摘要
建立了一种HTRF检测方法，可以快速、简便、稳定地测定重组单克隆抗体的含量。将检测试剂、标准品及待测样品加入96孔板，孵育后即可在酶标仪上读取荧光信号值，用软件对信号值进行四参数曲线拟合，绘制标准曲线，根据标准品四参数方程计算待测样品中抗体的含量；同时对方法进行了验证。结果表明：该方法专属性强，抗体含量测定范围为25～1000ng/ml，线性相关系数在0．99以上，回收率范围为92．8％～103．6％，平均为98．9％；该法精密度较好，精密度的变异系数范围为0．2％～11．6％，平均为4．3％。适用于生物制药工艺开发过程中细胞培养上清、纯化产物及制剂中抗体含量的测定。
A rapid and simple antibody analysis method based on HTRF technology was established to measure concentration of recombinant monoclonal antibody in both cell culture medium and purified antibody solution. APC-labeled antigen,antibody samples or controls, and Eu-labeled anti-human IgG are added into 96 well plate （Black） , incubate for some time, followed by fluorescence reading with microplate reader. Standard curve was generated by 4-PL model, antibody concentration in tested samples was calculated accordingly by software. The method qualification results showed that the method is antigen-antibody specific, with accuracy close to 98.9% , precision close to 4.3% , and cell line screening, bio-process monitoring, and linearity range 25 - 1000 ng/ml. This method can be used for quality control during antibody drug development.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2013年第12期64-68,共5页
China Biotechnology
