This doctoral thesis describes the development of novel rapid deposition hydrogel platforms that serve as biointerfaces for real-time immunoassay using quartz crystal microgravimetry (QCM). Biointerface development was undertaken with the goal of developing a simple system relying on affordable technology to achieve real-time immunoassay performance equivalent to more complex and involved protocols. / The primary advantage of the hydrogel biointerfaces developed herein lies in their rapid preparation using affordable, non-toxic reagents. Compositions developed over three sequential development cycles rely on chemically cross-linking carboxymethylcellulose, which serves to covalently immobilise recognition elements through amine coupling, to polyethyleneimine. The various compositions require 10 minutes or less to deposit, a substantial improvement over competing self-assembled monolayer protocols requiring incubations ranging from hours to days using highly toxic reagents. Additional benefit lies in the immunoassay functionality of the biointerface, as these compositions excel in the traditional performance criteria of surface regeneration, minimisation of non-specific protein binding, and assay detection limit. / The peak detection limit achieved using a sandwich assay for a 17 kDa cytokine was 25 ng/mL in buffer and 500 ng/mL in a 1:3 serum dilution, with generic immunoassay capability for other cytokines demonstrated. Reusability of the developed biointerfaces is equally strong, with up to twenty regeneration cycles demonstrated without diminished sensitivity. Finally, mass-based estimates of non-specific serum adsorption indicate that the composition developed during the final design iteration equals the performance of the best protein-resistant biointerfaces currently available in the literature.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.86068 |
| Date | January 2005 |
| Creators | Carrigan, Shawn D. |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Biomedical Engineering) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 002326230, proquestno: AAINR22808, Theses scanned by UMI/ProQuest. |
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