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作 者:王红连 陆丽婷 于婷 匡越 华洵璐 石文婷 朱亚俊 张海涛 WANG Honglian;LU Liting;YU Ting;KUANG Yue;HUA Xunlu;SHI Wenting;ZHU Yajun;ZHANG Haitao(Jiangsu Suwei Microbial Research Co.,Ltd.,Wuxi 214063,China)
机构地区:[1]江苏省苏微微生物研究有限公司,江苏无锡214063
出 处:《生物化工》2024年第5期112-115,共4页Biological Chemical Engineering
摘 要:本研究通过优化离心条件,利用小鼠外周血淋巴细胞分离液成功分离出小鼠腹水中的抗呕吐毒素(DON)杂交瘤细胞SW-3H_(2),并考察不同周龄和性别小鼠的腹水产量及SW-3H_(2)在腹腔传代中的稳定性。结果表明,2000 r/min、20 min的离心条件可获得最佳分离效果。分离获得的杂交瘤细胞可再次接种10~15只小鼠,且在14周龄雄性小鼠中腹水产量显著高于雌性小鼠。此外,杂交瘤细胞在小鼠腹腔中可连续稳定传代3代,平均腹水产量为5.95 mL/只,抗体效价超过1∶4.9×10^(4)。本研究的成果将有助于提高抗DON单克隆抗体的制备效率。This study optimizes the centrifugation conditions to successfully separate anti-vomitoxin(DON)hybridoma cells(SW-3H_(2))from mice ascites using mice peripheral blood lymphocyte separating fluid.It also investigates the abdominal fluid production of these cells in mice of different ages and genders,as well as their stability during intraperitoneal passaging.The results demonstrate that the optimal separation efficiency is achieved with centrifugation at 2000 r/min for 20 mins.The separated hybridoma cells could be re-inoculated in 10 to 15 mice,and the ascites production is significantly higher in 14-week-old male mice than in female mice.Furthermore,the hybridoma cells show stable passage for three generations in the mice peritoneal cavity,with an average ascites yield of 5.95 mL per mouse and antibody titer exceeding 1 to 4.9×10^(4).The findings of this study are expected to enhance the efficiency of preparing anti-DON monoclonal antibodies.
关 键 词:抗呕吐毒素杂交瘤细胞 小鼠外周血淋巴细胞分离液 小鼠腹水
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