血液因子动物生物反应器研制与产品开发课题报告

摘 要：该项目旨在利用乳腺生物反应器成功获得目标蛋白的基础上进行深入研究，综合应用分子生物学、细胞学、胚胎学、繁殖学、育种学、生物化学、药理学等多学科手段研究动物生物反应器所生产药物蛋白的有效性和安全性。将围绕目标蛋白的临床前研究中的中试批量质控生产、药理毒理学等进行研究，从而使国内首个转基因动物生产的新型生物药品的开发迈进重要一步。该研究课题获得了高表达且稳定遗传的牛和羊乳腺生物反应器，且抗凝血酶III转基因阳性羊种群规模已达81只，转基因牛种群规模达到15头；将AT-Ⅲ纯化工艺进行逐级放大，目前能够达到30 L/批次，正在尝试进行放大到50 L/批次，蛋白纯度达到98%以上；采用两种方案对hFIX蛋白进行纯化，方案一的比活性和纯度高于第二种方案，但方案二的回收率较高；经过对纯化的rhATⅢ和hFⅨ蛋白的结构进行确证，均和人源的相似；通过药物制剂的研究，rhATⅢ蛋白确定了合适的制剂配方；目前正在进行药品注册临床前研究工作。

关键词：动物乳腺生物反应器 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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