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免疫毒素(IT)是目前较为理想的一类 肿瘤导向治疗药物。己引起国内外研究者的重 视,我们将多克隆抗甲胎蛋白抗体(aAFP) 与蓖麻毒素A链(RTA)偶联成免疫毒素aAEP -RTA,并采用荧光染色、流式细胞仪技术测 定该免疫毒素的体外细胞毒效应。 将纯化的山羊抗大鼠AFP抗体与RTA通过 二硫吡啶琥珀酰丙亚胺酸酯(SPDP)法连接, 测得偶联物中的aAFP与RTA的摩尔比为1:1。 用表达AFP抗原的大鼠腹水型肝癌AH66细胞 为靶细胞,以不表这AFP的白血病细胞系P388 为对照细胞。调细胞悬液浓度至2×10~5个 /ml,置离心管中,再按以下分组加药,药量 0.5ml/管:(l)对照:加RPMI1640培养液;
Immunotoxin (IT) is currently an ideal class of tumor-directed therapeutic drugs. Has attracted the attention of researchers at home and abroad, we will polyclonal anti-alpha-fetoprotein antibody (aAFP) and ricin A chain (RTA) conjugated into the immunotoxin aAEP-RTA, and measured by fluorescence staining, flow cytometry The immunotoxin has an in vitro cytotoxic effect. The purified goat anti-rat AFP antibody was ligated with RTA by disulfiratosuccinyl succinimate (SPDP) method and the molar ratio of aAFP to RTA in the conjugate was determined to be 1:1. Aspirin-derived hepatoma AH66 cells expressing AFP antigen were used as target cells, and the AFP leukemia cell line P388 was used as a control cell. Adjust the cell suspension concentration to 2×10~5 cells/ml, place it in a centrifuge tube, and then add the drug in the following groups. The dose is 0.5ml/tube: (1) Control: add RPMI1640 medium;