Cedeño-Arias Mercedes, Sánchez-Ramírez Javier, Blanco-Santana Rancés, Rengifo-Calzado Enrique
Center of Molecular Immunology, 216 St and 15 Ave. Atabey, Playa. POBox 16040, Havana 11600, Cuba.
Sci Pharm. 2011 Jul-Sep;79(3):569-81. doi: 10.3797/scipharm.1104-18. Epub 2011 Jul 3.
An ideal test used to characterize a product must be appropriate for the measurement of product quality, manufacturing consistency, product stability, and comparability studies. Flow cytometry has been successfully applied to the examination of antibodies and receptors on membrane surfaces; however, to date, the analytical validation of cytometry based assays is limited. Here we report on the validation of a flow cytometry-based assay used in the evaluation of nimotuzumab binding to cells over-expressing EGFR on cell surface. The assay was validated by examining, assay robustness, specificity, repeatability and intermediate precision. The assay was highly specific, robust for all studied factors except for cell fixation with 1% paraformaldehyde and met criteria for precision with RSD < 2%. In addition the assay has stability-indicating properties evidenced by the ability to detect changes in mAb degraded samples. Most importantly, the assay demonstrated to be useful for its intended use.
用于表征产品的理想测试必须适用于产品质量、生产一致性、产品稳定性和可比性研究的测量。流式细胞术已成功应用于膜表面抗体和受体的检测；然而，迄今为止，基于细胞术的检测方法的分析验证是有限的。在此，我们报告一种基于流式细胞术的检测方法的验证，该方法用于评估尼妥珠单抗与细胞表面过度表达表皮生长因子受体（EGFR）的细胞的结合情况。通过检测该检测方法的稳健性、特异性、重复性和中间精密度对其进行了验证。该检测方法具有高度特异性，除了用1%多聚甲醛固定细胞外，对所有研究因素均具有稳健性，并且符合精密度标准，相对标准偏差（RSD）<2%。此外，该检测方法具有稳定性指示特性，这通过检测单克隆抗体降解样品中的变化的能力得到证明。最重要的是，该检测方法已证明对其预期用途有用。
