Duensing Thomas D, Watson Susan R
Cold Spring Harb Protoc. 2018 Feb 1;2018(2):2018/2/pdb.prot093815. doi: 10.1101/pdb.prot093815.
A useful feature of therapeutic antibodies is the ability to kill the cells to which they bind. Antibodies are capable of mediating cell killing in a variety of ways. Apoptosis, complement-mediated mechanisms, and antibody-dependent cellular cytotoxicity (ADCC) are all effects that can be assayed to characterize lead antibody candidates. Extensive, multidose characterizations of a series of candidates can be performed in a short amount of time using assays developed for high-throughput flow cytometry systems. Antibodies that contain the Fc portion of the human IgG1 can activate complement-mediated killing. In the ADCC method described here, cytotoxicity is mediated mostly by natural killer (NK) cells. Thus, if an antibody binds to its target on the surface of a tumor cell, Fc receptors on the surface of the NK cells (effector cells) recognize the bound antibody. This leads to the release of cytotoxic granules containing perforin, granzymes, and interferon γ, a cytokine that can stimulate other cells of the immune system such as T cells.
治疗性抗体的一个有用特性是能够杀死它们所结合的细胞。抗体能够通过多种方式介导细胞杀伤。凋亡、补体介导的机制以及抗体依赖性细胞毒性（ADCC）都是可以用来表征潜在抗体候选物的效应。使用为高通量流式细胞术系统开发的检测方法，可以在短时间内对一系列候选物进行广泛的多剂量表征。含有人类IgG1的Fc部分的抗体可以激活补体介导的杀伤作用。在这里描述的ADCC方法中，细胞毒性主要由自然杀伤（NK）细胞介导。因此，如果抗体与肿瘤细胞表面的靶标结合，NK细胞（效应细胞）表面的Fc受体就会识别结合的抗体。这会导致含有穿孔素、颗粒酶和干扰素γ的细胞毒性颗粒释放，干扰素γ是一种可以刺激免疫系统其他细胞（如T细胞）的细胞因子。
