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首页> 外文期刊>Separation and Purification Technology >Phytohemagglutinin isolectins extracted and purified from red kidney beans and its cytotoxicity on human H9 lymphoma cell line
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Phytohemagglutinin isolectins extracted and purified from red kidney beans and its cytotoxicity on human H9 lymphoma cell line

机译：红芸豆中提取的植物血凝素异凝集素及其对人H9淋巴瘤细胞系的细胞毒性

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摘要

Five isolectins, L4, L3E1, L2E2, U E3 and E4, were purified from red kidney bean (Phaseolus vulgaris) by affinity chromatography on a porcine thyroglobulin bound Sepharose~R 4B column, and subsequent separation by Sulfopropyl Sephadex C-50 ion exchange chromatography. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), one clear band appeared for each isolectin, which confirmed the high purity. The decreased haemagglutination activity in an order of E4, L1 E3, L2 E2, L3 E1and L4 further proved the high separation efficiency of the two columns. Low concentration (1 μg/mL) of E4 was able to stimulate the proliferation of human H9 lymphoma cells but strong cytotoxicity was observed at higher concentration. The H9 cell viability was inhibited by the five isolectins in a dose-dependent manner. The concentrations that inhibited 50% of the H9 cell viability were determined to be 3.385 μM for L4, 7.778 μM for L3 E1, 9.939 μM for L2 E2,20.840 μM for L1 E3 and 33.004 μM for E4, respectively. L4 showed more potent cytotoxicity than E4, which was probably due to stronger binding of L4 with the cell-surface carbohydrates than E4. It may also be due to the partial masking of E4 cytotoxicity by the glycoprotein supplemented in the medium.
机译：通过猪甲状腺球蛋白结合的Sepharose〜R 4B柱上的亲和色谱法从红芸豆（菜豆）中纯化了五个异构素L4，L3E1，L2E2，U E3和E4，随后通过磺丙基Sephadex C-50离子交换色谱分离。使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE），每种异凝集素均出现一个清晰的条带，从而证实了其高纯度。血凝活性降低的顺序依次为E4，L1 E3，L2 E2，L3 E1和L4，进一步证明了这两种色谱柱的分离效率很高。低浓度的E4（1μg/ mL）能够刺激人H9淋巴瘤细胞的增殖，但在较高的浓度下却观察到强的细胞毒性。五个异凝集素以剂量依赖的方式抑制H9细胞的活力。抑制H9细胞活力50％的浓度经测定分别为L4为3.385μM，L3 E1为7.778μM，L2 E2为9.939μM，L1 E3为20.840μM和E4为33.004μM。 L4具有比E4更强的细胞毒性，这可能是由于L4与细胞表面碳水化合物的结合比E4更强。这也可能是由于培养基中补充的糖蛋白部分掩盖了E4细胞毒性。

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