Thesis

Selection and characterization of DNA aptamers for therapeutic monitoring of Lenalidomide, 6-Mercaptopurine, Dabrafenib and Venetoclax

Downloadable Content

Download PDF
Creator
Rights statement
Awarding institution
  • University of Strathclyde
Date of award
  • 2026
Thesis identifier
  • T17578
Person Identifier (Local)
  • 201879829
Qualification Level
Qualification Name
Department, School or Faculty
Abstract
  • Adverse drug reactions associated with some anticancer medicines are a significant reason for patient compliance; therefore, healthcare providers have invested substantial effort in the last four decades in therapeutic drug monitoring (TDM) research to maximize therapeutic results, minimize toxicity or to achieve both of these. Therapeutic drug monitoring (TDM) is extremely important for anticancer drugs which often have serious adverse effects and whose optimum dosage can differ significantly between patients. Most analytical techniques applied for TDM are chromatographic methods, such as HPLC, LC-MS and LC-MSMS. Since these current methods have some challenges, such as time consuming, financial cost, and the need for high skilled personnel, it was beneficial to search for better methods. Therefore, the aim of this research to study the developing and synthesizing of an aptamer model to approach monitoring of Lenalidomide (LDM), 6-Mercaptopurine (6-MP), Dabrafenib (DFB) and Venetoclax (VENX) via the Sequential Evolution of Ligands by Exponential Enrichment (SELEX) process. In this work, DNA aptamers against four commonly used anticancer – Lenalidomide (LDM), 6-Mercaptopurine (6-MP), Dabrafenib (DFB) and Venetoclax (VENX) – have been selected, identified and characterized through the systematic evolution of ligands by exponential enrichment (SELEX) technique. Ten cycles of selection have been applied for each medicine. Many different Lenalidomide, 6-Mercaptopurine, Dabrafenib and Venetoclax-specific aptamer sequences were successfully obtained. For the Lenalidomide, 19 aptamers were isolated, identified and aligned through SELEX and cloning then three of these were synthesized and the binding affinity of these three candidate’s ssDNA aptamers to Lenalidomide were individually tested by a microscale thermophoresis (MST) technique and their dissociation constants calculated (Kd for the candidate aptamer number 1 = 653.46 ± 0.23 nM; Kd for the candidate aptamer number 2 = 15.18 ± 0.37 nM; and Kd for the candidate aptamer number 3 = 7.75 ± 0.09 nM). For the 6-Mercaptopurine, 10 aptamers were isolated, identified and aligned through SELEX and cloning and then three of these were synthesized. The binding affinity of these three candidate’s ssDNA aptamers to 6- Mercaptopurine were individually tested by microscale thermophoresis (MST) and their dissociation constants calculated (Kd for the candidate aptamer number 1 = 2.30 ± 0.22 nM; Kd for the candidate aptamer number 2 = 46.8 ± 0.23 nM; and Kd for the candidate aptamer number 3 = 0.1552 ± 0.37 nM). For Dabrafenib, 28 aptamers were isolated, identified and aligned through SELEX and cloning, then three of these were synthesized and the binding affinity of these three candidate’s ssDNA aptamers to Dabrafenib were individually tested by microscale thermophoresis (MST) with their dissociation constants calculated (Kd for the candidate aptamer number 1 = 0.324 ± 0.23 nM; Kd for the candidate aptamer number 2 = 0.153 ± 0.23 nM; and Kd for the candidate aptamer number 3 = 2.016 ± 0.23 uM). Finally, for the Venetoclax, eight aptamers were isolated, identified and aligned through SELEX and cloning. Three of these were synthesized and the binding affinity of these three candidate’s ssDNA aptamers to Venetoclax were individually tested by microscale thermophoresis (MST). Their dissociation constants calculated (Kd for the candidate aptamer number 1 = 29.06 ± 0.28 nM; Kd for the candidate aptamer number 2 = 11.66 ± 0.24 nM; and Kd for the candidate aptamer number 3 = 13.87 ± 0.29 uM). This work reports the first oligonucleotide aptamers selected for Lenalidomide, 6-Mercaptopurine, Dabrafenib and Venetoclax with Kd in the nanomolar range. These aptamers can be used for both basic research and clinical purposes. The sub-nanomolar range of Kds indicate that these aptamers have very high affinity for their respective drugs. These aptamers could be an emerging molecular recognition receptor for the construction of highly specific and very sensitive aptamer-based biosensors for therapeutic drug monitoring applications.
Advisor / supervisor
  • Khadra, Ibrahim
Resource Type
Note
  • The restriced access statement on each page of this thesis DOES NOT apply.
DOI
Alternative Title
  • Selection and characterisation of DNA aptamers for therapeutic monitoring of Lenalidomide, 6-Mercaptopurine, Dabrafenib and Venetoclax
Date Created
  • 2025

Relations

Items

Thumbnail Title Date Uploaded Visibility Actions
file details PDF of thesis T17578 2026-02-04 Public Download