly on CHO-conditioned medium, showed that the highest hPRL expression was obtained with 1 mM NaBu. In vitro biological assays based on noble rat lymphoma (Nb2) and mouse pro-B lymphoma (Ba/F3-LLP) cells were carried out on purified hPRL. Its bioactivity in the presence of NaBu was not apparently different from that of the First International Reference Reagent of recombinant hPRL (WHO 97/714). Our results show that NaBu increased the synthesis of recombinant hPRL in CHO cells, apparently without compromising either its structure or function.Artigo IPEN-doc 16350 Lymphocytic prolactin does not contribute to systemic lupus erythematosus hyperprolactinemia2010 - PARAIBA, D.B.; SOARES, C.R.J.; BARTOLINI, P.; ARTHUSO, F.S.; BORBA, E.F.; BONFA, E.; BRONSTEIN, M.D.Artigo IPEN-doc 18184 Laboratory production of human prolactin from CHO cells adapted to serum-free suspension culture2012 - ARTHUSO, FERNANDA S.; BARTOLINI, PAOLO; SOARES, CARLOS R.J.