不同添加因子对杂交瘤细胞生长和抗体分泌的影响  

Effect of Different Additive Factors on the Growth and Antibody Secretion of Hybridoma Cells

作　　者：张松雨 曾馨仪 朱源涛 李志远 苏俊 李秋艳[1] ZHANG Song-Yu;ZENG Xin-Yi;ZHU Yuan-Tao;LI Zhi-Yuan;SU Jun;LI Qiu-Yan(College of Biological Sciences/State Key Laboratory of Animal Biotech Breeding/National Demonstration Center for Experimental Education of Life Science,China Agricultural University,Beijing 100193,China)

机构地区：[1]中国农业大学生物学院/畜禽生物育种全国重点实验室/生命科学国家级实验教学示范中心,北京100193

出　　处：《农业生物技术学报》2025年第2期379-385,共7页Journal of Agricultural Biotechnology

基　　金：国家现代农业产业技术体系(CARS-36)。

摘　　要：杂交瘤细胞培养是单克隆抗体(monoclonal antibody,McAb)生产的重要方法之一。本课题组前期制备出猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)的杂交瘤细胞单克隆抗体,本研究对杂交瘤细胞培养体系进行优化,在杂交瘤细胞培养体系中加入不同浓度的添加因子,包括谷氨酰胺(0~10 mmol/L)、抗菌肽(0~20μg/mL)、人表皮生长因子(human epidermal growth factor,hEGF)(0~20 ng/mL)、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)(0~20 ng/mL)和维生素C(vitamin C,VitC)(0~20 ng/mL),采用生长曲线绘制和ELISA方法探究添加因子对杂交瘤细胞生长和抗体分泌的影响。结果显示,谷氨酰胺和抗菌肽添加在第5天达到细胞密度的峰值,其中细胞密度最大的添加浓度为6 mmol/L谷氨酰胺和10μg/mL抗菌肽;hEGF、bFGF和VitC添加在第4天达到细胞密度的峰值,其中细胞密度最大的添加浓度为10 ng/mL hEGF、20 ng/mL bFGF和20 ng/mL VitC。抗体效价检测结果显示,谷氨酰胺添加组在6 mmol/L浓度获得最高的抗体效价(1∶8000),抗菌肽添加组在10μg/mL浓度获得最高的抗体效价(1∶16000),hEGF和bFGF添加组在5和10 ng/mL浓度均具有最高的抗体效价(1∶16000),VitC在10 ng/mL浓度获得最高的抗体效价(1∶8000)。本研究通过改变培养基成分提高杂交瘤细胞增殖能力和抗体分泌能力,为研发生产用杂交瘤细胞培养基提供技术支持。The hybridoma cell technique is a crucial method for monoclonal antibody production.Monoclonal antibody-producing hybridoma cells against Porcine epidemic diarrhea virus(PEDV)have been prepared in this research group.In this study,the hybridoma cell culture system was optimized to improve the efficiency of antibody secretion from hybridoma cells by adding different concentrations of additive factors,including glutamine(0~10 mmol/L),antimicrobial peptides(0~20μg/mL),peptides(0~20 ng/mL),human epidermal growth factor(hEGF)(0~20 ng/mL),basic fibroblast growth factor(bFGF)(0~20 ng/mL),and vitamin C(VitC)(0~20 ng/mL).These factors were introduced into the hybridoma cell culture system to investigate their impact on the growth and antibody secretion of hybridoma cells by plotting growth curves and using enzyme-linked immunosorbent assay(ELISA),respectively.The results showed that the peak of cell density was reached on the 5th day when glutamine and antimicrobial peptides were added,and the maximum cell density was achieved at the addition concentration of 6 mmol/L glutamine and 10μg/mL antimicrobial peptides.The peak of cell density was reached on the 4th day when hEGF,bFGF and VitC were added,and the maximum cell density was achieved at the addition concentration of 10 ng/mL hEGF,20 ng/mL bFGF and 20 ng/mL VitC.The results of antibody titer detection showed that the highest antibody titer(1∶8000)was obtained in the glutamine addition group at a concentration of 6 mmol/L,the highest antibody titer(1∶16000)was obtained in the antimicrobial peptide addition group at a concentration of 10μg/mL,the highest antibody titer(1∶16000)was obtained in the hEGF and bFGF addition groups at concentrations of 5 and 10 ng/mL,respectively,and the highest antibody titer(1∶8000)was obtained in the VitC addition group at a concentration of 10 ng/mL.In conclusion,the growth and antibody production capabilities of hybridoma cells have been effectively enhanced through the modification of medium components.This study establishes

关 键 词：杂交瘤细胞 谷氨酰胺 抗菌肽 人表皮生长因子(hEGF) 碱性成纤维细胞生长因子(bFGF) 维生素C 

分 类 号：S828[农业科学—畜牧学] Q813.1[农业科学—畜牧兽医]