Determination of recombinant Interferon-α2 in E. coli periplasmic extracts by reversed-phase high-performance liquid chromatography
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Journal of Chromatography B
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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.
Como referenciar
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. Determination of recombinant Interferon-α2 in E. coli periplasmic extracts by reversed-phase high-performance liquid chromatography. Journal of Chromatography B, v. 1072, p. 193-198, 2018. DOI: 10.1016/j.jchromb.2017.11.023. Disponível em: http://repositorio.ipen.br/handle/123456789/28449. Acesso em: 26 Mar 2026.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.