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用紫外扫描光谱、MALDI-TOF MS等电聚焦电泳、毛细管电泳、圆二色光谱、肽图等手段对单链PEGrhIFNω的性能、组分和结构进行表征.结果表明,单链PEG-rhIFNω的最大紫外吸收波长未改变,为280.6nm;等电点由9.7下降到7.35~8.60;质谱分析发现,单链PEG-rhIFNω中仍含有PEG多点修饰组分;圆二色光谱分析发现,单链PEG-rhIFNω二级结构中同时具有典型α螺旋和无规卷曲结构,而rhIFNω没有无规卷曲结构;毛细管电泳分析发现,单链PEG-rhIFNω含4种位置异构体.肽图分析推测,单链PEG-rhIFNω的PEG修饰位点应为Lys33-或Lys167-和Lys46-或Lys52-或Lys152-残基.
The performance, composition and structure of single-stranded PEGrhIFNω were characterized by ultraviolet scanning spectroscopy, MALDI-TOF MS and other electrofocusing electrophoresis, capillary electrophoresis, circular dichroism spectroscopy and peptide mapping.The results showed that the maximum single-chain PEG-rhIFNω UV absorption wavelength was unchanged at 280.6nm; isoelectric point decreased from 9.7 to 7.35 ~ 8.60; mass spectrometry analysis found that single-stranded PEG-rhIFNω still contained more PEG modified components; circular dichroism spectroscopy found that single-stranded PEG -rhIFNω secondary structure at the same time have a typical α-helix and random coil structure, and rhIFNω no random coil structure; capillary electrophoresis analysis found that single-chain PEG-rhIFNω contains four positional isomers. Peptide mapping analysis speculated that single-stranded The PEG modification site for PEG-rhIFN [omega] should be Lys33- or Lys167- and Lys46- or Lys52- or Lys152-residues.