Signal loss due to oligomerization in ELISA analysis of amyloid-beta can be recovered by a novel sample pre-treatment method
According to the predominant theories, soluble amyloid-beta (Aβ) 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 concent...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-01-01
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| Series: | MethodsX |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S221501611500014X |
| _version_ | 1831601046288334848 |
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| author | Leen Janssen Frank Sobott Peter P. De Deyn Debby Van Dam |
| author_facet | Leen Janssen Frank Sobott Peter P. De Deyn Debby Van Dam |
| author_sort | Leen Janssen |
| collection | DOAJ |
| description | According to the predominant theories, soluble amyloid-beta (Aβ) 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 Aβ 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 pre-treatment method, designed to disaggregate Aβ peptides, to improve the ELISA measurement of the total Aβ concentration.
Synthetic Aβ40 monomers, Aβ42 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:
• Aβ oligomerization leads to epitope masking and steric hindrance and results in an underestimation of the total Aβ 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 Aβ concentration in samples with a high oligomer content. |
| first_indexed | 2024-12-18T14:55:42Z |
| format | Article |
| id | doaj.art-146904d2ddb84b4a99aee4d295977199 |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-12-18T14:55:42Z |
| publishDate | 2015-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-146904d2ddb84b4a99aee4d2959771992022-12-21T21:04:03ZengElsevierMethodsX2215-01612015-01-012C11212310.1016/j.mex.2015.02.011Signal loss due to oligomerization in ELISA analysis of amyloid-beta can be recovered by a novel sample pre-treatment methodLeen Janssen0Frank Sobott1Peter P. De Deyn2Debby Van Dam3Laboratory of Neurochemistry and Behavior, Institute Born-Bunge, University of Antwerp, Antwerp, BelgiumBiomolecular & Analytical Mass Spectrometry and Center for Proteomics (CFP-CeProMa), Department of Chemistry, University of Antwerp, Antwerp, BelgiumLaboratory of Neurochemistry and Behavior, Institute Born-Bunge, University of Antwerp, Antwerp, BelgiumLaboratory of Neurochemistry and Behavior, Institute Born-Bunge, University of Antwerp, Antwerp, BelgiumAccording to the predominant theories, soluble amyloid-beta (Aβ) 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 Aβ 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 pre-treatment method, designed to disaggregate Aβ peptides, to improve the ELISA measurement of the total Aβ concentration. Synthetic Aβ40 monomers, Aβ42 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: • Aβ oligomerization leads to epitope masking and steric hindrance and results in an underestimation of the total Aβ 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 Aβ concentration in samples with a high oligomer content.http://www.sciencedirect.com/science/article/pii/S221501611500014XSample pre-treatment for amyloid-beta ELISA analysis |
| spellingShingle | Leen Janssen Frank Sobott Peter P. De Deyn Debby Van Dam Signal loss due to oligomerization in ELISA analysis of amyloid-beta can be recovered by a novel sample pre-treatment method MethodsX Sample pre-treatment for amyloid-beta ELISA analysis |
| title | Signal loss due to oligomerization in ELISA analysis of amyloid-beta can be recovered by a novel sample pre-treatment method |
| title_full | Signal loss due to oligomerization in ELISA analysis of amyloid-beta can be recovered by a novel sample pre-treatment method |
| title_fullStr | Signal loss due to oligomerization in ELISA analysis of amyloid-beta can be recovered by a novel sample pre-treatment method |
| title_full_unstemmed | Signal loss due to oligomerization in ELISA analysis of amyloid-beta can be recovered by a novel sample pre-treatment method |
| title_short | Signal loss due to oligomerization in ELISA analysis of amyloid-beta can be recovered by a novel sample pre-treatment method |
| title_sort | signal loss due to oligomerization in elisa analysis of amyloid beta can be recovered by a novel sample pre treatment method |
| topic | Sample pre-treatment for amyloid-beta ELISA analysis |
| url | http://www.sciencedirect.com/science/article/pii/S221501611500014X |
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