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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,pH8.0)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 study the isolation and purification of crude protein from cobra venom. METHODS: Crude pure Naja (Naja naja) venom was isolated by ion exchange chromatography. The fractions were characterized by high performance liquid chromatography (HPLC) assay, content determination and anti-tumor activity test. RESULTS: Tris-HCl buffer solution (20 mmol·L-1, pH 8.0) was used as the elution system. The crude extract of cobra venom was separated on a DEAE-sephacel column with 0-0.5 mol·L-1 Sodium chloride Tris-HCL solution was subjected to a linear gradient elution, and 4 elution peaks were obtained. The first peak eluted peak anticancer activity contains two types of hydrophobic, UV absorption characteristics of different protein mixtures; the other three peak fractions corresponding to a single fraction. Conclusion: Coleus venom crude fractions can inhibit the growth of cancer cell lines in vitro.