Big-BALB/c小鼠亚系选育前后的抗体制备效率比较研究

Study on the Antibody Production Efficiency in Modified Big-BALB/c Mice

目的比较利用BALB/c和Big-BALB/c(简称B-BALB/c)两种亚系小鼠制备鼠痘及鼠肝炎单克隆抗体的效率,为今后通过杂交瘤进行体内诱导单克隆抗体制备时实验动物的选择提供参考。方法从常规繁育的BALB/c小鼠群体中选择4周龄体重超过正常动物5%(后期选种标准为超过10%)的个体进行选种并单独繁育,经10代选育制备B-BALB/c小鼠亚系。选择10~11周龄、雌雄各半的BALB/c小鼠和B-BALB/c小鼠各40只,分别接种用液体石蜡预处理后的鼠痘单克隆抗体杂交瘤细胞株G23或鼠肝炎单克隆抗体杂交瘤细胞株Y15,然后通过体内诱生法获取含单克隆抗体的小鼠腹水,利用间接ELISA法检验抗体效价,按照品系、性别和接种杂交瘤类型对小鼠进行分组,分析腹水产量、抗体效价和抗体产量以评估两种亚系小鼠的抗体制备效率。结果经10代选育后获得的10周龄雄性和雌性B-BALB/c小鼠体重分别比同周龄BALB/c小鼠增加了22.3%和12.8%。抗体制备过程中,B-BALB/c小鼠的耐受力比BALB/c小鼠更好,能够适应二次腹水采集;与BALB/c小鼠相比,B-BALB/c小鼠腹水产量和抗体效价均显著提高,尤以杂交瘤细胞株G23接种组更明显(均P<0.0001)。接种杂交瘤细胞株G23或Y15后,B-BALB/c小鼠的平均抗体产量(14.99×10^(4)U和33.22×10^(4)U)高于BALB/c小鼠(5.33×10^(4)U和19.31×10^(4)U)(均P<0.01)。接种杂交瘤细胞株G23后,B-BALB/c小鼠的平均单位体重的抗体产量(0.55×10^(4)U/g)高于BALB/c小鼠(0.23×10^(4)U/g)(P<0.0001)。接种杂交瘤细胞株Y15后,雌性B-BALB/c和BALB/c小鼠的单位体重抗体产量高于同亚系的雄性B-BALB/c小鼠(均P<0.01)。结论B-BALB/c小鼠在单克隆抗体体内诱导制备中可作为BALB/c小鼠的替代选择,能够达到减少实验动物使用数量、降低人力成本并提高抗体制备效率的目的。

Objective To compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c(B-BALB/c)mice,and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies induced in vivo through hybridoma.Methods Individuals weighing more than 5%of the weight of normal animals at 4 weeks of age(the criterion for late selection is more than 10%)were selected from a population of conventionally bred BALB/c mice and bred individually,and a subline of B-BALB/c mice was prepared after 10 generations of selection.A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks,half male and half female,were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin,respectively.Mice ascites containing monoclonal antibodies were obtained by in vivo induction.The antibody titer was tested by indirect ELISA.The mice were grouped based on the sub-strains,gender and inoculation type of hybridoma to analyze the ascites production,antibody titer and antibody production,and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains.Results After 10 generations of breeding,the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3%and 12.8%,respectively,compared with BALB/c mice of the same age.Compared with BALB/c mice,B-BALB/c mice had better tolerance and adaptation to secondary ascites collection.Compared with BALB/c mice,the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased,especially in the hybridoma cell line G23 vaccination group(both P<0.0001).After inoculation with the hybridoma cell lines G23 o r Y 15,the average antibody production of B-BALB/c mice(14.99×10^(4)U and 33.22×10^(4)U)was higher than that of BALB/c mice(5.33×10^(4)U and 19.31×10^(4)U)(both P<0.01).After inoculation with hybridoma cell line G23,the average antibody production per unit body weight of B-BALB/c mice(0.55×10^(4)U/g)was higher than that of BALB/c mice(0.23×10^(4)U/g)(P<0.0001).And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice(both P<0.01).Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation,which can achieve the purpose of reducing the number of experimental animals used,lowering the labor cost,and improving the efficiency of antibody preparation.