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Advancing Type 1 Diabetes Screening: Evaluating the Efficacy of Improved and Optimized Assays for Enhanced Autoantibody Detection

Tipay Diaz, Jiovanni (2024) MOBN03 20232
Degree Projects in Molecular Biology
Popular Abstract
From Seedlings to Solutions: Which Test Will Sprout Success?

Type 1 diabetes (T1D) is a condition in which the body attacks its own cells that produce insulin, a necessary hormone that allows us to use energy from food. People who do not have insulin are unable to process sugar effectively, which causes major health problems. Since there is currently no cure for this condition, affected individuals require insulin on a daily basis. Researchers are working to find new ways to predict and diagnose diabetes earlier. In this project, we looked at autoantibodies, which are blood markers that can tell if a person is likely to develop diabetes. We tried new testing methods; like sowing three different seeds in the garden of innovation and see... (More)
From Seedlings to Solutions: Which Test Will Sprout Success?

Type 1 diabetes (T1D) is a condition in which the body attacks its own cells that produce insulin, a necessary hormone that allows us to use energy from food. People who do not have insulin are unable to process sugar effectively, which causes major health problems. Since there is currently no cure for this condition, affected individuals require insulin on a daily basis. Researchers are working to find new ways to predict and diagnose diabetes earlier. In this project, we looked at autoantibodies, which are blood markers that can tell if a person is likely to develop diabetes. We tried new testing methods; like sowing three different seeds in the garden of innovation and see which one would bear fruit. These seeds are called the FLuorescence ImmunoClick-IT (FLICK) to detect glutamic acid decarboxylase autoantibodies (GADA), Antibody Detection by Agglutination-PCR (ADAP) to determine the relevance of Tetraspanin 7 antibodies (TSPAN7A), and Maltose Binding Protein (MBP)-full-VAMP2 radiobinding assay to investigate the significance of vesicle-associated membrane protein 2 antibodies (VAMP2A) in T1D.

We considered adopting a novel, safer alternative to the potentially dangerous radioactive methionine used in the reference radiobinding assay. This alternative is known as L-azidohomoalanine (AHA), and we thought that it may perform the same function as radioactive methionine in labelling a protein called glutamic acid decarboxylase (GAD). Even while it seems promising, our findings suggest that it needs further improvement. Think about a lock and key system, with GAD acting as the lock and GADA serving as the key. We noticed that when we labeled GAD with AHA and processed with FLICK, the key did not fit well. This could suggest that the label made the lock a little too bulky or altered its shape somewhat, making it more difficult for the key to detect and fit into the lock. This finding is important as it emphasises the need to maintain the integrity of the lock-and-key relationship during test development.

ADAP automates the testing process using advanced technology; like having a robot in the lab doing all the repetitious labor, which speeds up results and decreases the risk of errors. Imagine this robot is assessed whether it can find a particular type of key (TSPAN7 antibody) in a large keychain. Our findings showed that the robot had difficulties to recognize TSPAN7 antibodies, and it even signaled that it found the key when it had not - these are known as "late-cycle amplification patterns" and "non-specific amplification signals." Because of these concerns, we are concluding that this robot needs some tuning to perform better.

In a study of children with T1D, we discovered that they had higher levels of antibodies against an antigen called VAMP2. The researchers employed three test conditions to specifically catch VAMP2 antibodies. The first test established a baseline, while the second and third tests attempted to fine-tune the detection, thereby confirming that the detected antibodies were actually against VAMP2. Our results suggest that VAMP2 antibodies may appear later in the disease's progression. This indicates that these antibodies could be utilized as a late-stage marker to better understand or predict how the disease will progress.

“Which Test Will Sprout Success?” It seems more correct to ask, “When will they succeed?” because each seed holds promising potential. While it is not yet their perfect season for harvest, we are optimistic. To our knowledge, no one has ever tried these techniques before, and we have now laid the groundwork for future discovery. As we continue to cultivate these seeds, we aim to harvest a future where understanding T1D with minimal laboratory risk is not just a possibility, but a practical reality. This effort moves us closer to our bigger goal, transitioning from seedlings to solutions, making life healthier and easier for individuals impacted by T1D.

Master’s Degree Project in Molecular Biology, MOBN03, 60 credits
Department of Biology, Lund University

Main Supervisor: Daniel Agardh; Co-supervisors: Alexander Lind, Samia Hamdan
Celiac and Diabetes Unit, Department of Clinical Sciences, Malmö, Faculty of Medicine, Lund University (Less)
Please use this url to cite or link to this publication:
author
Tipay Diaz, Jiovanni
supervisor
organization
course
MOBN03 20232
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
9176413
date added to LUP
2024-10-10 14:57:21
date last changed
2024-10-10 14:57:21
@misc{9176413,
  author       = {{Tipay Diaz, Jiovanni}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Advancing Type 1 Diabetes Screening: Evaluating the Efficacy of Improved and Optimized Assays for Enhanced Autoantibody Detection}},
  year         = {{2024}},
}