Development of an ELISA to accurately measure the solubility of the amyloid beta 42 molecule
(2024) KFKM05 20241Biophysical Chemistry
- Abstract (Swedish)
- Amyloid beta (Aβ) proteins are a group of proteins able to form fibrillar aggregates that are linked to the adverse neurological effects seen in Alzheimer’s disease [11]. The molecular mechanisms that lead to the Aβ aggregation are complex processes, and efforts to understand them are being made by investigating the kinetics and thermodynamic properties of the protein. The solubility of the protein is a central thermodynamic property governing the equilibrium between the non-aggregated forms and the aggregated forms [10]. To study the solubility, it is necessary to be able to study both aggregated and non-aggregated states. One method to study the non-aggregated state is with an enzyme-linked immunosorbent assay (ELISA). The aim of this... (More)
- Amyloid beta (Aβ) proteins are a group of proteins able to form fibrillar aggregates that are linked to the adverse neurological effects seen in Alzheimer’s disease [11]. The molecular mechanisms that lead to the Aβ aggregation are complex processes, and efforts to understand them are being made by investigating the kinetics and thermodynamic properties of the protein. The solubility of the protein is a central thermodynamic property governing the equilibrium between the non-aggregated forms and the aggregated forms [10]. To study the solubility, it is necessary to be able to study both aggregated and non-aggregated states. One method to study the non-aggregated state is with an enzyme-linked immunosorbent assay (ELISA). The aim of this project was to develop an ELISA for the monomeric form of the Aβ42 peptide. The method was based on a previous ELISA described in Hellstrand et al.(2009) [8]. The goals of the method development was for the ELISA to be sensitive and to be selective for the Aβ42 monomer. With the method, an Aβ42 aggregation kinetics experiment described in Hellstrand et al. (2009) was reproduced. The method was developed for Aβ42 in a buffer environment, but the assay was also tested with Aβ42 in an environment of cerebrospinal fluid (CSF).
The method development was successful and the method showed satisfactory sensitivity and selectivity. Standard curves with linear regions between 1 and ∼30 nM Aβ42 were obtained. The limit of detection and limit of quantification of the method were shown to be in the low single-digit nanomolar range. The results from the aggregation kinetics experiment and the experiment described in the Hellstrand et al. (2009) paper varied slightly, possibly because of the ELISA being saturated or because of different fibril formations being achieved. The CSF experiments showed that unprocessed CSF has an interfering effect on the assay. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9167772
- author
- Ekström, Elsa LU
- supervisor
-
- Sara Linse LU
- Max Lindberg LU
- Kalyani Sanagavarapu LU
- organization
- course
- KFKM05 20241
- year
- 2024
- type
- H3 - Professional qualifications (4 Years - )
- subject
- keywords
- amyloid beta, amyloid beta 42, alzheimer's disease, ELISA, enzyme-linked immunosorbent assay, amyloid beta 42 solubility, amyloid beta aggregation, biophysical chemistry
- language
- English
- id
- 9167772
- date added to LUP
- 2024-08-08 13:23:23
- date last changed
- 2024-08-08 13:23:23
@misc{9167772,
abstract = {{Amyloid beta (Aβ) proteins are a group of proteins able to form fibrillar aggregates that are linked to the adverse neurological effects seen in Alzheimer’s disease [11]. The molecular mechanisms that lead to the Aβ aggregation are complex processes, and efforts to understand them are being made by investigating the kinetics and thermodynamic properties of the protein. The solubility of the protein is a central thermodynamic property governing the equilibrium between the non-aggregated forms and the aggregated forms [10]. To study the solubility, it is necessary to be able to study both aggregated and non-aggregated states. One method to study the non-aggregated state is with an enzyme-linked immunosorbent assay (ELISA). The aim of this project was to develop an ELISA for the monomeric form of the Aβ42 peptide. The method was based on a previous ELISA described in Hellstrand et al.(2009) [8]. The goals of the method development was for the ELISA to be sensitive and to be selective for the Aβ42 monomer. With the method, an Aβ42 aggregation kinetics experiment described in Hellstrand et al. (2009) was reproduced. The method was developed for Aβ42 in a buffer environment, but the assay was also tested with Aβ42 in an environment of cerebrospinal fluid (CSF).
The method development was successful and the method showed satisfactory sensitivity and selectivity. Standard curves with linear regions between 1 and ∼30 nM Aβ42 were obtained. The limit of detection and limit of quantification of the method were shown to be in the low single-digit nanomolar range. The results from the aggregation kinetics experiment and the experiment described in the Hellstrand et al. (2009) paper varied slightly, possibly because of the ELISA being saturated or because of different fibril formations being achieved. The CSF experiments showed that unprocessed CSF has an interfering effect on the assay.}},
author = {{Ekström, Elsa}},
language = {{eng}},
note = {{Student Paper}},
title = {{Development of an ELISA to accurately measure the solubility of the amyloid beta 42 molecule}},
year = {{2024}},
}