固定pH梯度等电聚焦法分离纯化人胸腺活性肽

 本文用4—7月龄胎儿胸腺,按照匀浆—80℃加热—丙酮沉淀—抽提—超滤等一系列步骤制备人胸腺混合肽。用固定pH梯度聚丙烯酰胺凝胶等电聚焦技术,结合紫外薄层扫描法定位,从胸腺混合肽中得到四种纯化的胸腺肽HTP α_1,β_1,β_2和γ_1;分别测得其等电点为4.6,5.3,5.8和7.6,分子量分别为4800,6700,7200—7300和12,000—13,000。另外至少还得到四种部份纯化的胸腺肽。由玫瑰花结和末端转脱氧核苷酰酶活力测定结果,证明分离得到的各种胸腺肽均有较高的生物活性。

PURIFICATION OF HUMAN THYMIC PEPTIDES BY PREPARATIVE ELECTROFOCUSING IN IMMOBIL pH GRADIENTS

An efficient and simple method for purification of human thymic peptides (HTP) by preparative electrofocusing in immobil pH gradients is recommended. The fresh human fetus tissues were collected and treated in steps by heat denatu-ration, acetone precipitation and ultrafiltration. The ultrafiltrate (mHTP) was separated by preparative electrofocusing in immobil pH gradients. Four peptides, HTPα,β,β2 and γ1 in mg level were successfully purified from one gel plate with dimensions 110 ×240× 2mm. The isoelectrie points of these purified peptides are 4.6, 5.3. 5.8 and 7.6, and the molecular weights estimated by SDS-PAGE are 4,800, 6,700, 7,200-7,300 and 12,000-13,000 dalton respectively. The four purified HTPs were shown to maintain high activities in both E-rosette assay and TdT activity test.