adr亚型HBsAg冻干实验参考品的制备与检测  

Preparation and Control Tests on Freeze-dried Laboratory Reference for Quantitative Determination of HBsAg Subtype adr

作　　者：李津 洪云 张国强 王昌华 许毅 张德有 冯素英 汪和睦 赵铠[3] 

机构地区：[1]北京天坛生物制品股份有限公司,北京100024 [2]天津博荟生物技术有限公司检测实验室,天津300192 [3]北京生物制品研究所,北京100024

出　　处：《中国生物制品学杂志》2007年第11期847-850,共4页Chinese Journal of Biologicals

摘　　要：目的制备用于检测重组酵母表达的adr亚型HBsAg含量的实验参考品。方法重组酵母表达的adr亚型HBsAg溶液经纯化后,经3家实验室联合标定蛋白含量,准确稀释并加入保护剂,使抗原最终含量为10μg/ml,分装安瓿后,冷冻干燥熔封,制备成adr亚型实验参考品。再由不同实验室用不同试剂和不同方法,以联合标定的蛋白含量为定量标准,分别验证冻干参考品中adr亚型HBsAg含量及37℃放置不同时间的稳定性。并以adw亚型HBsAg标准作为定量标准,检测adr亚型参考品中HBsAg含量。结果所制备的adr亚型冻干参考品分装准确均匀;以联合标定蛋白含量为标准,不同实验室分别以珠式EIA法和RIA法所测的adr亚型冻干参考品HBsAg含量与目标含量相近,各支参考品间差异小;adr亚型冻干实验参考品37℃放置2～10周后质量稳定。以adw亚型HBsAg参考品为定量标准,所测定adr亚型冻干参考品HBsAg含量,比实际含量约高出0.6倍。结论冻干adr亚型HBsAg实验参考品可用于重组酵母表达adr亚型HBsAg的定量检测。不同亚型HBsAg检测需用各自亚型的参考品作为定量标准。Objective To prepare freeze-dried laboratory reference for the quantitative determination of HBsAg subtype adr expressed in recombinant yeast.Methods The solution of HBsAg subtype adr expressed in recombinant yeast was purified and cooperatively calibrated for protein content by Lowry method by three laboratories,then diluted accurately to a concentration of 10 μ g/ml,added with preservative,filled into ampoules,lyophilized and sealed with fusion to prepare laboratory reference for HBsAg subtype adr.Verify the HBsAg content in the freeze-dried reference and evaluate its stability after storage at 37℃ by different laboratories with defferent reagents,using the cooperatively calibrated protein content as a quantitative standard.The HBsAg content was also determined using the standard for HBsAg subtype adw as a quantitative standard.Results The prepared freeze-dried reference for HBsAg subtype adr was distributed accurately and evenly.Using the cooperatively calibrated protein content as a quantitative standard,all the HBsAg contents in prepared reference verified by bead EIA and RIA were consistent with that expected,and showed no significant difference between containers.After storage at 37℃ for 2～10 weeks,the quality of reference was stable.Using the standard for HBsAg subtype adw as a quantitative standard,the HBsAg content in the prepared reference was about 0.6 times higher than the practical content.Conclusion The prepared laboratory reference HBsAg subtype adr may be used for the quantitative determination of HBsAg subtype adr expressed in recombinant yeast,and the HBsAg of various subtypes shall be determined using reference of the corresponding subtype as a quantitative standard.

关 键 词：ADR亚型 HBSAG 参考品 冷冻干燥 重组酵母 稳定性 

分 类 号：R516.62[医药卫生—内科学] R392-33[医药卫生—临床医学]