目的 研究及探讨四参数模型拟合算法的具体过程,并将该模型应用于细菌内毒素标准品效价标定方法中。方法 将细菌内毒素国家标准品作为标准品,细菌内毒素工作标准品作为待标品。运用HL60-IL6单核细胞激活实验法进行效价标定。以标准品和待标品溶液的反应吸光度值对其相应的浓度进行四参数Logistic拟合,采用非线性最小二乘法进行迭代计算,该拟合算法过程及结果用C语言、softmax软件、R语言以及Microsoft Office Excel软件实现。结果 4种分析方式结果一致,拟合度均大于0.99,相对效价值的精度差异在0.1%之内。结论 4种软件均适用于四参数Logistic模型的拟合及效价的计算,科研分析人员可根据实际需求来选用合适的工具。
Abstract
OBJECTIVE To study the algorithm of four-parameter logistic regression and apply the model in the potency calibration of bacterial endotoxin standard. METHODS The potency of endotoxin reference standard was estimated by a novel monocyte activation test,HL60-IL6 assay.The endotoxin national standard was set as standard, while the endotoxin working standard was set as test. The four-parameter logistic model was fitted by the method of nonlinear least squares when observations of standard or sample solutions were plotted against endotoxin concentration. The algorithm was performed by C Language, the RESULTS of which were compared by those of softmax, R Language as well as Microsoft Office Excel. RESULTS The estimated parameters as well as estimated potency were parallel between the four statistical tools. The goodness-of-fit were all over 0.99.The variance was within 0.1%. CONCLUSION The four-parameter logistic model can be established by the four statistical tools. Researchers can select a suitable tool according to demands.
关键词
四参数Logistic模型 /
迭代 /
HL60-IL6法 /
细菌内毒素 /
效价
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Key words
four-parameter logistic model /
iteration /
HL60/IL6 assay /
endotoxin /
potency
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中图分类号：
R915
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参考文献
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脚注
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基金
药品医疗器械审评审批制度改革专项课题资助(ZG2016-4-05)
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