目的探讨热休克蛋白70在过敏性紫癜发病机制中的作用。方法采用蛋白质免疫印迹酶联免疫吸附法(Western blot ELISA)测定37例病例组与35例对照组血浆中热休克蛋白70(HSP70)抗体。结果过敏性紫癜组中HSP70抗体检测阳性率(70.3%)明显高于...目的探讨热休克蛋白70在过敏性紫癜发病机制中的作用。方法采用蛋白质免疫印迹酶联免疫吸附法(Western blot ELISA)测定37例病例组与35例对照组血浆中热休克蛋白70(HSP70)抗体。结果过敏性紫癜组中HSP70抗体检测阳性率(70.3%)明显高于对照组(17.1%),两者相比差异有显著性(P<0.01);过敏性紫癜肾炎型HSP70抗体阳性率(80%)高于非肾炎型(63.6%),两者相比差异有显著性(P<0.05)。结论HSP70参与了过敏性紫癜的发病过程,且可反应肾脏是否受累。展开更多
Recognizing proteins via the production of highly specific monoclonal antibodies(mAbs)is crucial to identifying proteins for proteomic research.However,traditional mAb generation is time-consuming with low efficiency....Recognizing proteins via the production of highly specific monoclonal antibodies(mAbs)is crucial to identifying proteins for proteomic research.However,traditional mAb generation is time-consuming with low efficiency.In this study,we assessed the high throughput method of producing mAbs by immunizing mice with multiple antigens in order to obtain hybridomas against these multiple antigens in one cell fusion.We selected eight proteins that play important roles in human physiological or pathological processes.These proteins were mixed and simultaneously administered to one mouse.We observed the immunizing period for 10 d,and determined the effect of liquid medium and semi-solid medium in hybridoma generation.As a result,all eight immunogens induced antibodies in the immunized mouse in one cell fusion,we obtained hybridomas specific to all eight proteins by enzyme-linked immuno sorbent assay(ELISA)screening,hybridomas against five out of eight showed specific positive in Western-blotting assays.This indicates that we generated mAbs specific to eight proteins in one cell fusion,greatly increasing the efficiency of mAb generation.Furthermore,we observed that hybridomas selected from the liquid medium and semi-solid medium showed different reactivity to antigens.Our study established high-throughput and time-saving methods for production of mAbs.These results provide alternative approaches for increasing the efficacy of mAb generation.展开更多
文摘目的探讨热休克蛋白70在过敏性紫癜发病机制中的作用。方法采用蛋白质免疫印迹酶联免疫吸附法(Western blot ELISA)测定37例病例组与35例对照组血浆中热休克蛋白70(HSP70)抗体。结果过敏性紫癜组中HSP70抗体检测阳性率(70.3%)明显高于对照组(17.1%),两者相比差异有显著性(P<0.01);过敏性紫癜肾炎型HSP70抗体阳性率(80%)高于非肾炎型(63.6%),两者相比差异有显著性(P<0.05)。结论HSP70参与了过敏性紫癜的发病过程,且可反应肾脏是否受累。
基金supported by the National Basic Research Program of China (2011CB915502)State Key Laboratory of Proteomics Project (SKLP Y200907)National High Technology Research and Development Program of China (2012AA020201)
文摘Recognizing proteins via the production of highly specific monoclonal antibodies(mAbs)is crucial to identifying proteins for proteomic research.However,traditional mAb generation is time-consuming with low efficiency.In this study,we assessed the high throughput method of producing mAbs by immunizing mice with multiple antigens in order to obtain hybridomas against these multiple antigens in one cell fusion.We selected eight proteins that play important roles in human physiological or pathological processes.These proteins were mixed and simultaneously administered to one mouse.We observed the immunizing period for 10 d,and determined the effect of liquid medium and semi-solid medium in hybridoma generation.As a result,all eight immunogens induced antibodies in the immunized mouse in one cell fusion,we obtained hybridomas specific to all eight proteins by enzyme-linked immuno sorbent assay(ELISA)screening,hybridomas against five out of eight showed specific positive in Western-blotting assays.This indicates that we generated mAbs specific to eight proteins in one cell fusion,greatly increasing the efficiency of mAb generation.Furthermore,we observed that hybridomas selected from the liquid medium and semi-solid medium showed different reactivity to antigens.Our study established high-throughput and time-saving methods for production of mAbs.These results provide alternative approaches for increasing the efficacy of mAb generation.
