摘要
目的:酶联免疫吸附法(ELISA)定量测定食蟹猴血浆中rhCNB多肽的方法学评价。方法:利用双抗体夹心ELISA法,对rhCNB多肽的方法学灵敏度、线性范围、精密度与准确度和回收率进行方法学验证。结果:该方法特异性高,在0.195~12.5 ng/ml的线性范围内,线性关系良好,标准曲线方程为Y=15.1X-0.26,R^2=0.996 8,定量下限(LLOQ)为0.195 ng/ml;该方法的批内精密度CV分别为3.55%、1.39%和4.71%;批间精密度CV分别为1.59%、3.2%和3.8%,均<5%,准确度在91.9%~108.8%之间,方法回收率在88.5%~108.3%之间,<110%,说明该ELISA方法的准确度较高。符合验证要求。结论:双抗体夹心ELISA法灵敏度高、通量大、重复性、准确度及精密度好,可用于生物样品中rhCNB多肽的定量检测。
Objective： To validate an enzyme linked immunosorbent assay（ ELISA） method for the quantification of rhCNB in long-tailed macaque sera. Methods： The linear,sensitivity,accuracy,precision and recovery were determined using ELISA. Results：The present ELISA method had high linearity within 0. 195 ng / ml- 12. 5 ng / ml,the working curve of rhCNB was Y = 15. 1X- 0. 26,R^2= 0. 996 8,the method showed good sensitivity of 0. 195 ng / ml,the accuracy were in the range of 91. 9%- 108. 8%,and the Coefficient of variation（ CV） for inter-assay were 3. 55%,1. 39% and 4. 71%,the intra-assay were 1. 59%,3. 2% and 3. 8%,all less than10%,the recoveries were in the range of 88. 5%- 108. 3%, 〈110%. Thus the method was coincidence with requirement. Conclusion： Double antibody sandwich ELISA assay of rh CNB in long-tailed macaque sera has good sensitivity,accuracy,precision and recovery and it can be used to measure rhCNB concentration in biological samples.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2016年第4期528-531,共4页
Chinese Journal of Immunology
基金
国家自然科学基金资助项目(81160408)
关键词
酶联免疫吸附法
食蟹猴
rhCNB多肽
方法学评价
Enzyme linked immunosorbent assay（ELISA）
Long-tailed macaque
Recombinant human calcineurin B subunit（rhCNB）
Methodological evaluation
作者简介
邵继平（1971年-），女，博士，副教授，主要从事新药安全性评价的研究，E-mail：jpshaosl@163．com．
通讯作者及指导教师：符健（1959年-），男，博士，教授，主任，主要从事新药安全性评价的研究。
