检测恒河猴血清中trastuzumab浓度的一种直接酶联免疫竞争法(英)

A direct competitive ELISA for determination of trastuzumab concentration in rhesus monkey serum

采用ELISA法建立检测恒河猴血清中trastuzumab的酶联免疫竞争法,为研究人体内trastuzumab的药物动力学学和药效学提供依据。方法的测量范围是1～100μg/mL,最低检测限为1.0μg/mL。板内精密度范围91%～107%,相对标准偏差为1.5%～4.9%。板间精密度范围102%～110%,相对标准偏差为2.7%～15.4%。方法中未显示与重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白、重组抗CD20单克隆抗体、丙种球蛋白等的交叉反应。此方法的特异性、灵敏度、精密度和准确度均满足恒河猴血清样品的分析,是检测猴和人体内trastuzumab的理想方法。

A competitive enzyme-linked immunosorbent assay has been developed for measuring trastuzumab in rhesus monkeys serum.Specifically,the assay was developed to measure trastuzumab in human serum to facilitate future investi-gations of trastuzumab's pharmacokinetics and pharmacodynamics in the human.ELISA working range of this assay was1-100μg/mL,corresponding to a limit of quantification of1.0μg/mL in rhesus monkey serum.Intra-assay recoveries of trastuzumab in monkey serum ranged from91%to107%with RSD values ranging from1.5%to4.9%.Inter-assay recov-eries of trastuzumab in monkey serum ranged from102%to110%with RSD values ranging from2.7%to15.4%.The as-say has no appreciable cross reactivity with RhII TNFR-antibody fusion protein,recombinant chimeric mouse human MAb to CD20and gamma globulin,making this an ideal assay to determine plasma pharmacokinetics of trastuzumab in monkey and human.The assay has sufficient sensitivity,precision,accuracy and specificity for the analysis of rhesus monkey serum samples.