本文選題：EPO + 貧血��； 參考：《吉林大學》2005年博士論文

【摘要】：紅細胞生成素(EPO,erythropoietin)是一種刺激骨髓造血的糖蛋白類激素,基因工程方法生產(chǎn)的重組人紅細胞生成素在治療腎性貧血及其它類型的貧血等方面有很好的療效。 為了增加其生物活性,及延長其半衰期,作者構建了三種不同啟動子的重組人紅細胞生成素(rhEPO)二聚體真核表達載體pBT-1c、pBT-2s、pBT-3c,及兩個rhEPO 單體表達載體pBT-2se,pBT-3ce。比較其轉染效率后,選擇一種真核表達載體pBT-2s 穩(wěn)定轉染CHO-dhfr-細胞,經(jīng)MTX加壓,篩選到穩(wěn)定高效表達rhEPO 二聚體融合蛋白的細胞株CHO-BTE,表達量達到33.3μg /106cell?d。經(jīng)純化得到rhEPO 二聚體融合蛋白,經(jīng)檢測分子量約為70KD,其體內(nèi)及體外生物活性均顯著高于rhEPO 單體。經(jīng)大鼠體內(nèi)的初步藥代動力學試驗,結果表明,此rhEPO 二聚體融合蛋白在大鼠體內(nèi)的半衰期比rhEPO 單體半衰期延長。
[Abstract]:Erythropoietin (EPO) is a glycoprotein hormone that stimulates bone marrow hematopoiesis. Recombinant human erythropoietin produced by genetic engineering has a good effect in the treatment of renal anemia and other types of anemia. In order to increase its biological activity and prolong its half-life, the authors constructed three recombinant human erythropoietin (rhEPO) dimer eukaryotic expression vectors pBT-1, pBT-2spBT-3cand two rhEPO monomeric expression vectors pBT-2sesepBT-3ce. After the transfection efficiency was compared, a eukaryotic expression vector pBT-2s was selected to transfect CHO-dhfr- cells stably. The cell line CHO-BTE-1, which expressed rhEPO dimer fusion protein stably and efficiently, was screened by MTX pressurization. The expression level reached 33.3 渭 g / 10 6 cell line. RhEPO dimer fusion protein was purified and its molecular weight was about 70 KD.The bioactivity of rhEPO was significantly higher than that of rhEPO monomer in vivo and in vitro. The preliminary pharmacokinetic test in rats showed that the half-life of rhEPO dimer fusion protein in rats was longer than that of rhEPO monomer.
【學位授予單位】：吉林大學
【學位級別】：博士
【學位授予年份】：2005
【分類號】：R346

【參考文獻】

相關期刊論文 前1條

1 高云莉;EPO的市場現(xiàn)狀及展望[J];中國新藥雜志;2001年02期

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本文編號：2099300