摘要
采用高效液相分子排阻色谱法(HPLC-SEC)分析不同温度、振荡时间和pH条件下缓冲液中重组人复合α干扰素(cIFN)单体降解和聚合程度,以研究cIFN的体外稳定性。分析表明,振荡会引起cIFN同时发生严重的降解和聚合,在振荡15 min后,聚合体质量分数23.8%,降解产物质量分数29.6%。温度和pH会引发严重降解和部分聚合,40°C下放置66 h会产生质量分数16.3%的聚合体和质量分数49.3%的降解产物;在pH 9放置66 h后会产生质量分数11.5%聚合体和质量分数37.7%的降解产物。研究显示HPLC-SEC方法能对不同环境条件下cIFN单体所发生的聚合和降解变化进行精确定量分析。不同环境下cIFN单体容易产生聚合体和降解产物表明:cIFN稳定性较差,蛋白质单体之间作用形式容易受到环境因素影响而发生改变。
The stability of recombinant human consensus interferon-α(CIFN) was studied with high performance liquid size exclusion chromatography on conditions of different temperature, shake and pH. Shake could induce serious cIFN degradation and aggregation, 23.8% aggregates and 29.6% degradation fragments were produced after 15 min shake. The temperature and pH could induce serious degradation and part aggregation, 16.3% aggregates and 49.3% degradation fragments were produced after 66h at 40℃, and 11.5% aggregates and 37.7% degradation fragments were produced after 66 at prig. The result demonstrated that HPLC-SEC can be used to quantitatively determine cIFN aggregates and degradation fragments on different conditions, and that environment change can influence the stability of cIFN easily.
出处
《江南大学学报(自然科学版)》
CAS
2011年第6期721-725,共5页
Joural of Jiangnan University (Natural Science Edition)
基金
国家973计划项目(2007CB714306)
关键词
高效液相分子排阻色谱
人复合α干扰素
聚合体
降解产物
high performance liquid size exclusion chromatography, recombinant human consensus interferon-α, aggregates, degradation product