Molecularly imprinted polymers: applications in cancer diagnostics and treatment
File(s)
Author(s)
Piletsky, Stanislav
Type
Thesis
Abstract
Molecularly imprinted polymers (MIPs) are polymeric synthetic receptors that have attracted considerable attention as cost efficient and highly robust alternatives to antibodies. These polymers are generated by performing polymerisation in the presence of a template molecule, resulting in cavities that are complementary for the template. Following elution of the template, these cavities act as binding sites capable of selectively recognising the imprinted molecule.
Although MIPs were traditionally used primarily for compound separation and purification, they are now being considered as promising candidates for drug delivery and imaging. When developing MIPs for clinical applications, the choice of template molecule is essential. Additionally, the synthesis of MIPs must be optimised such that they can be produced on a large scale with excellent reproducibility. Finally these polymers must first be extensively characterised with regards to their biocompatibility. This project involved the development of a MIP-based epitope mapping technique for the identification of biomarkers suitable for targeting. This technique, dubbed ‘snapshot imprinting’, was performed on two cell lines overexpressing the cancer biomarker epidermal growth factor receptor (EGFR). This experiment identified approximately 6000 peptides from 438 proteins across the two cell lines, including 18 epitopes of EGFR.
Snapshot imprinting was then used for mapping six oesophageal cancer cell lines, identifying a further 10,500 peptides from 1363 proteins, of which many are already known to play key roles in cancer development. Finally, this technique was adapted for the mapping of tissue samples collected from ovarian cancer patients. This involved direct comparison of two tissue digestion techniques, mechanical and enzymatic digestion, yielding similar numbers and sublocations of proteins in both cases. During this work, snapshot imprinting identified a higher fraction of cell surface proteins (69%) than existing label-free deep proteomic analysis methods (27%), validating this technique for mapping of surface proteins.
Although MIPs were traditionally used primarily for compound separation and purification, they are now being considered as promising candidates for drug delivery and imaging. When developing MIPs for clinical applications, the choice of template molecule is essential. Additionally, the synthesis of MIPs must be optimised such that they can be produced on a large scale with excellent reproducibility. Finally these polymers must first be extensively characterised with regards to their biocompatibility. This project involved the development of a MIP-based epitope mapping technique for the identification of biomarkers suitable for targeting. This technique, dubbed ‘snapshot imprinting’, was performed on two cell lines overexpressing the cancer biomarker epidermal growth factor receptor (EGFR). This experiment identified approximately 6000 peptides from 438 proteins across the two cell lines, including 18 epitopes of EGFR.
Snapshot imprinting was then used for mapping six oesophageal cancer cell lines, identifying a further 10,500 peptides from 1363 proteins, of which many are already known to play key roles in cancer development. Finally, this technique was adapted for the mapping of tissue samples collected from ovarian cancer patients. This involved direct comparison of two tissue digestion techniques, mechanical and enzymatic digestion, yielding similar numbers and sublocations of proteins in both cases. During this work, snapshot imprinting identified a higher fraction of cell surface proteins (69%) than existing label-free deep proteomic analysis methods (27%), validating this technique for mapping of surface proteins.
Version
Open Access
Date Issued
2022-11-09
Date Awarded
01/02/2023
Copyright Statement
Creative Commons Attribution NonCommercial Licence
License URL
Advisor
Spivey, Alan
Aboagye, Eric
Sponsor
Engineering and Physical Sciences Research Council
Medical Research Council (Great Britain)
Imperial College London
Publisher Department
Chemistry
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)