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天花粉蛋白(TCS)是一种单链致核糖体失活的蛋白,用细胞融合技术筛选到一株分泌抗该蛋白单抗的杂交瘤细胞株(T1A9),并将单抗T1A9制成免疫亲和凝胶,用此亲和凝胶纯化以TCS和抗人黑色素瘤单抗Ng76构建的免疫毒素,能有效地分离掉样品中的游离抗体,提高免疫毒素的毒效。体外细胞毒性实验表明,亲和纯化的免疫毒素对黑色素瘤细胞(M21)的杀伤作用比分子筛(SephacrylS-200)层析后的样品提高了6.3倍。它对M21细胞的半致死浓度(IC_(50))为1.0×10 ̄(-10)mol/L。比TCS和Ng76的混合物的毒性(IC_(50)=2.5×10 ̄(-7)mol/L)提高了2500倍。TCS是制备免疫毒素的一种有效和常用的“弹头”,用此法纯化这类免疫毒素将会有一定的普遍性。
Trichosanthin (TCS), a single-stranded ribosome-inactivated protein, was screened by a cell fusion technique for a hybridoma cell line (T1A9) secreting the monoclonal antibody and the monoclonal antibody T1A9 And gel, using this affinity gel purified with TCS and anti-human melanoma monoclonal antibody Ng76 constructed immunotoxin, can effectively isolate the free antibodies in the sample to improve the toxic effects of immunotoxins. In vitro cytotoxicity assays showed that the cytotoxic activity of the affinity purified immunotoxin on melanoma cells (M21) was 6.3-fold more than that of the sample after chromatography on Sephacryl S-200. Its half-lethal concentration (IC 50) of M21 cells was 1.0 × 10 -10 mol / L. The toxicity (IC 50 = 2.5 × 10 -7 mol / L) was 2,500 times higher than that of the mixture of TCS and Ng76. TCS is an effective and commonly used “warhead” for the preparation of immunotoxins. Purification of these immunotoxins by this method will be somewhat universal.