东北林蛙皮肤分泌物中特定范围内蛋白和多肽凝胶层析分析

Protein and Polypeptide Saphadex Chromatography of Skin Secretion of Rana dybowskii

利用凝胶色谱法对东北林蛙皮肤分泌物中蛋白质及多肽组分分子量的分布进行分析。应用葡聚糖凝胶Sephadex G-100、G-50、G-25,利用已知分子量的牛血清蛋白(Mr 66 000)、卵清蛋白(Mr 45 000)、细胞色素C(Mr 11 700)、胰岛素(Mr 5 700)、胰高血糖素(Mr 3 550)等作为标准品,对流动相、洗脱速度、色谱条件等进行反复试验,用0．5％甲醇洗脱液(含0．14 mol／L NaCl)、0．4 ml／min流速,筛选东北林蛙皮肤分泌物中分子量Mr 4 000左右的蛋白质及多肽,分离获得了大量相应分子量范围的蛋白及多肽组分。该实验方法简便,结果重现性好,适合作为生物工程产品的质控方法。

Proteins and polypeptides of a specific range of molecular weights in skin secretion of Rana dybowskii were analyzed by chromatography of Saphadex G-100, G-50, and G-25; BSA （Mr 66 000 ）, OVA（Mr 45 000 ）, cytochrome C （Mr 11 700）, insulin （Mr 5 700） and glucagon （Mr 3 350） were used as molecular markers, screened the secretion at different flow phases, flow speeds and chromatographic conditions, finally washed with 0.5% methanol solution （in 0.14 mol/L NaCl）, A number of proteins and polypeptides of about Mr 4 000 were obtained, with good repetition.