食品研究与开发
食品研究與開髮
식품연구여개발
FOOD RESEARCH AND CEVELOPMENT
2013年
12期
20-25
,共6页
刘妍妍%刘茗飞%高薇%张丽萍
劉妍妍%劉茗飛%高薇%張麗萍
류연연%류명비%고미%장려평
胸腺肽α1%离子交换色谱%分离纯化
胸腺肽α1%離子交換色譜%分離純化
흉선태α1%리자교환색보%분리순화
Thymosinα1%ion-exchange chromatography%isolation and purification
通过对胸腺肽α1的分离纯化,提高胸腺肽α1的纯度,确定简单可行的纯化工艺技术。将QAE Sephadex A-25和D208、D301、717等阴离子交换介质进行静态吸附实验,确定最适离子交换介质;通过L9(33)正交试验对流速、填料高度及进样浓度进行优化,确定最佳分离条件。以QAE Sephadex A-25作为填料介质吸附胸腺肽α1,在流速60 cm/h,填料高度50 cm,进样浓度8 mg/mL条件下,胸腺肽α1纯化效果理想,经HPLC测定,得到纯度达24.5966 mg/mL的胸腺肽α1,质量分数达12.03%,比市场上销售的胸腺肽中Tα1含量(质量分数0.6%~1.0%)高出近10倍。
通過對胸腺肽α1的分離純化,提高胸腺肽α1的純度,確定簡單可行的純化工藝技術。將QAE Sephadex A-25和D208、D301、717等陰離子交換介質進行靜態吸附實驗,確定最適離子交換介質;通過L9(33)正交試驗對流速、填料高度及進樣濃度進行優化,確定最佳分離條件。以QAE Sephadex A-25作為填料介質吸附胸腺肽α1,在流速60 cm/h,填料高度50 cm,進樣濃度8 mg/mL條件下,胸腺肽α1純化效果理想,經HPLC測定,得到純度達24.5966 mg/mL的胸腺肽α1,質量分數達12.03%,比市場上銷售的胸腺肽中Tα1含量(質量分數0.6%~1.0%)高齣近10倍。
통과대흉선태α1적분리순화,제고흉선태α1적순도,학정간단가행적순화공예기술。장QAE Sephadex A-25화D208、D301、717등음리자교환개질진행정태흡부실험,학정최괄리자교환개질;통과L9(33)정교시험대류속、전료고도급진양농도진행우화,학정최가분리조건。이QAE Sephadex A-25작위전료개질흡부흉선태α1,재류속60 cm/h,전료고도50 cm,진양농도8 mg/mL조건하,흉선태α1순화효과이상,경HPLC측정,득도순도체24.5966 mg/mL적흉선태α1,질량분수체12.03%,비시장상소수적흉선태중Tα1함량(질량분수0.6%~1.0%)고출근10배。
@@@@Through purifying thymosinα1 and improve the purity of thymosinα1 in order to research a simple and feasible purification technology. The study chooses QAE Sephadex A-25 and D208, D301, 717 ect. for static IV adsorption experiment through the anion exchange chromatography in order to determine the optimum ion exchange media;The optimal experiment conditions of the eluent, eluting style, flow rate, height of media are studied and choose L9 (33) orthogonal experiment on optimizing the conditions of the velocity, height and the concentration. The study chooses HPLC and SDS-PAGE to identifying the thymosinα1. The experiment showed that the QAE Sephadex A-25 as a filler media adsorbing thymosinα1 makes the best effect, the velocity is 60 cm/h, packing height is 50 cm, the concentration is 8 mg/mL, then the purification results is satisfactory. Determined by the HPLC, the purity of thymosinα1 is 24.596 6 mg/mL, the mass percent is 12.03%. The method is accurate and feasible. The purity of thymosinα1 is 10 times higher than the commercial thymosinα1 products (the mass percent is 0.6%to 1.0%).