摘要:
用提纯的红萍鱼腥藻免疫BALB/C小白鼠的脾细胞与鼠骨髓瘤SP2/O细胞杂交融合,成功地建立了13个分泌对红萍鱼腥藻特异性单克隆抗体(McAb)的杂交瘤细胞株,杂交瘤细胞株的平均染色体数从86到105。培养液中的免疫荧光抗体(IFA)滴度为5-80,腹水中的IFA滴度为102～105。13个McAb中,11个McAb对7种红萍鱼腥藻均发生反应,1个McAb(C16)只与三膘亚属红萍鱼腥藻发生反应,而不与九膘亚属红萍鱼腥藻发生反应,具有亚属特异性,McAb-P2只识别九膘亚属中莆田羽叶萍鱼腥藻,具有种的特异性。这些McAb中,R15属于IgG1,P2属于IgG2a,C16、Me3、P1、P3、P6、R2、R6属于IgG3,P5、R1、R8、R9属于IgM,这些单克隆抗体与其他固氮蓝藻不发生反应。实验用McAb-C16和异硫氰酸荧光素(FITC)的结合物进行直接FA试验获得成功,其结合物的抗体滴度可达1280,这种反应可被特异性抗血清所阻断。
Abstract:
13 hybridoma cell lines secreting monoclonal antibody against Anabaena azollae have been established by fusion of SP2/0 myeloma cells with spleen cells from BALB/C mice immunized with Anabaena azollae. The chromosome numbers of the hybridoma cell lines range from 86 to 105, respectively. The antibodies from the culture medium have flourescencc antibody(FA)titers of up to 5-80, and those from mouse ascitic fluid have titers ranging from 102 to 105.Among the 13 monoclonal antibodies(McAbs), 11 McAbs react with all Anabaena azollae from seven species of Azolla. One(McAb-Cl6)reacts only with the symbionls from Euazolla, it did not react with that from Rhizosperma i. e., it is a subgroup-specific McAb. Another one(McAb-P2), which reacts only with the symbiont from A. pinnata, is a species-specific McAb.Two McAbs(Rl5 and P2)belong to IgG1 and IgG2a subclasses, respectively. The McAbs(Cl6, Me3,P1,P3,P6, R2 and R6)belong to IgG3 subclnss, and the remaining four McAbs(P5, R1, R8 and R9)belong to IgM subclass. All 13 McAbs did not react with free-living nitrogen fixing algae, Anabaena azotica and Tolypothrix tennis.The direct FA for Anabaena azollae was established with fluorescein isothiocyanate(FITC)labelled McAb-Cl6. The titer of McAb-Cl6 conjugated reached 1280, and the specific reaction was blocked by rabbit anti-Anabaena azollae serum.
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