Millipore BioPharma Services, 91 Milton Park, Abingdon, Oxfordshire OX14 4RY, UK.
J Pharm Biomed Anal. 2012 Apr 7;63:23-8. doi: 10.1016/j.jpba.2012.01.029. Epub 2012 Jan 30.
Two different methods have been developed to measure the binding of therapeutic antibodies to the low affinity human Fc receptor FcγRIII (CD16). The first measures binding of antibody to recombinant soluble receptor by surface plasmon resonance and the second uses flow cytometry to measure antibody binding to cells which express the receptor. Both methods have been formatted as parallel line assays and show high levels of accuracy, precision and linearity, making them suitable for comparability, potency and stability assays. They are both readily able to detect structural differences such as glycosylation, which affect Fc receptor binding. The same approaches can be used to measure the binding of any antibody to any Fc receptor. These assays show greater internal precision and long-term reproducibility than traditional cell-based assays such as antibody-dependent cell-mediated cytotoxicity. A combinational approach with a target binding might be appropriate for routine drug batch release as these assays are likely to be significantly more sensitive to small changes in drug structure or activity.
已经开发出两种不同的方法来测量治疗性抗体与低亲和力人 Fc 受体 FcγRIII（CD16）的结合。第一种方法通过表面等离子体共振测量抗体与重组可溶性受体的结合，第二种方法使用流式细胞术测量抗体与表达受体的细胞的结合。这两种方法都已被格式化作为平行线测定法，显示出高度的准确性、精密度和线性度，使其适用于可比性、效力和稳定性测定。它们都能够轻易地检测到影响 Fc 受体结合的结构差异，如糖基化。同样的方法可以用于测量任何抗体与任何 Fc 受体的结合。与抗体依赖性细胞介导的细胞毒性等传统基于细胞的测定法相比，这些测定法具有更高的内部精密度和长期重现性。与靶标结合的组合方法可能适用于常规药物批次放行，因为这些测定法可能对药物结构或活性的微小变化更为敏感。
