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目的利用国产IVIg初步探讨抗-Aβ的分离纯化技术。方法首先,用人工合成的人源Aβ1-42进行聚合以制备Aβ寡聚体并通过SDS-PAGE对其进行鉴定。其次,通过偶联缓冲液(0.6 mol/L柠檬酸钠、0.2 mol/L碳酸钠,PH:10)将制备好的Aβ偶联到Ultra Link Biosupport凝胶上,然后将稀释后的IVIg加入到偶联好的凝胶上,经过平衡(0.1 mol/L PBS,PH:7.4)、洗涤(0.1 mol/L PBS,PH:7.4)、洗脱(0.1 mol/L甘氨酸缓冲液,PH:2.2),从而分离纯化出抗-Aβ。最后,通过ELISA和SDS-PAGE鉴定纯化出的抗-Aβ抗体。结果聚合的Aβ1-42除了含有少量尚未聚合的单体外,主要为不同分子量的寡聚体(如:三聚体、八聚体、十二聚体等)。所获得抗-Aβ回收率为21.1%、比活为9 671.9 ng/mg、提纯倍数为248。结论使用离心柱法亲和层析可从国产IVIg制品中分离纯化出抗-Aβ。
Objective To investigate the isolation and purification of anti-Aβ using domestic IVIg. Methods First, artificial synthetic human Aβ1-42 was polymerized to prepare Aβ oligomers and identified by SDS-PAGE. Secondly, the prepared Aβ was coupled to an Ultra Link Biosupport gel by a coupling buffer (0.6 mol / L sodium citrate, 0.2 mol / L sodium carbonate, pH: 10) and then the diluted IVIg was added (0.1 mol / L PBS, pH: 7.4), washed (0.1 mol / L PBS, pH 7.4) and eluted (0.1 mol / L glycine buffer, pH: 2.2) , Thus isolated and purified anti-Aβ. Finally, the purified anti-Aβ antibody was identified by ELISA and SDS-PAGE. As a result, the aggregated Aβ1-42 mainly contains oligomers of different molecular weight (such as trimer, octamer, dodecamerand, etc.) except for a small amount of monomers that have not been polymerized. The recovery of anti-Aβ was 21.1%, the specific activity was 9 671.9 ng / mg and the purification factor was 248. Conclusion Anti-Aβ can be isolated and purified from domestic IVIg preparations by affinity column chromatography.