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当前位置: 首页 > 文章 > 喹乙醇检测单抗制备中细胞融合条件的优化 黑龙江畜牧兽医 2013 (3) 81-84+172
Position: Home > Articles > Optimization of cell fusion conditions in preparation of monoclonal antibodies for detecting olaquindox Heilongjiang Animal Science and Veterinary Medicine 2013 (3) 81-84+172

喹乙醇检测单抗制备中细胞融合条件的优化

作  者：
王磊;李建喜;张景艳;王学智;杨志强
单  位：
中国农业科学院兰州畜牧与兽药研究所
关键词：
喹乙醇;单克隆抗体;细胞融合;聚乙二醇;优化
摘  要：
为了优化抗喹乙醇单克隆抗体制备中聚乙二醇(PEG)诱导细胞融合的最适条件,试验采用流式细胞仪确定SP2/0细胞的对数生长期,以融合率和阳性率为评价指标,分析血清加入量、500 mL/LPEG-1000溶液方法及反应时间和次黄嘌呤、氨基蝶呤、胸腺嘧啶脱氧核苷(HAT)加入时间对SP2/0细胞与脾细胞融合效果的影响。结果表明:150,200 mL/L胎牛血清融合率分别为86.5%、88.5%;生理盐水和无血清DMEM配制的500 mL/L PEG-1000融合率和阳性率分别为61.4%、3.1%,30.2%、1.0%;PEG-1000反应1.75,3.00 min融合率和阳性率分别为61.4%、3.1%,74.5%、6.3%;融合后培养0,24 h加入HAT,结果融合率和阳性率分别为95.8%、17.7%,94.3%、26.0%。说明150 mL/L胎牛血清、500 mL/L PEG-1000生理盐水、反应3.00 min、培养24 h后加入HAT时,细胞融合率和阳性率较高,是抗喹乙醇单克隆抗体制备中PEG诱导细胞融合的最适条件。
译  名：
Optimization of cell fusion conditions in preparation of monoclonal antibodies for detecting olaquindox
作  者：
WANG Lei1,2,3,LI Jian-xi1,2,3,ZHANG Jing-yan1,2,3,WANG Xue-zhi1,2,3,YANG Zhi-qiang1,2,3, ZHANG Kai1,2,3,MENG Jia-ren1,2,3 (1.Lanzhou Institute of Husbandry and Pharmaceutics,CAAS,Lanzhou 730050,China;2.Key Laboratory of New Animal Drug Project,Ministry of Agriculture,Lanzhou 730050,China;3.Engineering & Technology Research Center of Traditional Chinese Veterinary Medicine of Gansu Province,Lanzhou 730050,China)
关键词：
olaquindox;monoclonal antibody;cell fusion;polyethylene glycol(PEG);optimization
摘  要：
To optimize the conditions of PEG-induced cells fusion in the preparation of monoclonal antibodies against olaquindox,the logarithmic growth stage of SP2/0 cells was determined with flow cytometry.The effect of the four factors on the cell fusion between the SP2/0 cells and spleen cells was analyzed,by comparing the cell fusion rate and the positive rate,the four factors included fetal bovine serum concentration,the preparation and reaction time of 500 mL/L PEG-1000 and the addition time of hypoxanthine,aminopterin,and thymidine(HAT).The results showed that the fusion rates treated with 150 mL/L and 200 mL/L fetal bovine serum(FBS) were 86.5% and 88.5%,respectively.The fusion rates treated with normal saline and 500 mL/L PEG-1000 with serum-free DMEM were 61.4% and 30.2%,and the positive rates treated with normal saline and 500 mL/L PEG-1000 with serum-free DMEMwere 3.1% and 1.0%,respectively.The fusion rate and positive rate were 61.4% and 3.1% after PEG reacting for 1.75 min,and 74.5% and 6.3% after PEG reacting for 3.00 min,respectively.The fusion rate and positive rate were 95.8% and 17.7% with HAT being added to the culture medium at 0 h after cell fusion,respectively.They were 94.3% and 26.0% with HAT being added at 24 h after cell fusion.In the experiment,the results indicated that the optimal conditions of PEG-induced cells fusion in the preparation of monoclonal antibodies against olaquindox,were 150 mL/L FBS,500 mL/L PEG-1000 in normal saline,the reaction time for 3.00 min,and the addition of HAT to the culture medium at 24 h after the culture.

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