Development and Characterisation of Antibody-Based Optical Imaging Probes for Inflammatory Bowel Disease

Abstract

Monoclonal antibodies are an important addition to the medicinal treatment paradigm for IBD patients. While effective, these agents show a high degree of primary and secondary non-response, and methods to predict response are highly desired. Information on drug distribution at the target level is often lacking. Fluorescent endoscopic imaging using labelled antibody drugs may provide insight regarding drug distribution, target engagement and drug response, but these assessments require stable and functional fluorescently-conjugated probes. Infliximab, vedolizumab, adalimumab and ustekinumab were conjugated to IRDye 800CW, IRDye 680LT and ZW800-1. The resulting 12 tracer candidates were analysed and characterised on SE-HPLC, SDS-PAGE, iso-electric focussing (IEF) and ELISA in order to evaluate their feasibility as candidate clinical tracers for cGMP development. Major differences in the conjugation results could be seen for each conjugated drug. For Infliximab, 2 conjugates (800CW and 680LT) showed formation of aggregates, while conjugates of all drugs with ZW800-1 showed reduced fluorescent brightness, reduced purification yield and formation of fragments. All 6 of these candidates were considered unfeasible. From the remaining 6, ustekinumab-680LT showed reduced binding to IL23, and was therefore considered unfeasible. Out of 12 potential tracer candidates, 5 were considered feasible for further development: vedolizumab-800CW, vedolizumab-680LT, adalimumab-800CW, adalimumab-680LT and ustekinumab-800CW. Infliximab-680LT and ustekinumab-680LT failed to meet the standards for this panel, but may be rendered feasible if tracer production methods were further optimized.

Orally Administrable Aggregation‐Induced Emission‐Based Bionic Probe for Imaging and Ameliorating Dextran Sulfate Sodium‐Induced Inflammatory Bowel Diseases

“…Fluorescent probes for the detection of nitric oxide (NO), [9] hypoxia, [10] caspase-1, [11] and azoreductase [12] have been rationally designed for IBD diagnosis. Furthermore, oligopeptide transporter (PepT1)-targeting fluorescent particles, [13] antibodyconjugated optical imaging probes, [14] and neutrophil formyl peptide receptor-targeting fluorescent particles [15] have been developed for IBD imaging. Nevertheless, in these fluorescent probes, cyanine dyes, coplanar donor-acceptor-donor-type molecules, squarylium dyes, and hemicyanine dyes are widely used as signal units, which are plagued by aggregation-caused emission quenching in physiological environments.…”

Detection of Tumour-Targeted IRDye800CW Tracer with Commercially Available Laparoscopic Surgical Systems

“…Bevacizumab-800CW is a tumour-targeted tracer, aimed at vascular endothelial growth factor α (VEGFα), which has been used in multiple clinical studies [9,[15][16][17][18][19][20][21][22]. Likewise, cetuximab-800CW, aimed at endothelial growth factor, is used in multiple clinical trials [23,24], and various other antibodies conjugated to IRDye800CW are currently investigated [25][26][27] since the process of developing these tracers is relatively simple compared to developing a new investigational drug or tracer [27,28]. Micro-dosages of tumour-targeted tracers are most often used in studies since first-in-patient studies can more easily be conducted due to the unlikelihood of major side effects [29][30][31][32].…”

Near-Infrared Fluorescence Imaging of EGFR-Overexpressing Tumors in the Mouse Xenograft Model Using scFv-IRDye800CW and Cetuximab-IRDye800CW

“…Fluorescence image-guided surgery, as a promising technique for intraoperative monitoring, provides real-time guidance for surgeons during surgery [4]. Fluorescent targeting agents, such as fluorescent dye-conjugated monoclonal antibodies, make it possible to image tumors with high specificity [5, 6].…”