表达抗鸡传染性法氏囊病病毒重组鸡源抗体CHO细胞的悬浮驯化  

Suspension domestication of the CHO cells expressing recombinant chicken anti-chicken infectious bursal disease virus antibody

作　　者：曹宏雪 郭笑辰 李婷婷[1] 张胜奇 张瑛杰[1] 刘鑫[1] 任桂萍[1,2] 尹杰超[1,2] Lubna Muhi Rasoul 荆燕[1,2] 李德山[1,2] 

机构地区：[1]东北农业大学生命科学学院/生物制药教研室,黑龙江哈尔滨150030 [2]东北农业大学生命科学学院/生物功能基因重点实验室,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2016年第1期68-72,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省应用技术研究与开发计划项目(GC13C104);黑龙江省博士后科研启动基金(LBH-Q09162)

摘　　要：为将贴壁生长的表达抗鸡传染性法氏囊病病毒(IBDV)重组鸡源抗体的CHO细胞株驯化为在无血清培养基中悬浮生长、高产的工程细胞株,本研究利用流式细胞仪筛选得到高产贴壁细胞株;以快速降低血清法进行驯化,使其适应悬浮培养,经过摇瓶无血清培养扩增,持续监测葡萄糖消耗情况,最终检测抗IBDV抗体的表达量。结果显示,悬浮驯化培养的CHO细胞能够在无血清培养基中稳定生长,生产周期短,接种密度为4.8×10~5cells/m L,无血清单细胞悬浮培养120 h后最大活细胞密度可达6.0×10~6 cells/m L,相同体积下抗体表达量较贴壁生长的高3倍,平均每个细胞每24 h表达量达到15 pg^20 pg。以上结果表明,贴壁生长的CHO细胞经过悬浮适应,不仅可以在无血清培养条件下快速生长,而且细胞对葡萄糖的利用率高,抗体表达量高。本研究筛选获得的细胞株节省了工艺生产中原料成本,操作方便,减少污染,易于放大,为工业化大批量生产奠定了基础。This study aims to adapt the expression of recombinant anti-chicken infectious bursal disease virus （IBDV） chicken antibodies in adhesive growth of CHO cell lines domesticated in serum free medium to yield strains of highly engineered cells for future manufacturing production. Adherent growth of CHO cells expressing the recombinant chicken anti-IBDV antibodies was acclimated to suspension growth in serum free medium. Using rapid reduction of serum for domestication makes it adapt suspended culture in serum free culture flask after amplification. Cells were compared before and after adoption for their antibody production. Glucose consumption situation and affinity were daily monitored; eventually anti-IBDV antibody expression, neutralization and specificity ability were also detected. Results showed that domesticated suspension of CHO cells cultured can grow in serum free medium in short production cycle. The maximum density of the domesticated cells reached 6.0×10^6 cells/mL after inoculated at an initial density of 4.8×10^5 cells/mL after 120 h of bloodless cell suspension culturing. The antibody yield of the domesticated culture reached 15-20 pg/day/cell, which was three times higher than that of the adherent culture under the same condition. These results indicate that adherent growth of CHO-IBDV cells after suspension adapts not only rapid growth in serum-flee culture conditions, but also increases rate of glucose utilization and increases antibody production then the cells become ready for antibody production in industrial level. The advantages of screened cell line production processes is raw materials cost saving, ease of operation, reduced pollution, easy to enlarge, and create a basis for industrializing mass production of antibodies against IBDV.

关 键 词：传染性法氏囊病病毒 悬浮驯化 CHO 鸡源抗体 

分 类 号：S852.65[农业科学—基础兽医学]