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3D-FACS-ANALYSIS-ADCC NK-cells

机译:3D-FACS-ANALYSIS-ADCC NK细胞

摘要

1. A method for evaluating the in vitro effector function of an antibody, which consists in incubating a three-dimensional spheroid or aggregate that contains tumor cells and natural killer cells with an antibody. The method according to claim 1, which consists in the following stages, in which: a) natural killer cells and tumor cells are mixed, b) approximately 10 cells per 200 μl are added to the wells of the multi-well plate, c) the multi-well plate is centrifuged and thereby inducing the formation of a three-dimensional spheroid or aggregate, d) add the antibody to the wells of the multi-well plate, e) incubate the multi-well plate for a period of about 20 to about 72 hours, e) analyze cells in the wells of the multi-well plate while using a cell sorter, and thereby the fluorescence excitation evaluated antitela.3 effector function. The method according to claim 2, characterized in that the natural killer cells are human natural killer cells. The method according to one of the preceding paragraphs, characterized in that the natural killer cells and tumor cells are mixed in a ratio of 10: 1 to 1: 10.5. The method according to claim 4, characterized in that the ratio is from 1: 2 to 1: 4.6. The method according to claim 2, characterized in that the incubation period is from about 20 to about 28 hours. The method according to claim 2 or 6, characterized in that the centrifugation is carried out at 1000 rpm for 10 minutes The method according to claim 2 or 6, characterized in that the tumor cell is a lymphoma cell. The method of claim 8, wherein the lymphoma cell is a Raji cell or SU-DHL4 cell, or Z138 cell. The method according to claim 2
机译:1.一种评估抗体的体外效应子功能的方法,该方法包括将含有肿瘤细胞和天然杀伤细胞的三维球体或聚集体与抗体一起孵育。 2.根据权利要求1所述的方法,其包括以下步骤,其中:a)将自然杀伤细胞和肿瘤细胞混合,b)将每200μl大约10个细胞加入到多孔板的孔中,c)离心多孔板,从而诱导三维球体或聚集体的形成,d)将抗体添加到多孔板的孔中,e)将多孔板孵育约20分钟约72小时,e)使用细胞分选仪分析多孔板孔中的细胞,从而通过荧光激发评估抗tela.3效应子功能。 3.根据权利要求2所述的方法,其特征在于,所述天然杀伤细胞是人天然杀伤细胞。根据前述段落之一的方法,其特征在于,天然杀伤细胞和肿瘤细胞以10∶1至1∶10.5的比例混合。 5.根据权利要求4所述的方法,其特征在于,所述比率为1∶2至1∶4.6。 3.根据权利要求2所述的方法,其特征在于,所述孵育时间为约20至约28小时。 7.根据权利要求2或6所述的方法,其特征在于,所述离心以1000rpm进行10分钟。7.根据权利要求2或6所述的方法,其特征在于,所述肿瘤细胞为淋巴瘤细胞。 9.根据权利要求8所述的方法,其中所述淋巴瘤细胞是Raji细胞或SU-DHL4细胞或Z138细胞。 3.根据权利要求2所述的方法

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