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白细胞介素-11(IL-11)为血小板生长因子,可直接刺激造血干细胞和巨核细胞母细胞的增殖.并诱导巨核细胞的成熟、从而增加血小板的生成.1997年11月由美国Genetic Instirute研制的rhIL-11通过FDA审评.在国内一度成为新的开发热点.在对成都地奥九泓制药厂生产的大肠杆菌表达的重组人白细胞介素-11(rhIL-11)的检定过程中,参照《重组DNA质量控制要点》,研究国内外标淮,结合制品特点,建立了rhIL-11质量标准,对我国第一家送检的样品进行了质量检定.rhIL-11的为179个氨基酸,比天然成熟的人IL-11在N端多一个氨基酸,分子量为19kD,等电点为11.7.虽然厂家基本能按《重组DNA质量控制要点》的要求制定质量标准,但由于缺少有效的HPLC分析方法,肽图分析往往不够准确;活性测定缺少国际标准品,比活性无法确定.针对这些问题我们进行了以下4个方面的研究.
Interleukin-11 (IL-11) is a platelet-derived growth factor that directly stimulates the proliferation of hematopoietic stem cells and megakaryocyte blasts and induces the maturation of megakaryocytes, thereby increasing the production of platelets. In November 1997, it was developed by Genetic Instirute Of rhIL-11 passed the FDA review and was once a new development hotspot in China.In the process of the assay of recombinant human interleukin-11 (rhIL-11) expressed by E.coli produced by Chengdu Diou Jiuhong Pharmaceutical Factory, With reference to the “Essentials of Quality Control of Recombinant DNA”, the quality standards of rhIL-11 were established based on the domestic and international standards and the characteristics of rhIL-11, and the quality of rhIL-11 samples was tested.The samples of rhIL-11 were 179 amino acids, Compared with the natural mature human IL-11, there is one more amino acid at the N-terminus, the molecular weight is 19kD, and the isoelectric point is 11.7. Although the manufacturer can basically set the quality standard according to the requirements of “Essentials of Recombinant DNA Quality Control”, due to lack of effective HPLC analysis Methods, peptide mapping analysis is often not accurate enough; activity determination of the lack of international standards, specific activity can not be determined. In response to these problems we conducted the following four aspects.