摘要
目的制备稳定分泌Aβ2 2 -35单克隆抗体的杂交瘤细胞 ,以Aβ2 2 -35单克隆抗体建立Aβ检测方法。 方法以人工合成的Aβ2 2 -35连接匙孔槭血蓝蛋白 ,免疫Balb/c小鼠 ,取脾细胞与SP2 / 0 细胞融合 ,制得能稳定分泌Aβ2 2 -35单克隆抗体的杂交瘤细胞 ,采用Westernblotting增强化学发光技术观察大鼠脑内Aβ含量。 结果得到 2株能稳定分泌Aβ2 2 -35单克隆抗体的杂交瘤细胞 ,Aβ2 2 -35单克隆抗体的Ig亚类为IgG3 ,青年组和老年组大鼠脑Aβ含量分别为 9.8± 2 .8和 13 .3 6± 2 .65 pmol/ 12mg脑组织。 结论制备得到的Aβ2 2 -35单克隆抗体特异性强 ,效价高 ,测得的老年大鼠脑Aβ含量显著高于青年大鼠。
Objective To produce a hybridoma secreting stable monoclonal antibodies against Aβ 22-35 and to develop a detection method for the assay of Aβ. Spleen cells from Balb/c mice immunized with Aβ 22-35 -KLH were fused with mouse myeloma cells SP 2/0 . The techniques of immunoprecipitation and western blotting plus ECL were used to investigate the levels of Aβ in the rat brain. Two strains of hybridomas (3A 8 and 3B 2) secreting stable monoclonal antibodies raised against Aβ 22-35 were obtained. The subtypes of Aβ 22-35 were IgG 3. The levels of Aβ in young and older rat brain were 9.8±2.8 and 13.36±2.65 (pmol/12mg brain tissues, x±s), respectively. The Aβ 22-35 mAb obtained had high titres and specificity. The levels of Aβ in the older rat brain were significantly increased as compared with the young one (P<0.05).
出处
《上海第二医科大学学报》
CSCD
2001年第2期110-112,140,共4页
Acta Universitatis Medicinalis Secondae Shanghai
基金
国家自然科学基金资助课题 (39570 869)
