Autor(es): Shytuhina Anastasija, Pristatsky Pavlo, He Jian, Casimiro Danilo R, Schwartz Richard M, Hoang Van M, Ha Sha

Resumo: To effectively support the development of a Chikungunya (CHIKV) virus-like particle (VLP) vaccine, a sensitive - robust high-performance liquid chromatography (HPLC) method that can quantitate CHIKV VLPs - monitor product purity throughout the manufacturing process is needed. We developed a sensitive reversed-phase HPLC (RP-HPLC) method that separates capsid, E1, - E2 proteins in CHIKV VLP vaccine with good resolution. Each protein component was verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) - matrix-assisted laser desorption/ionization time-of-flight (MALDI-ToF) mass spectrometry (MS). The post-translational modifications on the viral glycoproteins E1 - E2 were further identified by intact protein mass measurements with liquid chromatography-mass spectrometry (LC-MS). The RP-HPLC method has a linear range of 0.51-12 ?g protein, an accuracy of 96-106% - a precision of 12% RSD, suitable for vaccine product release testing. In addition, we demonstrated that the RP-HPLC method is useful for characterizing viral glycoprotein post-translational modifications, monitoring product purity during process development - assessing product stability during formulation development.

Palavras-Chave: Analytical characterization; CHIKV; Chikungunya; RP-HPLC; VLP; Vaccine

Imprenta: Journal of Chromatography A, v. 1364, p. 192-197, 2014

Identificador do objeto digital: 10.1016/j.chroma.2014.05.087

Descritores: Chikungunya virus - Biosynthesis ; Chikungunya virus - Cell ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Protein synthesis ; Chikungunya virus - Proteins ; Chikungunya virus - Infectious diseases ; Chikungunya virus - Viral infections ; Chikungunya Virus - Virus ; Chikungunya virus - Vaccine ; Chikungunya virus - Immunology

Data de publicação: 2014

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