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題 名 | 豬血中類胰島素生長因子第一型之簡易抽取=Simplified Isolation of Insulin-Like Growth Factor-1 from Porcine Blood |
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作 者 | 曹博宏; 吳輔祐; 顏宏達; | 書刊名 | 中國畜牧學會會誌 |
卷 期 | 26:3 1997.09[民86.09] |
頁 次 | 頁337-345 |
分類號 | 473.6 |
關鍵詞 | 豬血; 類胰島素生長因子第一型; 抽取; Porcine blood; IGF-1; Isolation; |
語 文 | 中文(Chinese) |
中文摘要 | 本實驗簡化類胰島素生長因子第一型(Insulin-like Growth Factor-1, IGF-1) 之抽取,以提昇豬血之利用價值並供IGF-1的研究。抽取各步驟IGF-1之含量與活性,分別 以放射免疫分析法及小鼠乳腺上皮(Mouse Mammary Epithelial, MME)細胞培養偵測之。探 集之豬血經冷藏過夜凝集,再離心收集血清,並調pH至2.8,以分離IGF-1與其結合蛋白。 酸化血清離心除沉澱後與SP-Sephadex C-25 離子交換樹酯混合攪拌過夜,經16um孔徑陶瓷 膜過濾,並以pH2.8 溶液清洗後,再回填至管柱。以pH 4.0醋酸氨溶液清洗管柱,再以 再以 pH 9.5 溶液洗出 IGF-1。測得分液以 pH5.2 左右含 IGF-1 最多,達 1.5ug/ml。 這 些分液混合後,經 PM30 超過濾膜除掉大於 30KD 分子後, 再以 YM1 膜濃縮,此步驟增加 IGF-1 相對濃度至 1.2ug/mg 蛋白質。 經等電點分離器 (Rotofor Cell) 兩次分離後, pH8.5 左右之分液 IGF-1 相對濃度最高,可達血液的 17,000 倍以上。 |
英文摘要 | This experiment was conducted to simplify IGF-1 isolation from porcine blood for research purposes and to increase animal byproduct utilization. The concentration and bio-activity of IGF-1 in each isolation step were monitored by radioimmunoassay and MME cell culture, respectively. Blood was coagulated overnight and centrifuged to collect serum. The pH of the serum was adjusted to 2.8 to separated IGF-1 from its binding proteins. After centrifugation, the supernatant was mixed with SP-Sephadex C-25 resin and stirred overnight. The resin was collected and washed by 16 um pore size porcelain filter, and then loaded to column. The column was further washed with pH 4.0 ammonium acetate solution, and then, IGF-1 was eluted with pH 9.5 solution. The highest IGF-1 concentration, up to 1.5 u g/ml was detected around pH 5.2 fractions. The pH 4.5 to 6.0 fractions were pooled and filtered through PM30 ultrafiltration membrane to remove molecules larger than 30 KD, and then concentrated with YM1 membrane. This step increased IGF-1 specific concentration to 1.2 u g/mg protein. Further separation using twice of Rotofor iso-electric focusing indicated that pH 8.5 fraction had the highest IGF-1 specific concentration, which was 17,000 times more than that of original blood. |
本系統中英文摘要資訊取自各篇刊載內容。