胸腺体液因子(THF-γ2)的合成和活性测定

 目的化学合成胸腺体液因子(THF-γ2),并建立纯化、分析和活性测定的方法。方法采用Fmoc固相合成法(SPPS)合成了胸腺体液因子,用中压液相(MPLC)反相柱一步纯化,经HPLC分析纯度,通过质谱和氨基酸序列测定鉴定合成产物。根据THF-γ2促进T细胞产生IL-2的特性检测合成产物的体外活性。结果化学合成胸腺体液因子的产率为95.8%。纯化后其纯度达到95.92%。质谱测定其相对分子质量为918,与理论相对分子质量相符。氨基酸序列测定为NH₂/Leu-Glu-Asp-Gly-Pro-Lys-Phe-Leu,与设计序列一致。当THF-γ2浓度为300μg·L^-1时,小鼠脾细胞产生IL-2的浓度最高,为空白对照的1.9倍。结论在国内首次建立了胸腺体液因子(THF-γ2)从合成、纯化、分析到活性测定的方法,为胸腺体液因子的进一步研究和应用打下了基础。

Synthesis and Activity Determination of Thymus Humoral Factor (THF- γ2)

OBJECTIVE To study the synthesis,purification,analysis and activity determination of Thymus Humoral Factor(THF-γ2).METHODS Thymus Humoral Factor(THF-γ2) was synthesized by FmocSPPS,purified by MPLC,and identified by HPLC,MS and amino acid sequence determination.The biological activity of THF-γ2 was measured according to its characteristics of promoting IL-2 production in mouse spleen cells. RESULTS The purity of the product was 95.92%,the molecular mass was 918,and the amino acid sequence was NH₂/Leu-Glu-Asp-Gly-Pro-Lys-Phe-Leu.In the concentration of 300 μg·L^-1,the activity was largest and 1.9 times as much as the control.CONCLUSION The methods of the synthesis,purification,analysis and activity determination of Thymus Humoral Factor(THF-γ2) are established for further research and applications in China.