摘 要：
采用纯化的鸡IgG免疫环磷酰胺预处理BALB/c小鼠后,再用亲和层析纯化的鸡IgM进行免疫,运用淋巴细胞杂交瘤技术,以间接ELISA法筛选,获得了5株能稳定地分泌抗鸡IgMμ链特异性单克隆抗体的杂交瘤细胞株。经ELISA交叉反应性试验和羊抗鸡IgMμ链抗血清阻断试验,证明这5株单克隆抗体均是抗鸡IgMμ链特异性的。采用环磷酰胺免疫抑制法制备单克隆抗体,其杂交瘤细胞阳性率为8.696％,而用常规免疫法为4.032％,前者比后者杂交瘤细胞阳性率提高1倍以上。经两次有限稀释法克隆和比较后,免疫抑制法获得5株单克隆抗体,而常规免疫法只获得2株单克隆抗体,前者有3株单克隆抗体腹水ELISA效价达到10~(-12)、10~(-14)、10~(-15);后者单克隆抗体腹水ELISA效价为10~(-5)和10~(-6)。因此,与常规免疫法相比,环磷酰胺免疫抑制法制备单克隆抗体具有杂交瘤细胞阳性率高,分泌的单克隆抗体特异性强、滴度高等优点,值得推广。
译 名：
Development of Monoclonal Antibodies with Immunosuppressive Pretreated with Cyclophosphamide
作 者：
Qian Jianfei Mao Hongxian Shao Guoqing Jin Hongxiao Lli Lixin(Institute of Animal Husbandry and Veterinary Science, Jiangsu Academy of Agricultural Sciences?Nanjing 210014)
关键词：
Chicken IgM; Cyclophosphamide; Monoclonal antibody
摘 要：
Five hybridoma cell lines were obtained which secrete antibodies specific for chicken IgM (μ chain) by fusion of SP2/0 myeloma cells with splenocytes from BALB/c mice immunized with purified chicken IgM by affinity chromatography from chicken serum, after pretreated of adult mice with chicken IgG and cyclophosphamide. All monoclonal antibodies (McAbs) were specific for μ chain of chicken IgM by cross-reactivity test with indirect ELISA and ELISA blocking test with serum of goat anti-μ chain of chicken IgM . The positive rate of hybridoma cells was respectively 8. 696% and 4. 032% with im-munosuppressive pretreat with cyclophosphamide and normal immunization. The positive rate of the former was twice more than that of the latter. After cloned twice with limited dilution and compared, five McAbs and two McAbs were respectively obtained by im-munosuppressive and normal immunization methods. The ELISA liter of three McAbs were above 10-12 in ascites in the former, and the highest titer reached 10-15 in ascites. However the ELISA titer of two McAbs were 10-5 and 10-8 in ascites in the latter. Therefore , immunosuppressive pretreated with cyclophosphamide had higher positive rate of hybridoma cells, stronger specificity of McAbs and higher titer of McAbs than normal immunization when McAbs were developed. It was worth spreading and applying that the immunosuppressive method was pretreated with cyclophosphamide by McAb researchers.
