【作者】 米友军； 苏露； 刘海； 周建国； 傅思武；

【机构】 西北民族大学医学院；

【摘要】 【目的】:制备抗艰难梭菌B毒素的单克隆抗体（mAb）并鉴定其特性。【方法】:用纯化的艰难梭菌B毒素免疫BALB/c小鼠,将免疫小鼠的脾细胞与骨髓瘤细胞Sp2/0融合,采用间接ELISA筛选杂交瘤细胞。用ELISA检测mAb腹水的效价;用Western blot检测mAb的特异性。【结果】:得到1株杂交瘤细胞株（384）,该株细胞分泌的mAb为IgG2a。中和试验表明,该mAb均无中和活性。腹水mAb的效价均在10^-4以上。在非变性条件下,PAGE后Western blot的结果显示,该株mAb可与相对分子质量（Mr）为22×10⁴的B毒素产生反应;而在变性条件下,SDS-PAGE后Western blot均显示,可与Mr为4×10³的B毒素片段产生反应。【结论】:该株杂交瘤细胞株能分泌抗艰难梭菌B毒素的特异性mAb,为艰难梭菌B毒素的研究提供了有利的工具。更多还原

【Abstract】 Aim:To prepare monoclonal antibody(mAb) against Clostridium difficile toxin B and identify its properties.method:BALB/c mice were immunized with C.difficile toxin B.The splenocytes from immunized mice were fused with myeloma cells Sp2/0.The hybridoma cells were screened by indirect ELISA and limiting dilution method.The titer of ascitics of mAb was determined by ELISA.Specificity of mAb was analyzed by Western blot. Result:One hybridoma cell line(3B4) secreting mAb against C.difficile toxin B was obtained.The Ig subclass of mAb 3B4 was IgG2a.mAb had no neutralization activity. Western blot analysis Native-PAGE shows that mAb reacted to C.difficile toxin B with M_r being 22×10~4,and under the condition of SDS-PAGE,Western blot analysis showed that mAb reacted to subunits of C.difficile toxin B with M_r being 4×10~3. Conclusion:mAb against C.difficile toxin B with high titers was obtained successfully with satisfactory specificity,which will be useful for detection and research of C.difficile toxin B更多还原