眼镜蛇毒粗纯蛋白的柱层析分离及其各馏分的抗肿瘤活性研究

目的: 眼镜蛇毒粗纯蛋白的分离及其馏分抗癌活性研究。 方法: 应用离子交换层析法分离眼镜蛇毒粗纯蛋白,通过高效液相色谱行为分析、含量测定和抗肿瘤活性试验对各馏分进行了性质研究。 结果: 以Tris-HCl缓冲液(20 mmol·L^-1,pH 8.0)为洗脱体系,在DEAE-sephacel填料离子交换柱上对眼镜蛇毒粗纯蛋白以0~0.5 mol·L^-1氯化钠的Tris-HCL溶液进行线性梯度洗脱,可得到4个洗脱峰。第1个洗脱峰馏分抗癌活性最强,含有两类疏水性、紫外吸收特性各异的蛋白混合物;另3个洗脱峰对应馏分蛋白成分较单一。 结论: 眼镜蛇毒粗纯蛋白分离馏分对体外培养的癌细胞株有一定的抑制作用。

Objective: To separate crude protein and purified protein in cobratoxin and analyze anti-tumor activity of its fractions. Methods: The crude protein and purified protein in cobratoxin was separated by ion exchange chromatography. Qualitative study was made on each fraction by means of analysis by reverse-phase chromatography, content determination and anti-tumor activity test. Results: Four elution peaks were obtained when a linear gradient elution was applied to crude protein and purified protein in cobratoxin by adding sodium chloride (0-0.5 mol·L^-1) to Tris-HCl buffer (20 mmol·L^-1,pH 8.0) on DEAE-sephacel ion-exchange column. Among the four elution peaks, the first one showed the highest anti-tumor activity. It contained two types of proteins, which differed in hydrophobicity and ultraviolet absorption, while the other three elution peaks contained single protein respectively. Conclusion: The fractions from cobratoxin have some inhibitory effects on tumor cell lines cultured in vitro.

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