论文部分内容阅读
目的 为了经济快速分离眼镜王蛇(Ophiophagushannah,Oh)蛇毒中的毒素成分。 方法 用普通离子交换剂于高效液相色谱柱 (HPLC) TSKgel SP-Toyopearl 65 0 SF (4× 1 5 0 mm)层析法 ,实验取得最佳分离条件后 ,将蛇毒样品上柱后进行梯度洗脱 ,各洗脱峰收集后在 Cosmosil 5 C4-AR-3 0 0柱 (4 .6× 1 5 0 mm)上进行逆相 HPLC分析。非单峰组分再进行 HPLC凝胶过滤柱TSKgel Toyopearl HW-40 Fine(4× 2 5 0 mm)层析 ,层析峰组分再进行 HPLC逆相分析。 结果 眼镜王蛇毒经HPLC离子交换柱层析获得了 1 6个蛋白组分 ,其中有 5个组分经逆相 HPLC分析单一组分 ;另外的复合性组分再进行 HPLC凝胶过滤柱层析后又得到 5个单峰蛋白组分。 结论 HPLC离子交换柱层析对分离蛇毒蛋白很有实用价值 ,特别是蛇毒样品量少的情况下 (1 0 ug)也能较好分离。还具有分离时间短 (1 h左右 ) ,无须低温条件等优点。HPLC凝胶过滤柱层析可进一步使蛋白组分得到提纯
Objective To quickly and economically isolate the toxins from snake venom of Ophiophagushannah (Oh). Methods A common ion exchanger was used for high performance liquid chromatography (HPLC) TSKgel SP-Toyopearl 65 0 SF (4×150 mm) chromatography. After obtaining the best separation conditions, the venom sample was subjected to gradient After elution, each elution peak was collected and subjected to reverse phase HPLC analysis on a Cosmosil 5 C4-AR-30 column (4.6×150 mm). Non-unimodal components were further subjected to HPLC gel filtration column TSKgel Toyopearl HW-40 Fine (4×250 mm) chromatography, and the chromatographic peak components were subjected to HPLC reverse phase analysis. RESULTS: Sixteen protein components were obtained by HPLC ion exchange chromatography on king cobra venom, of which five components were analyzed by reverse phase HPLC. The other components were subjected to HPLC gel filtration chromatography. After that, five unimodal protein components were obtained. Conclusion HPLC ion exchange column chromatography is very useful for the separation of snake venom proteins, especially when the amount of venom sample is small (10 μg). It also has the advantages of short separation time (about 1 h) and no need for low temperature conditions. HPLC gel filtration column chromatography further refines protein fractions