摘要
目的建立一种检测帕金森病(Parkinson’s disease,PD)患者血浆促进外源性α-突触核蛋白(α-synuclein,α-Syn)寡聚体形成的方法。方法利用本室自制的鼠抗人α-Syn单克隆抗体建立检测α-Syn寡聚体的酶联免疫吸附测定(enzyme linked immunosorbent,ELISA)方法。将不同质量浓度(200、100、50、25、12.5μmol/L)重组人α-Syn在正常人和PD患者血浆中分别振荡孵育(37℃、280 r/min)24、48、72、96 h。用所建立的ELISA法检测振荡后血浆中的α-Syn寡聚体含量。结果所建立ELISA方法可以特异性检测α-Syn寡聚体,不识别α-Syn单体。重组人α-Syn在质量浓度为100μmol/L、血浆稀释3倍以下、振荡孵育48h条件下可以在PD患者血浆中形成较多的寡聚体,并与对照血浆中形成的α-Syn寡聚体含量对比差异最大。结论成功建立了一种检测PD患者血浆促进α-Syn寡聚体形成能力的方法,该方法可以用于诊断PD患者血浆的潜在病理变化。
Objective To establish a method for evaluation of exogenous α-synuclein （α-Syn） oligomer formation in plasma of patients with Parkinson＇ s disease（PD）. Methods An ELISA method was developed by using the 3D5 mouse anti-human a-Syn monoclonal antibody. Various concentrations of α-Syn（200, 100, 50, 25, 12.5 μmol/L） was incubated（37℃ , 280 r/min） in PD and control sera for 24, 48, 72, and 96 h. The amounts of α-Syn oligomers formed in the sera were measured using the established ELISA method. Results The ELISA method established specifically detected α-Syn oligmers with no reaction to α-Syn monomers. The best differentiation between the α-Syn oligomer formation in PD and control sera was observed when the recombinant α-Syn concentration was 100 μmol/L, with the incubation time being 48 h and the plasma diluted to less than 1/3. Conclusion A method for evaluation of α-Syn oligomer formation in the sera of PD patients and normal controls was established, which can differentiate the potential pathological changes of PD.
出处
《首都医科大学学报》
CAS
2013年第6期830-834,共5页
Journal of Capital Medical University
基金
国家自然科学基金(81071014)
国家重点基础研究发展计划(2011CB504101)
首都卫生发展科研专项课题(首发2011-1001-01)
国家科技支撑计划课题(2012BAI10B03)
北京市自然科学基金(7102076)~~
