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目的为了对产品质量进行有效的控制,根据《中国药典》要求,对重组人白介素~(-1)5理化对照品进行分析鉴定。方法参照2010年版《中国药典》三部,测定该样品的生物学活性、蛋白含量、纯度、等电点等指标;通过测定N-末端氨基酸序列、质谱相对分子质量与液质肽图,确证其一级结构。结果该样品的比活性为1.03×107IU·mg~(-1);蛋白含量为(0.879±0.065)mg·m L~(-1);非还原SDS-PAGE纯度为100.0%;分子排阻色谱纯度为100.0%;等电点测定结果为5.2;以上结果均符合质量标准要求。实测质谱相对分子质量(12 900.80)与理论值(12 900.71)一致;氨基酸序列与理论一致,氨基酸覆盖率100%;二硫键连接方式为Cys36-Cys86、Cys43-Cys89,与文献报道相符。结论该理化对照品质量合格、氨基酸序列正确,可用于白介素~(-1)5的常规质量控制。
Objective To control the quality of products effectively, according to the requirements of Chinese Pharmacopoeia, the recombinant human interleukin ~ (-1) 5 physical and chemical reference materials were analyzed and identified. Methods The biological activity, protein content, purity, isoelectric point and other indexes of the samples were determined with reference to the 2010 edition of “Chinese Pharmacopoeia”. By determining the N-terminal amino acid sequence, Primary structure. Results The specific activity of this sample was 1.03 × 107 IU · mg -1. The protein content was (0.879 ± 0.065) mg · m L -1. The purity of non-reducing SDS-PAGE was 100.0%. The molecular weight exclusion chromatography The purity is 100.0%; the isoelectric point is 5.2; the above results are in line with the quality standards. The measured molecular mass (12 900.80) was in agreement with the theoretical value (12 900.71). The amino acid sequence was consistent with the theory and the amino acid coverage was 100%. The disulfide bonding mode was Cys36-Cys86 and Cys43-Cys89, which was in agreement with the literature. Conclusion The physical and chemical quality control standard, amino acid sequence correct, can be used for conventional quality control of interleukin ~ (-1) 5.