重组人脑利钠肽活性参考品制备及动脉条法生物学活性标定

Establishment of reference material for recombinant human brain natriuretic and peptide bioassay

目的:建立重组人脑利钠肽(rhBNP)活性参考品,用于rhBNP生物学活性测定。方法:优化冻干配方,分装冻干rhBNP原液并检测分装精度,拉丝封口后,按中国药典2010年版三部方法,检测无菌检查、pH、水分、外观。用兔动脉条法由2个实验室协作标定其生物学活性。用热加速法对该批rhBNP参考品的稳定性做了初步评价。结果:分装冻干,拉丝封口等工艺制得rhBNP参考品458支,经测定分装精度小于1%,水分为0.6%,生物学活性经2个实验室共计10次标定,均值为107 RU.支-1,95%的可信范围为97～118 RU.支-1,经热动力学分析,该rhBNP参考品活性降低1%,需要10年以上的时间,说明稳定性较好,其他检测项目均符合标准品一般制备要求。结论:冻干人脑利钠肽活性参考品符合中国药典关于标准品制备要求,可用于rhBNP生物学活性测定。

Objective ：To establish the reference material of recombinant human brain natriuretic peptide （rhBNP） for the determination of bioactivity. Method：First, the bulk formulation and the lyophilization were optimized. Then the bulk was filled into ampouls and the precise of the filling was analyzed. Next, the ampouls with liquid were fro- zen dry and sealed by fire. Finally, the test parameters, including the sterility test, pH, moisture content, appearance, were detected. Specially, the potency was measured by hare artery band test in two cooperation labs respectively. The stability of the frozen was estimated by the thermal accelerating test. Results：A total of 458 vials of rhBNP had been obtained and its precise of filling was less than 1%. The moisture contest was 0.6%. The mean potency of bioactivi- ty was 107 RU · vial- 1, and its 95 % credible range was from 97 RU · vial- 1 to 118 RU · vial-1 Thermal accelera- tion test results showed that it would take more than 10 years for 1% of potency reduction. Moreover, the rest param- eters met the preparation requirements. Conclusion：The rhBNP reference material prepared met the general refer- ence material requirement,which could be used for the rhBNP bioactivity determination.