间接ELISA法测定猕猴血清中重组人源化抗EGFR单克隆抗体的含量  

Indirect ELISA Determination of Recombinant Human Anti-EGFR Monoclonal Antibody in Rhesus Serum

作　　者：于景敏[1,2] 朱晓霞 孟志云 窦桂芳 刘江林[1,3] 冀希炜 吴卓娜 胡萍[4] 盖文琳[4] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850 [2]北华大学药学院,吉林吉林132000 [3]沈阳药科大学,辽宁沈阳110016 [4]中国医学科学院细胞工程研发中心,北京100851 [5]不详

出　　处：《解放军药学学报》2009年第4期308-311,共4页Pharmaceutical Journal of Chinese People's Liberation Army

基　　金：重大新药创制;No.2009ZX09102

摘　　要：目的建立猕猴血清中重组人源化抗EGFR单克隆抗体的测定方法,为药代动力学研究提供简单快速的方法。方法采用EGFR包外区包被酶标板、HRP标记的IgG-Fc段为标记抗体,建立定量检测抗EGFR单克隆抗体的间接ELISA法,并对其特异性、精密度、准确度、稳定性和稀释效应进行确证。结果在0.46～14.56ng·mL-1浓度范围内,测定方法具有较好的logistic曲线拟合关系,最低检测限为0.46ng·mL-1,组内及组间精密度的RSD分别为6.10%～9.81%,10.17%～11.93%。结论该方法简便、准确、特异性强,精密度及准确度均符合药代动力学要求,可用于猕猴及人血清中抗EGFR单克隆抗体的检测。Aim To establish a method for determination of recombinant human anti-epidermal growth factor receptor（EGFR）monoclonal antibody in rhesus serum and provide a new simple method for studies of pharmacokinetics.Methods The microplate was coated with an extracellular domain of EGFR.HRP labeled IgG-Fc was used as the labelled antibody.The established ELISA was evaluated in terms of specificity,precision,accuracy,stability and dilution effect.Results A good logistic curve relationship was obtained between the OD of anti-EGFR monoclonal antibody and the concentration of anti-EGFR monoclonal antibody over the range of 0.46ng·mL-1 to 14.56ng·mL-1.The lower limit of quantitation was 0.46ng·mL-1.Intra-assay and inter-assay variability were RSD 6.10%～9.81% and 10.17%～11.93%,respectively.Conclusion The method is simple,specific and accurate.It can be employed to the study of pharmacokinetics in rhesus serum samples.

关 键 词：重组人源化抗EGFR单克隆抗体 间接ELISA法 猕猴血清 

分 类 号：R927.2[医药卫生—药学]