牛布鲁菌O链A抗原的提纯鉴定及种特异性单克隆抗体的研制

Purification and identification of B.abortus 544A O chain-A antigen and preparation of the strain specific monoclonal antibodies

应用热酚法提纯牛布鲁菌544A的O链,经Sephades G-50纯化后,琼脂糖凝胶免疫扩散和SDS聚丙烯酰胺凝胶进行电泳鉴定,用牛布鲁菌544A株全菌体免疫BALB/c小鼠,用纯化的O链筛选出4株针对O链表面A抗原的种特异性单克隆抗体的杂交瘤细胞株4B8、1C9、1G9和1E2,4株单克隆抗体均不与羊布鲁菌16 M、大肠杆菌O157、小肠结肠炎耶尔森菌O9、鼠伤寒沙门菌、放线杆菌发生交叉反应,具有高特异性,其中4B8的亲和常数达到了8.99×108M-1,适合用于鉴定牛种布鲁菌的研究。

O chain of B. abortus 544A was purified with heat-phenol and identified with agarose immunodiffusion （AGID） and SDS polyacrylamide gel electrophoresis（SDS-PAGE）. A panel of four hybridoma cell lines secreting strain specific antibodies against O chain-A antigen of B. abortus 544A were established with purified O chain. The four monoclonal antibodies（McAbs） are all high specificity because they don＇t react with B. melitensis 16 M,E. coli O157, Y. enterocolitica O9, S. typhimurium, Actinobacillus etc. And 4B8 have high affinity quantity of 8. 99 × 10^8 M^-1 , so it is suitable to be used to study the method for detection and identification of the B. abortus.