恒河猴血清非特异性凝集因子消除方法的建立  

Establishment of a Method for Eliminating Nonspecific Agglutination Factors in Rhesus Monkey Serum

作　　者：冉艳洪 颜欣欣 宋慧 戴梦雨 王俐梅 杨威 RAN Yanhong;YAN Xinxin;SONG Hui;DAI Mengyu;WANG Limei;YANG Wei(Guangdong Lewwin Pharmaceutical Research Institute Co.,Ltd.,Guangdong Provincial Key Laboratory of Drug Non-Clinical Evaluation and Research,TCM Non-clinic Evaluation Branch of National Engineering Research Center for Modernization of Traditional Chinese Medicine,Guangdong Engineering Research Center for Innovative Drug Evaluation and Research,Guangzhou 510990,China;Guangzhou Bay Area Institute of Biomedicine,Guangzhou 510990,China)

机构地区：[1]广东莱恩医药研究院有限公司,广东省药物非临床评价研究企业重点实验室,国家中药现代化工程技术研究中心中药非临床评价分中心,广东省创新药物评价与研究工程技术研究中心,广州510990 [2]广州湾区生物医药研究院,广州510990

出　　处：《实验动物科学》2025年第1期23-26,共4页Laboratory Animal Science

基　　金：广东省重点领域研发计划资助(2020B1111030005);广东省重点领域研发计划“新药创制”重点专项(2019B020202002);广州市基础与应用基础研究项目(202102020984)。

摘　　要：目的探索恒河猴血清非特异性凝集因子(NSAs)的消除方法。方法首先分别使用1%鸡红细胞和1%豚鼠红细胞对恒河猴血清进行非特异性凝集筛选,然后将非特异性凝集血清样本与不同浓度(1%、5%、10%、15%和20%)的鸡红细胞等体积混匀,分别置室温吸附处理0.5、1 h和2~8℃吸附处理2、4、6、8和12 h后检测被检血清凝集情况,初步确定恒河猴血清NSAs鸡红细胞吸附消除的条件,并对其进行效果验证,从而最终明确恒河猴血清NSAs鸡红细胞吸附消除的最佳红细胞浓度、吸附时间、吸附温度以及消除稳定性。结果(1)恒河猴血清中存在NSAs,使用1%鸡和1%豚鼠红细胞对恒河猴血清进行非特异性凝集筛选其凝集率均为100%。(2)恒河猴非特异性凝集血清使用不同浓度鸡红细胞在室温下吸附处理0.5 h后NSAs去除率为0%~50%,处理1 h后NSAs去除率均为0%;在2~8℃下吸附处理2、4、6、8和12 h后NSAs去除率分别为20%~54%、40%~100%、90%~100%、80%~100%、90%~100%。(3)60份恒河猴非特异性凝集血清在2~8℃下使用5%鸡红细胞吸附处理4和6 h,NSAs的去除率可达90.00%和83.33%。结论本研究成功建立了一种高效、稳定、可靠的恒河猴血清NSAs鸡红细胞吸附消除方法。Objective This study aims to explore a method for eliminating nonspecific agglutination factors(NSAs)in rhesus monkey serum.Method Firstly,1%chicken red blood cells and 1%guinea pig red blood cells were used to screen NSAs in rhesus monkey serum,respectively.Then,serum samples showing nonspecific agglutination were mixed with different concentrations(1%,5%,10%,15%and 20%)of chicken red blood cells in equal volumes,and the agglutination test was carried out by chicken red blood cells after adsorption treatment at room temperature for 0.5 h,1 h and 2-8℃for 2,4,6,8,12 h,respectively.Ultimately,the conditions of adsorpting and eliminating NSAs in rhesus monkey serum were preliminarily determined and verified,included the best chicken red blood cell concentration,adsorption time,adsorption temperature and elimination stability.Result(1)NSAs were existed in rhesus monkey serum,with a 100%agglutination rate was observed when screened using 1%chicken red blood cells or 1%guinea pig red blood cells.(2)The rhesus monkey nonspecific agglutination serum was adsorpted with different concentrations of chicken erythrocytes.The elimination rate of NSAs range from 0%-50%,0%after treated at room temperature for 0.5 h and1 h,respectively.The elimination rates of NSAs range from 20%-54%,40%-100%,90%-100%,80%-100%,90%-100%after treated at 2-8℃for 2,4,6,8 and 12 h,respectively.(3)The elimination rate of NSAs in rhesus monkey serum could reach at 90.00%and 83.33%when treated with 5%chicken erythrocytes at 2-8℃for 4 h and 6 h,respectively.Conclusion This study successfully established an efficient,stable and reliable method for eliminating NSAs in rhesus monkey serum using chicken red blood cells adsorption treatment.

关 键 词：非特异性凝集因子 消除 红细胞吸附 恒河猴 

分 类 号：Q95-3[生物学—动物学]