制备可溶性肿瘤坏死因子受体Ⅱ-脂联素球部融合蛋白的两种细胞培养工艺比较

来源 :生物工程学报 | 被引量 : 0次 | 上传用户:huachao198977
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
利用7.5 L生物反应器篮式贴壁培养和全悬浮批式培养CHO工程细胞株表达可溶性肿瘤坏死因子受体Ⅱ-脂联素球部(sTNFRII-gAD)融合蛋白,比较这两种培养方法的产率,以便优化高效表达sTNFRII-gAD融合蛋白的制备工艺。篮式贴壁培养首先小规模培养CHO工程细胞株,待细胞增殖到一定密度后以3×105~4×105cells/mL密度接种生物反应器贴壁培养3 d,调换成不含血清的LK021培养基继续培养4 d。而全悬浮无血清批式培养则以3×105~4×105cells/mL密度的CHO工程细胞株接种于生物反应器,连续培养7 d。培养过程实时监测培养条件,维持pH和DO的稳定。分别收集细胞上清,离心去细胞后用Pellicon切相流超滤系统对蛋白进行浓缩,并通过DEAE离子交换柱进行纯化。结果显示,篮式贴壁培养和全悬浮批式培养均成功表达了sTNFRII-gAD融合蛋白,产量分别为8.0 mg/L和7.5 mg/L、纯度分别为95%和98%,从而为sTNFRII-gAD融合蛋白的中试工艺研究提供了一定的基础。 The soluble tumor necrosis factor receptor-II-adiponectin (sTNFRII-gAD) fusion protein was expressed in basket-cultured and suspension-fed batch culture CHO cells by 7.5 L bioreactor. In order to optimize the preparation process for the efficient expression of the sTNFRII-gAD fusion protein. Basket Affinity Culture First, CHO cell lines were cultured in small scale. After the cells had proliferated to a certain density, the cells were inoculated with 3 × 105 ~ 4 × 105cells / mL bioreactor for 3 days at a density of 3 × 105 cells / mL and switched to serum-free LK021 culture Base continue to cultivate 4 d. The whole suspension serum-free batch culture with 3 × 105 ~ 4 × 105cells / mL density of CHO engineering cell lines inoculated in the bioreactor, continuous culture for 7 days. The culture process monitors the culture conditions in real time to maintain pH and DO stability. The cell supernatants were collected, centrifuged to remove the cells, then the protein was concentrated by Pellicon phase-flow ultrafiltration and purified by DEAE ion exchange column. The results showed that the sTNFRII-gAD fusion protein was expressed successfully in both basket-type and suspension-type batch cultures with yields of 8.0 mg / L and 7.5 mg / L, respectively, with 95% and 98% purity, respectively, gAD fusion protein pilot process provides a basis for the study.
其他文献
探讨在图书馆人制管理中,“人”与“制”存在着辩证关系,并就“人制管理”内涵的辩证关系从以人定制、以制管理、以制管人、人才管理四个方面进行探讨。 To discuss the exi
研究了基体效应、参比强度K值、衍射强度、物相粒度、X射线管功率等实验条件对TOPAS定量分析的影响。对比了D8 ADVANCE型X射线衍射仪EVA和TOPAS 2种定量分析软件,发现TOPAS定
采用微波消解技术,电感耦合等离子体发射光谱法(ICP-OES)测定化妆品中钛含量。实验中消解体系为6.0 m L浓硝酸、1.0 m L过氧化氢和1.0 m L氢氟酸,方法的检出限为6.0 mg/kg,相
探明影响甲基红光谱性质的各种因素,有助于拓宽偶氮苯衍生物在有机光电器件中的应用。采用密度泛函理论和实验相结合的方法研究了溶液酸碱性和溶剂水对邻位甲基红水溶液光谱
请下载后查看,本文暂不支持在线获取查看简介。 Please download to view, this article does not support online access to view profile.
期刊
竣工于1966年的法国朗斯潮汐电站,装机240 MW,运行方式为双向型,水轮机机组为灯泡式,其土建工程和机电工程技术相当成熟。介绍了该电站多年来的成功运行经验、电站的主要技术
多年老厂,诚信做生意,放心邮寄,中国专利ZL200720150520.6,注册商标:梁新,假一奖你一万元。  市场行情:物价越来越高,买的用品也就会贵,一旦坏了就会修理,你家会这样想到,另外一些家庭也同样会这样去做,焊补的多了,你每天都会有做不完的生意,还有好多用品同样也会坏,他们也同样会找你焊补,这样一来,你地又没有别人会焊补这些用品,你是做独家生意,你想想,到那时候,你不想赚钱都是不行的,因为
患者女,27岁,婚后曾3次妊娠,均在孕2~3个月之间无明显诱因阴道流血,B超检查显示胚胎宫内死亡.患者妇科检查正常,夫妇体健,表型及智力均正常,非近亲结婚,无不良因素接触史.  细胞遗传学检查:取患者外周血淋巴细胞培养,常规方法制片,G显带分析,镜下计数40个中期分裂相,分析15个,患者核型为45,XX,t(21;22)(21qter-cen-22qter),其夫核型正常.
请下载后查看,本文暂不支持在线获取查看简介。 Please download to view, this article does not support online access to view profile.
黄金分割率可以说是世人皆知,其基本公式就是把1分割成0.618和0.382,并且依此再推导和演变出其它的计算公式。 证券投资专家通过对股票市场,期货市场的分析和研究,发现股价