Paired-Agent Imaging (PAI) is a quantitative fluorescence imaging technique that estimates the drug target concentration. It involves the co-administration of a targeted and an untargeted imaging probe to correct for nonspecific uptake and to quantify the available receptor concentration, known as the binding potential. PAI has been demonstrated in a pre-clinical setting using a 1:1 molar concentration of the targeted, ABY-029, and untargeted, IRDYE 680LT, imaging agent. However, the effects of different molar concentrations of imaging agents on the binding potential have not been studies thus far. In this study, we examined this relationship in tissue-mimicking liquid phantoms with varying molar concentration ratios. The phantom fluorescence was measured using the Pearl Imaging system and then the binding potential was quantified using MATLAB. We determined that the binding potential remains stable across concentration but increases for dye ratios where the targeted dye is higher.
Paired Agent Imaging (PAI) is a fluorescence imaging technique where a targeted probe is co-administered with an untargeted probe. PAI has been successfully demonstrated in a pre-clinical setting and its clinical translation is in progress. The tissue distribution and excretion of the two fluorescent dyes, ABY-029 and IRDY680LT, must display similar kinetics in order for the PAI model to hold. To study the excretion of the dyes, plasma studies need to be conducted to examine the presence of fluorescence in vivo over a select period of time. The current method of measuring plasma fluorescence involves centrifuging blood to isolate plasma and them measuring on a fluorometer which can be time consuming and inefficient. In this study, we examine multiple methods for visualizing and quantifying plasma fluorescence using blood and plasma phantoms at multiple concentrations. The phantom fluorescence was measured using the Pearl Imaging system and the Fluoromax-3. We have determined that imaging blood directly in a fluorescence imaging system provides the same information as plasma alone.
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