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目的:为开发干扰素α-2b新剂型提供纯化和质控的技术指标。方法:用反相离子对色谱法对干扰素α-2b的粗品进行分离纯化;用分子排阻色谱法测定其蛋白纯度。结果:反相离子对色谱法可使纯度为85％粗品提高到95％以上;分子排阻色谱法可通过峰面积比,快速、客观地确定表达蛋白的百分比,对主峰、次峰的分离度大于1.5。结论:反相离子对HPLC法可作为干扰素α-2b的纯化方法;分子排阻色谱法是鉴定基因工程干扰素的不可缺少的指标之一,它可对提纯工艺起到有效的监控作用。
OBJECTIVE: To provide a technical indicator of purification and quality control for the development of a new dosage form of interferon alfa-2b. Methods: The crude interferon α-2b was isolated and purified by reversed-phase ion-pair chromatography. The protein purity was determined by size exclusion chromatography. Results: Reversed-phase ion chromatography (RP-HPLC) could increase the crude product purity to over 95%, and the molecular weight exclusion chromatography (GC-MS) could quickly and objectively determine the percentage of expressed protein by the peak area ratio. Greater than 1.5. CONCLUSION: RP-HPLC method can be used as a method for the purification of interferon alfa-2b. Molecular exclusion chromatography is an indispensable indicator for the identification of genetically engineered interferon. It can effectively monitor the purification process.