摘要
为了对伊氏锥虫病的免疫诊断提供有价值的单克隆抗体,笔者通过了3次SP2/0骨髓瘤细胞与伊氏锥虫抗原免疫的BALB/c小鼠脾细胞间的融合试验和多次筛选以及克隆化,建立起7株分泌单克隆抗体的杂交瘤细胞株。对其中ⅡG_6杂交瘤作了进一步的检定分析,用ⅡG_6上清液对T.e.抗原进行ELISA试验,显示出高度的特异性,与血吸虫、弓形虫抗原均未出现交叉反应。ELISA测试ⅡG_6细胞株培养上清效价为1:5.1×10~3,而杂交瘤细胞接种BALB/c小鼠诱生腹水抗体的效价为1:1.6×10~7。该株单克隆抗体免疫球蛋白的类型鉴定属IgG_1亚类。该株细胞已在体外培养下,稳定地分泌特异性抗体至少达6个月之久,同时在液氮冻存近8个月之后仍能保持分泌抗体的能力。
In order to supply valuable monoclonal antibodies for immunodiagnosis , through, three fusions between SP2/0 mice myeloma cells and spleen cells of BALB/c mice immunized with the cellular or soluble antigens of Trypano-soma evansi (T.e.) and a lot of screenings and clonings, 7 cell lines of hybri-doma secreting monoclonal antibodies against T. e. have been established. One of the hybridomas, Ⅱ G8, was selected for further analysis. With regular ELISA, culture supernatant of hybridoma ⅡG6 was tested for the specificity against T, e. Results revealed that ⅡG8 McAb had higher species specificity and did not cross react with Schistosoma japonicum or Toxoplasma. ELISA test showed that the titer of culture supernatant of hybridoma ⅡG8 was 1 : 5.1× 103 and the titer of ⅡG8 McAb in the ascites fluid from the BALB/c mice inoculated with the hybridoma cells was as high as 1:1.6×107. By immuno-diffusion with class specific and subclass specific goat antisera ( gout anti-mouse IgG1, IgG2 , IgG2b, IgG3 and IgM ), the immunoglobulin subclass of ⅡG8 monoclonal antibody was identified as IgG1. This cell line was stable to secrete monoclonal antibodies against T.e. by tissue culture at least for six months and maintained the ability of secreting monoclonal antibodies as preserved in liquid nitrogen at least for eight months.
出处
《江西科学》
1992年第3期147-156,共10页
Jiangxi Science
关键词
伊氏锥虫
抗原
单克隆抗体
杂交瘤
Trypanosoma evansi
T.e. Ag ( Antigen of T.e. )
Monoclonal antibodies
ELISA
