Amyloid-ß Oligomer Formations Over Time
Session Number
Project ID: BIO 01
Advisor(s)
Mrs. Kirsten Viola and Dr. William Klein, Northwestern University
Discipline
Biology
Start Date
17-4-2024 10:00 AM
End Date
17-4-2024 10:15 AM
Abstract
Alzheimer’s disease is progressive, fatal, and the leading cause of dementia (Chang et. al, 2003). Based on the amyloid-ß oligomer (AßOs) hypothesis, the brain damage that leads to Alzheimer’s is due to soluble, ligand-like AßOs (Lambert et. al, 1998; Cline et. al, 2018). This experiment aims to understand how time affects the formation of AßOs, specifically at near- physiological concentrations. Understanding this process will enable us to learn more about how the Aß protein changes from non-toxic monomers to neurotoxic oligomers. These experiments used Aß films to produce samples at a concentration of 1 micromolar. We collected 10 samples, flash-frozen at predetermined time-points. Western blots and SEC were used to separate and identify oligomer structures based on size. WB results suggest that initial protein conformation consists primarily of tetramers, but oligomerize into higher molecular weight structures as the tetramer becomes less abundant. Comparing SEC to WB results suggests that our antibodies are sensitive to structures that are in less abundance than what the SEC can detect and are insensitive to dimers and monomers, which show up prevalently in the SEC. All data should be repeated for confirmation, including a new calibration of the columns being used to improve confidence in results.
Amyloid-ß Oligomer Formations Over Time
Alzheimer’s disease is progressive, fatal, and the leading cause of dementia (Chang et. al, 2003). Based on the amyloid-ß oligomer (AßOs) hypothesis, the brain damage that leads to Alzheimer’s is due to soluble, ligand-like AßOs (Lambert et. al, 1998; Cline et. al, 2018). This experiment aims to understand how time affects the formation of AßOs, specifically at near- physiological concentrations. Understanding this process will enable us to learn more about how the Aß protein changes from non-toxic monomers to neurotoxic oligomers. These experiments used Aß films to produce samples at a concentration of 1 micromolar. We collected 10 samples, flash-frozen at predetermined time-points. Western blots and SEC were used to separate and identify oligomer structures based on size. WB results suggest that initial protein conformation consists primarily of tetramers, but oligomerize into higher molecular weight structures as the tetramer becomes less abundant. Comparing SEC to WB results suggests that our antibodies are sensitive to structures that are in less abundance than what the SEC can detect and are insensitive to dimers and monomers, which show up prevalently in the SEC. All data should be repeated for confirmation, including a new calibration of the columns being used to improve confidence in results.