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用BlueSepharoseCL-6B凝胶亲和层析法从丝瓜籽中分离纯化了单链致核糖体失活蛋白(ribosomeinac-tivatingprotein,RIP)——lufinB。并将lufinB与抗人黑色素瘤细胞单抗Ng76制成了免疫毒素,命名为lufinB-Ng76,它对体外培养的黑色素瘤细胞M21有很强的抑制作用,IC50为2.5×10-11mol/L,毒性比游离的lufinB提高4000倍,而它对非靶的HeLa细胞的毒性较M21细胞低1200倍。结果提示lufinB用于制备免疫毒素具有良好的应用前景。
The single-stranded ribosome inducing protein (RIP), lufinB, was isolated and purified from the wax gourd by Blue Sepharose CL-6B affinity chromatography. And lufinB and anti-human melanoma monoclonal antibody Ng76 made an immunotoxin, named lufinB-Ng76, it has a strong inhibitory effect on in vitro cultured melanoma cells M21, IC50 is 2.5 × 10-11 mol / L, the toxicity is increased 4000 times compared to free lufinB, and it is 1200 times less toxic to non-target HeLa cells than M21 cells. The results suggest that lufinB has a good application prospect for the preparation of immunotoxins.