Institute of Blood Transfusion, Chinese Academy of Medical Sciences & Peking Union Medical College, Chengdu, China.
School of Materials Science and Engineering, Southwest Petroleum University, Chengdu, China.
Curr Alzheimer Res. 2019;16(14):1290-1299. doi: 10.2174/1567205017666200102151731.
The specific Intravenous Immunoglobulin (IVIG) for Alzheimer's Disease (AD) is developing, which contains a high level of naturally occurring autoantibodies against amyloid-β (nAbs-Aβ), and the measure of nAbs-Aβ content is greatly essential. Though Enzyme-Linked Immunosorbent Assay (ELISA) has been widely used in detecting the nAbs-Aβ content, the impact of Aβ aggregates species chosen as antigen in ELISA on this measure has not been evaluated.
To clarify the influence of different Aβ40/42 aggregates as antigen during ELISA on the content of nAbs-Aβ40/42 measured in IVIG.
Preparation of various Aβ40/42 aggregates was performed by different aggregation solutions and various lengths of time, and analyzed by western blot. Different Aβ40/42 aggregates as antigen were adopted to measure the nAbs-Aβ40/42 content in IVIG by ELISA, and the control was carried out to reduce interference of nonspecific binding. The Bonferroni and Dunnett's T3 were used for statistical analysis.
The duration for the formation of Aβ40/42 aggregates had more effect on detecting nAbs-Aβ40/42 content in IVIG than the aggregation solution. Higher content of nAbs-Aβ40/42 in the same IVIG was displayed when measured with Aβ40/42 aggregates at day 3, instead of at day 0.5 and day 7.0. The nAbs- Aβ40/42 contents in the same IVIG measured with Aβ40/42 aggregates prepared in different solutions were obviously different, but there was no significant regularity among them.
The nAbs-Aβ40/42 content in the same IVIG is significantly different when measured with Aβ40/42 aggregated under different conditions. The nAbs-Aβ40/42 content in IVIG by antigen-dependent measures, like ELISA, is uncertain.
针对阿尔茨海默病（AD）的特定静脉注射免疫球蛋白（IVIG）正在研发中，其中含有高水平的天然抗淀粉样蛋白-β（Aβ）自身抗体（nAbs-Aβ），因此，nAbs-Aβ 含量的测量至关重要。尽管酶联免疫吸附测定（ELISA）已广泛用于检测 nAbs-Aβ 含量，但 ELISA 中作为抗原的 Aβ 聚集物种类对这一测量的影响尚未得到评估。
阐明 ELISA 中使用不同的 Aβ40/42 聚集物作为抗原对 IVIG 中 nAbs-Aβ40/42 含量的影响。
采用不同的聚集溶液和不同的时间制备各种 Aβ40/42 聚集物，并通过 Western blot 进行分析。采用不同的 Aβ40/42 聚集物作为抗原，通过 ELISA 测量 IVIG 中的 nAbs-Aβ40/42 含量，并进行了对照实验以减少非特异性结合的干扰。采用 Bonferroni 和 Dunnett's T3 进行统计分析。
Aβ40/42 聚集物形成的时间对检测 IVIG 中 nAbs-Aβ40/42 含量的影响大于聚集溶液。在第 3 天测量时，同一 IVIG 中显示出更高的 nAbs-Aβ40/42 含量，而在第 0.5 天和第 7.0 天则较低。用不同溶液制备的 Aβ40/42 聚集物测量同一 IVIG 中的 nAbs-Aβ40/42 含量明显不同，但彼此之间没有明显的规律。
用不同条件下制备的 Aβ40/42 聚集物测量同一 IVIG 中的 nAbs-Aβ40/42 含量差异显著。ELISA 等基于抗原的方法测量 IVIG 中的 nAbs-Aβ40/42 含量不确定。
