Strategies for developing peptide and protein selective plastic antibodies

Abstract: The objective of the thesis was to explore approaches for the development of molecularly imprinted materials with predetermined selectivity for peptide or protein targets. In particular, the work has focused on molecularly imprinted nanoparticles synthesized using solid-phase synthesis protocols. Initially, previously developed glass bead-based protocols were used to prepare imprinted nanoparticles selective for neuropeptide oxytocin (Paper I), with quartz crystal microbalance (QCM) studies revealed affinity for the peptide comparable to those of commercial antibodies. In an effort to improve nanoparticle yield and to simplify aspects of nanoparticle production, a novel magnetic nanoparticle-based synthesis and separation strategy was investigated (Paper II). Using proteins pepsin and trypsin as templates, fluorescent nanoparticles were synthesized, and were shown to have picomolar affinities for their respective templates in a fluorescence-based assay format. To further develop this novel approach to imprinted nanoparticle synthesis, improvements in both the synthesis of the iron oxide core and the silica-coating of the magnetic nanoparticles were established (Paper III). Collectively, these studies have provided improved methods for the synthesis and characterization of synthetic polymer nanoparticles with antibody-like affinities for peptide and protein targets and should prove useful in the future development of plastic antibodies for diagnostic applications.