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目的 :建立确定胸腺素α1纯度、分子量和等电点等生化特性和生物学活性检测的方法。方法 :应用高效液相色谱、三羟甲基甘氨酸 -SDS -聚丙烯酰胺凝胶电泳、等电聚焦 -聚丙烯酰胺凝胶电泳和3H -TdR掺入法 ,在致有丝分裂原存在的条件下 ,刺激小鼠脾淋巴细胞分裂增殖的能力。结果 :胸腺素α1的纯度为 98% ,其相对分子质量为 3 15 3 ,等电点为3 93,在 1 5 6～ 12 5 μg·mL-1剂量范围内脾淋巴细胞的增殖率明显提高。结论 :本文所用检测方法能准确地测定胸腺素α1的纯度、相对分子质量和等电点 ,有效地测定其生物学活性 ,适用于胸腺素α1的质量鉴定
OBJECTIVE: To establish a method to determine the biochemical characteristics and biological activity of thymosin α1, such as purity, molecular weight and isoelectric point. METHODS: High performance liquid chromatography (HPLC), trimethylolglycine-SDS-polyacrylamide gel electrophoresis, isoelectric focusing-polyacrylamide gel electrophoresis and 3H-TdR incorporation assay were used to detect the presence of mitogen. Stimulation of splenic lymphocyte proliferation in mice. Results: The purity of thymosin α1 was 98%, the relative molecular mass was 3 15 3 and the isoelectric point was 3 93. The proliferation rate of splenic lymphocytes was significantly increased in the dose range of 165-6125 μg · mL-1 . Conclusion: The detection method used in this paper can accurately determine the purity, relative molecular mass and isoelectric point of thymosin α1 and determine its biological activity effectively, which is suitable for the quality identification of thymosin α1