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建立和比较基于RAJI细胞的流式细胞法(Flow cytometry assay,FCA)和细胞放射免疫法(Immuno-radiametric assay,IRA)测定重组抗CD20人源化单克隆抗体(r-anti-CD20zumAb)的浓度。两种方法均利用标记后抗体和被检测抗体竞争性结合RAJI细胞表面的CD20抗原,由标记后抗体的荧光或放射性变化而间接反应检测抗体的浓度。FCA和IRA定量范围分别为0.1～100mg/L和0.04～20mg/L,FCA的日内及日间精密度分别小于4.0%和3.0%,准确度为-1.7%～1.1%,IRA的日内及日间精密度值均小于7.0%,准确度为-8.9%～13.2%。方法学确证研究表明,两种方法均具有良好的特异性、灵敏度、精密度和准确度。血浆样品分析结果显示,两种方法具有良好的一致性,是检测猕猴血浆中人源化单克隆抗体浓度的理想方法。
The concentration of recombinant anti-CD20 humanized monoclonal antibody (r-anti-CD20zumAb) was determined by flow cytometry assay (FCA) and cell radioimmunoassay (IRA) . Both methods use the labeled antibody and the test antibody to competitively bind to the CD20 antigen on the surface of the RAJI cell, and the concentration of the antibody is detected indirectly by the fluorescence or radioactive change of the labeled antibody. The FCA and IRA quantitative ranges were 0.1-100 mg / L and 0.04-20 mg / L, respectively. The intra-and inter-day precision of FCA was less than 4.0% and 3.0%, respectively, with the accuracy of -1.7% -1.1% Inter-precision values were less than 7.0%, accuracy was -8.9% ~ 13.2%. Methodological confirmatory studies have shown that both methods have good specificity, sensitivity, precision and accuracy. Analysis of plasma samples showed good agreement between the two methods and was an ideal method for detecting the concentration of humanized monoclonal antibodies in macaque plasma.