Fluorescent bioimaging has proven to be a powerful tool in non-invasively studying biological processes in living systems. One application of this technique is being used in a transgenic zebrafish model to study the innate immune response to infection of Candida Albicans. Methods utilizing genetic engineering, however, are limited by time constraints that arise when having to depend upon the ability to modify genetics of the model. To bypass these constraints, this project seeks to produce a modified version of a previously developed biofluorescent probe, which was validated in a mouse model. The probe consists of three components, a hexapeptide, a fluorophore, and a polyethylene glycol (PEG) polymer. The peptide was found to bind to the formyl peptide receptors (FPRs) of the neutrophils in mice. Synthesis of this peptide was accomplished using solid phase peptide synthesis and standard Fmoc-chemistry. The peptide was purified via high performance liquid chromatography (HPLC), and characterized via mass spectrometry and 1H NMR and 13C NMR. The product had a yield of 9 mg. The PEG polymer serves to increase bioavailability and was conjugated to the peptide in sodium borate acetonitrile buffer system. This molecule was characterized via MALDI mass spectrometry and resulted in a yield of 4.9 mg. Further studies will verify the successful conjugation of the fluorophore to the PEG polymer. Once characterized, the probe will be validated through application to an unmodified zebrafish model.
Novak, Sadie Xiaohua, "Synthesis of a Fluorescently Tagged Bioactive Probe Specific to Neutrophils in a Zebrafish Model" (2019). Honors College. 560.