Signal loss due to oligomerization in ELISA analysis of amyloid-beta can be recovered by a novel sample pre-treatment method

被引：20
作者：
Janssen, Leen [1 ]
Sobott, Frank [2 ]
De Deyn, Peter P. [1 ,3 ,4 ,5 ,6 ,7 ]
Van Dam, Debby [1 ]
机构：
[1] Univ Antwerp, Inst Born Bunge, Lab Neurochem & Behav, Campus Drie Eiken,Univ PL 1, B-2020 Antwerp, Belgium
[2] Univ Antwerp, Dept Chem, Biomol & Analyt Mass Spectrometry & Ctr Prote CFP, B-2020 Antwerp, Belgium
[3] Hosp Network Antwerp ZNA Middelheim & Hoge Beuk, Dept Neurol, Antwerp, Belgium
[4] Hosp Network Antwerp ZNA Middelheim & Hoge Beuk, Memory Clin, Antwerp, Belgium
[5] Univ Groningen, Univ Med Ctr Groningen, Dept Neurol, Groningen, Netherlands
[6] Univ Groningen, Univ Med Ctr Groningen, Alzheimer Res Ctr, Groningen, Netherlands
[7] Univ Antwerp, Inst Born Bunge, Biobank, B-2020 Antwerp, Belgium
来源：
METHODSX | 2015年 / 2卷
关键词：
Alzheimer's disease; Amyloid-beta; ELISA; Sample pre-treatment; Oligomers; Steric hindrance;
D O I：
10.1016/j.mex.2015.02.011
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
According to the predominant theories, soluble amyloid-beta (A beta) aggregates are the principal neurotoxic agents in Alzheimer's disease pathology, making them a popular target for the development of therapeutics and diagnostic markers. One of the most commonly used methods for determining the concentration of A beta is ELISA. However, ELISA was developed for monomeric proteins and may be ill-suited for detecting aggregates. Therefore, we investigated the effect of aggregation on the ELISA measurement and developed a novel chemical pretreatment method, designed to disaggregate A beta peptides, to improve the ELISA measurement of the total A beta concentration. Synthetic A beta 40 monomers, A beta 42 oligomers and biological samples from mice and humans were subjected to a chemical pre-treatment protocol with: trifluoroacetic acid (TFA), formic acid (FA) or hexafluoroisopropanol (HFIP) prior to ELISA analysis. In our study we have shown that: A beta oligomerization leads to epitope masking and steric hindrance and results in an underestimation of the total A beta content with ELISA. Chemically pre-treating samples to disaggregate oligomers can (partially) recover the signal loss. This novel sample pre-treatment method could provide a more accurate ELISA measurement of the total A beta concentration in samples with a high oligomer content. (C) 2015 The Authors. Published by Elsevier B.V.
引用
收藏
页码：112 / 123
页数：12
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