屋尘螨变应原制品活性测定方法的建立  

Bioactivity detection method for Dermatophagoides pteronyssinus products

作　　者：王春娥[1] 石继春[1] 王珊珊[1] 陈琼[1] 李茂光[1] 叶强[1] 李凤祥[1] 

机构地区：[1]中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《药物分析杂志》2013年第5期775-778,共4页Chinese Journal of Pharmaceutical Analysis

基　　金：国家高科技研究发展计划(863计划)"疫苗效果和质量评价新技术研究";课题编号:2012AA02A402;国家科技支撑计划"过敏性疾病体内诊断及治疗制剂技术标准研究";课题编号:2008BAI59B04

摘　　要：目的:建立屋尘螨变应原活性测定方法,用于屋尘螨变应原制品的质量控制。方法:分别对屋尘螨原液、不同企业含氢氧化铝的屋尘螨制品进行试验条件的优化,建立荧光酶联免疫竞争抑制法-UniCAP法进行活性测定。结果:在37℃2 h和4℃过夜条件下,屋尘螨原液及制剂、螨变应原注射液的测定结果均无显著性差异;以0.1 mol.L-1磷酸盐缓冲液、0.1 mol.L-1磷酸盐缓冲液-0.5%牛血清白蛋白、0.9%氯化钠溶液作为稀释液,屋尘螨原液的测定结果无显著性差异;采用1∶1的血清稀释度、0.1 mol.L-1磷酸盐缓冲液、4℃过夜的实验条件,对方法的精密度进行考察,重复性和中间精密度RSD分别为0.43%和1.3%;对14批屋尘螨变应原制剂总活性和游离过敏原活性进行测定,总活性均不低于50%,均值为68%(RSD为1.9%),游离过敏原活性均不高于20%,均值为9%(RSD为40%)。结论:该方法重复性好,快速,可用于原液和成品活性测定。可通过测定总活性和游离过敏原活性,对含氢氧化铝的屋尘螨制品进行质量控制。Objective：To establish a detection method for the bioactivity of Dermatophagoides pteronyssinus allergens, and apply for the quality control of Dermatophagoides pteronyssinus products. Methods ： The assay for differentDermatophagoides pteronyssinus products was optimized using different reaction time, temperatures and buffer solutions. Fluorescent enzyme - linked immunosorbent inhibition assay was established for the in vitro bioactivity. Re-suits ： There was no significant difference between at 37 ℃ for 2 h and at 4 ℃ for overnight for the results of the inhibition rate of bulk of D. P, mites allergens of D. P, and Novo - Helisen - Depot. The results of the bulk of D. Pshowed no significant difference when 0.1 mol·L^-1 phosphate buffer, 0.1 mol·L^-1 phosphate buffer with 0.5% bovine serum albumin, or 0.9% sodium chloride solution was used as diluent respectively. The precision of themethod was inspected under the condition of 1：1 dilution of serum pool, 0.1 mol·L^-1 phosphate buffer, 4 ℃ overnight, and the repeatability and intermediate precision were 0.43% （RSD） and 1.3% （RSD） respectively. Forall the 14 batches of mites allergens of D. P, the total activity was no less than 50% with average 68% （ RSD = 1.9 % ） and supernatant allergen activity was no more than 20% with average 9% （RSD = 40% ）. Conclusion：Themethod is repeatable, rapid and suitable for activity detection of both bulk and final products. The results indicate that the established method for detection of total activity and supernatant allergen activity can be applied to thequality control of aluminium hydroxide absorbed Dermatophagoides pteronyssinus products.

关 键 词：屋尘螨 变应原 活性测定 荧光酶联免疫 质量控制 

分 类 号：R917[医药卫生—药物分析学]