重组白介素22-Fc融合蛋白质控方法与质量标准研究

目的: 研究建立重组人白介素22-Fc融合蛋白的质控方法。方法: 通过刺激COLO 205细胞产生白介素10并测定其含量的方法测定该融合蛋白生物学活性;采用SDS-PAGE及高效液相色谱法测定其纯度;采用实时荧光定量PCR方法测定外源DNA残留量;其余项目按中国药典三部(2010年版)要求进行。结果: 该制品3批原液及成品的平均生物学活性分别为参考品的100.6%与103.1%;定量PCR测得3批原液的外源DNA残留量均小于100 pg·剂量^-1;其他各项指标均符合中国药典三部(2010年版)要求。结论: 研究建立的质控方法和质量标准可用于重组人白介素22-Fc融合蛋白的常规质量控制。

Objective: To study and establish the methods and standards for quality control of recombinant interleukin-22-Fc fusion protein. Methods: Biological potency of the fusion protein was evaluated by stimulating COLO 205 cells and determining the content of interleukin-10 produced by stimulation.The purity was analyzed by SDS-PAGE and HPLC.The CHO residual DNA was detected by real-time PCR.Other routine tests were all carried out according to Pharmacopoeia of People's Republic of China (Volume Ⅲ,2010 edition). Results: The average relative biological potency of bulk and products in three batches was 100.6% and 103.1% of reference substances,respectively.The CHO residual DNA of 3 lots of bulk was lower than 100 pg per dose by quantitative PCR.Results of other tests were all in accordance with Pharmacopoeia of People's Republic of China (Volume Ⅲ,2010 edition). Conclusion: The methods and standards which were established could be used for routine quality control of recombinant interleukin-22-Fc fusion protein.

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