摘要
用淋球菌全菌体免疫ＢＡＬＢ／ｃ小鼠，取脾细胞与Ｓｐ２／０骨髓瘤细胞融合，以纯化ＰＩＡ做ＥＬＩＳＡ间接法筛选，获得５株稳定分泌抗ＰＩＡ的ＭｃＡｂ的杂交瘤细胞株。５株ＭｃＡｂ中３株为ＩｇＭ类（２Ｈ１１，４Ｈ８，４Ｅ１０），另２株分别为ＩｇＧ１（１Ｃ２）和ＩｇＧ２ｂ（５Ａ５）亚类。２Ｈ１１和１Ｃ２为高亲和力抗体，１Ｃ２与５Ａ５识别的抗原表位相同。Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ试验表明，５株ＭｃＡｂ均能从复杂的淋球菌菌体崩解物中特异地识别分子量为３５ＫＤａ的ＰＩＡ抗原，与淋球菌ＰＩＢ无交叉反应。
Hybridoma were prepared by fusing Sp2/0 myeloma cells and spleen cells of BALB/c miceimmunized with whole-cell gonococcus.5 hybridoma cell lines were selected with purified PIA by indirect ELISA.AInong the 5 McAbs,3 were IgM class(2H11,4H8,4E10)and the other 2 were IgG1(IC2)and IgG2b(5A5)subelaases.The 2H11 and 1C2 were McAbs of high affinity. The 1C2 and5A5 recognized a same antigenic epitope of PIA. Western blotting showed that all of the 5 McAbscould specifically recognize the 35KD antigenic determinants of PIA from the complicated lysates ofwhole-cell gonococus,with no cross reaction with gonococcal PIB.
关键词
淋病奈瑟氏菌
单克隆抗体
蛋白I
亲和力
Neisseria gonorrhoeae
antibody,monoclonal
protein I
affinity
epitome
