Effect of nonspecific binding of imaging agents to plasma protein in the paired-agent imaging for resection during surgery (PAIRS)

Long-term survival of head and neck squamous cell carcinoma (HNSCC) patients have proven to be correlated with negative surgical margins. Paired-Agent Imaging for Resection during Surgery (PAIRS) is capable of drawing the fine line between tumor and normal tissue by employing a control imaging-agent, which is co-administered with the targeted imaging agent to account for nonspecific signal. PAI is highly dependent on the parallel paired-agent delivery and static quantum yield of the agent to trace the molecular concentration. However, it is well known that nonspecific binding of fluorescence probes to plasma proteins can change its delivery, dissociation constant, and quantum yield. A thorough evaluation of the effect of plasma protein binding in the estimation of receptor concentration was performed for the paired-agents in this study. We are planning to evaluate ABY-029, an anti-epithelial growth factor receptor (EGFR) Affibody, and IRDye 700DX as a control agent. The plasma-dependent change in fluorescence intensity, percent binding, and in vivo distribution kinetics will be studied for each agent alone, and in combination. In this proceeding, the absorption, emission patterns for the targeted agent, ABY-029, measured by UV-Vis, fluorometer, and Pearl were shown. Initial studies indicate that binding to Bovine serum albumin (BSA), human serum albumin (HSA) and EGFR can introduce the Solvatochromic shift, which will change the absorption and emission pattern for ABY-029. Computational modeling will be performed to determine how each of these changes will affect the determined BP, and thus detection of tumors from normal tissue.