刘 红 程永刚 潘红春 徐 波 彭 力 杨红涛 郭 伟. 单链聚乙二醇化重组人干扰素ω的制备及其性质J. 药学学报, 2012,47(3): 393-398.
引用本文: 刘 红 程永刚 潘红春 徐 波 彭 力 杨红涛 郭 伟. 单链聚乙二醇化重组人干扰素ω的制备及其性质J. 药学学报, 2012,47(3): 393-398.
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LIU Hong, Cheng Yong-Gang, Pan Gong-Chun, Xu Bo, Pang Li, Yang Gong-Tao, Guo Wei. Preparation and characterization of mono PEGylated recombinant human interferon omegaJ. 药学学报, 2012,47(3): 393-398.
Citation: LIU Hong, Cheng Yong-Gang, Pan Gong-Chun, Xu Bo, Pang Li, Yang Gong-Tao, Guo Wei. Preparation and characterization of mono PEGylated recombinant human interferon omegaJ. 药学学报, 2012,47(3): 393-398.
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单链聚乙二醇化重组人干扰素ω的制备及其性质

Preparation and characterization of mono PEGylated recombinant human interferon omega

    摘要
  • 摘要:

    采用直链PEG-琥珀酰亚胺琥珀酸酯 (mPEG-SS) 选择性修饰重组人干扰素ω (rhIFNω), 离子交换 色谱和凝胶过滤色谱组合分离纯化单链PEG修饰产物 (PEG-rhIFNω), 基质辅助激光解吸附飞行时间质谱(MALDI-TOF MS) 测定单链PEG-rhIFNω的相对分子量, 并利用RP-HPLC和SDS-PAGE对修饰产物进行分析。在优化的工艺条件下, 分离纯化收集液的单链PEG-rhIFNω, 平均含量达182 μg·mL−1, 分离纯化收率超过22%, 纯度大于98%, SDS-PAGE法测得表观分子质量为60 810, MALDI-TOF MS法测得相对分子质量为43 790; 单链PEG-rhIFNω具有典型PEG修饰蛋白的特性, 抗病毒活性保留率为15.0%, 抗原性降低了64倍, 酸稳定性、抗 胰酶水解能力、血清稳定性和热稳定性均显著提高。单链PEG-rhIFNω的药学性质获得显著改善, 有望开发为 安全、长效的新型干扰素。

     

    Abstract:

    The amino group PEGylation of rhIFNω with monomethoxy polyethylene glycol succinimidyl succinate (mPEG-SS, 20 000) was investigated, and the modified mixture was separated and purified by ion exchange chromatography and gel filtration chromatography.  Under the optimized purification conditions, the average content of mono PEG-rhIFNω in the collect liquid reached 182 μg·mL−1.  The average purified yield of mono PEG-rhIFNω exceed to 22%, and the purity of mono PEG-rhIFNω was greater than 98% by SDS-PAGE and RP-HPLC.  Relative molecular mass of mono PEG-rhIFNω was 43 790 detected by MALDI-TOF MS.  The apparent molecular mass measured by SDS-PAGE was about 60 810.  The purified PEG-rhIFNω has the characteristics of typical PEGylated protein.  Activity reservation rate of mono PEG-rhIFNω was 15.0%, while the antigenicity decreased by at least 64 folds.  In addition, the acid stability, thermal stability and stability in serum and trypsin solution of mono PEG-rhIFNω were markedly better than those of the rhIFNω.  The pharmacological properties of mono PEG-rhIFNω were significantly improved.  The prepared PEG-rhIFNω might be developed to a novel safe and long-acting interferon.

     

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