血液因子動物生物反應(yīng)器研制與產(chǎn)品開發(fā)課題報告

趙雅琳 曾凡偉

摘 要：該項目旨在利用乳腺生物反應(yīng)器成功獲得目標蛋白的基礎(chǔ)上進行深入研究，綜合應(yīng)用分子生物學(xué)、細胞學(xué)、胚胎學(xué)、繁殖學(xué)、育種學(xué)、生物化學(xué)、藥理學(xué)等多學(xué)科手段研究動物生物反應(yīng)器所生產(chǎn)藥物蛋白的有效性和安全性。將圍繞目標蛋白的臨床前研究中的中試批量質(zhì)控生產(chǎn)、藥理毒理學(xué)等進行研究，從而使國內(nèi)首個轉(zhuǎn)基因動物生產(chǎn)的新型生物藥品的開發(fā)邁進重要一步。該研究課題獲得了高表達且穩(wěn)定遺傳的牛和羊乳腺生物反應(yīng)器，且抗凝血酶III轉(zhuǎn)基因陽性羊種群規(guī)模已達81只，轉(zhuǎn)基因牛種群規(guī)模達到15頭；將AT-Ⅲ純化工藝進行逐級放大，目前能夠達到30 L/批次，正在嘗試進行放大到50 L/批次，蛋白純度達到98%以上；采用兩種方案對hFIX蛋白進行純化，方案一的比活性和純度高于第二種方案，但方案二的回收率較高；經(jīng)過對純化的rhATⅢ和hFⅨ蛋白的結(jié)構(gòu)進行確證，均和人源的相似；通過藥物制劑的研究，rhATⅢ蛋白確定了合適的制劑配方；目前正在進行藥品注冊臨床前研究工作。

關(guān)鍵詞：動物乳腺生物反應(yīng)器 抗凝血酶Ⅲ 凝血因子Ⅸ

Abstract：This project is intend to deeply study the mammary gland bio- reactor on the basis of successfully getting the target protein， and comprehensively using molecular biology， cytology， embryology， reproductive science， thremmatology， biochemistry， pharmacology and other subjects to research the efficacy and safety of pharmaceutical proteins produced by animal bioreactor。We will focus on researching the pilot batch quality control、pharmacology and toxicology about target protein，which are required to be studied in preclinical，so the development of the new biological drug produced by Transgenic animal will be an important step forward. For this project，we obtained mammary gland bioreactor of transgenic cattle and goats with high and stable expression。And we have transgenic goats and 15 transgenic cattle； We will enlarge AT-Ⅲ purification proce- ss progressively. At present，the purification scale can reach 30 L/batch， we try to enlarge it to 50 L/batch； protein purity reach more than 98%. we use two solutions to purify the hFIX protein， specific activity and purity of the first solution are higher than the second solution， but the second solution get higher recovery rate；After confirming the structure of proteins，the purified rhAT-Ⅲ and hF-Ⅸ are similar with human sources；We determined a suitable formula of the preparation about rhAT-Ⅲ through researching the pharmaceutical prepara- tion；Now，we are working on pre clinical about drug registration。

Key Words：Mammary Gland Bioreactor；AntithrombinⅢ；Blood Coagulation factorⅨ

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