摘要
首先利用弗式完全佐剂和弗式不完全佐剂,乳化病毒,制备免疫抗原,对BALB/c小鼠进行三次免疫。将免疫效价达到1：10000以上的免疫小鼠脾细胞与SP2/0细胞进行融合,通过间接ELISA方法对阳性杂交瘤细胞株进行筛选,并通过有限稀释法将呈强阳性的杂交瘤细胞亚克隆3~4次,直到杂交瘤细胞阳性率达到100%。得到阳性株命名为2E3,对阳性株细胞进行扩大培养,并将细胞注入腹腔,提取腹水,测定细胞上清及腹水的效价,分别为10×25,10×26。利用亚类鉴定试剂盒测定单抗的亚类。鉴定结果为IgG1型。
In order to make DHV antigens,the experiment utilize IFA and CFA. The BALB/c mice were immunized with DHV for three times. The spleen ’ s B lymphocyte of the mice should be fused with SP2/0 plasmacytoma cells until the immunopotency was up to 1：10000. The masccline hybridoma cells were screened by indirect enzyme-link immunosorbent assay. Hybridomas whose supernatants show stronger positive were subcloned by limited dilution methods for 3~4 times, until the positive rate come up to 100%. It was named 2E3. The immunopotency of ascites and supernatant was 10×2 5 ,100×2 6. . And the subtype of 2E3 was IgG l .
出处
《中国农学通报》
CSCD
北大核心
2011年第1期415-418,共4页
Chinese Agricultural Science Bulletin
基金
吉林省科技平台建设项目"吉林省实验动物质量检测中心平台建设"(20071138)
