HL60-pNL3.2报告基因法在注射用脑蛋白水解物热原检测中的应用研究  

Application of HL60-pNL3.2 Reporter Gene Assay to Pyrogen Testing of Brain Protein Hydrolysate for Injection

作　　者：王灿 刘荔桢 郑璐侠 陈钢 邵泓 Wang Can;Liu Lizhen;Zheng Luxia;Chen Gang;Shao Hong(NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai Institute for Food and Drug Control,Shanghai 201203,China)

机构地区：[1]上海市食品药品检验研究院、国家药品监督管理局治疗类单抗质量控制重点实验室,上海201203

出　　处：《中国药师》2022年第11期2029-2032,共4页China Pharmacist

基　　金：上海市科委“科技创新行动计划”仪器共享配套操作与应用技术研究课题(编号:21142202900);上海市研发公共服务平台项目(编号:19DZ2294600)。

摘　　要：目的:探讨HL60-pNL3.2报告基因法在注射用脑蛋白水解物热原检测中的应用。方法:参照2020年版中国药典细菌内毒素检查法(通则1143),确定注射用脑蛋白水解物最小有效浓度。参照中国药典《体外热原检查法(报告基因法)》公示稿,干扰实验探讨脑蛋白水解物对热原测定是否存在干扰,并用本实验室建立的HL60-pNL3.2报告基因法对8个厂家生产的16批脑蛋白水解物进行热原测定。结果:注射用脑蛋白水解物最小有效稀释浓度为0.25 mg·ml^(-1)。1 mg·ml^(-1)浓度的脑蛋白水解物溶液对热原测定无干扰。用建立的方法对8个厂家生产的16批注射用脑蛋白水解物进行热原测定,结果均符合规定。结论:HL60-pNL3.2报告基因法具有不使用动物、操作简单、快速、检测热原谱广,可对热原进行定量的优点,可用于注射用脑蛋白水解物热原检测。Objective: To explore the application of HL60-pNL3.2 reporter gene assay to pyrogen detection of brain protein hydrolysate for injection. Methods: According to the general rule 1143 in Chinese Pharmacopoeia(2020 edition), the minimum effective concentration of cerebroprotein hydrolysate for injection was calculated. According to the publication draft named as “In vitro pyrogen test(reporter gene assay)” in Chinese Pharmacopoeia, the interference test was conducted to explore whether the brain protein hydrolysate solution interferes with the pyrogen determination. According to the requirements of publication draft, HL60-pNL3.2 reporter gene assay established in our laboratory was used to detect the pyrogen in 16 batches of brain protein hydrolysate for injection produced by 8 manufacturers.Results: The minimum effective concentration of cerebroprotein hydrolysate for injection was 0.25 mg·ml^(-1). And there was no interference to the pyrogen determination on the concentration of 1 mg·ml^(-1). The pyrogen in 16 batches of brain protein hydrolysate for injection from 8 manufacturers was determined by HL60-pNL3.2 reporter gene assay, and the pyrogen detection results all met the requirements.Conclusion: HL60-pNL3.2 reporter gene assay has the advantages of not using animal, simple operation, fast, wide pyrogen spectrum and pyrogen quantification, which can be used to detect pyrogen in brain protein hydrolysate for injection.

关 键 词：热原测定法 报告基因法 脑蛋白水解物 

分 类 号：R927.12[医药卫生—药学]