ABSTRACT
To establish an HPLC method to determine osimertinib mesylate,Agilent ZORBAX Eclipse Plus C18 column (4.6 mm × 250 mm,5 μm) was used with a mobile phase consisting of methanol-buffer solution (20 mmoL/L NaH2PO4,pH 3.0 adjusted with 85％ H3PO4) (50 ∶ 50) at the flow rate of 1.0 mL/min.The detection wavelength was 210 nm,and the column temperature was kept at 35 ℃.The calibration curve was liner over the range from 50％ to 150％ of determination concentration (0.201 1-0.603 2 mg/mL,r =0.999 9).The limit of quantitation (LOQ) and limit of detection (LOD) were 0.32 μg/mL and 0.08 μg/mL,respectively.The contents of osimertinib mesylate in samples were 100.1％,99.5％ and 99.7％.Good chromatographic separation of osimertinib mesylate and its related substances,including synthetic impurities and degradation products,were obtained.The established HPLC method is specific,accurate,simple and durable,and could be used for the determination of osimertinib mesylate.
ABSTRACT
Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. Coli and purified by nickel-nitrilotriacetic acid (Ni2+-NTA) affinity resin column,finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.