Roadmap for the Development and Clinical Translation of Optical Tracers Cetuximab-800CW and Trastuzumab-800CW

Abstract

Optical molecular imaging using fluorescently labeled monoclonal antibodies is of significant added value in guiding surgical or endoscopic procedures. However, development of tracers for clinical trials is complex, and implementation in the clinic is therefore slow. We present a roadmap for development and translation of monoclonal antibody tracers into a drug product compliant with current good manufacturing processes (cGMPs). Methods: The production process for cetuximab-800CW and trastuzumab-800CW was optimized with regard to dye-to-protein ratio and formulation buffer. Promising formulations were produced under cGMP conditions and advanced to a full-scale stability study. Tracers were analyzed for stability by size-exclusion high-pressure liquid chromatography, pH measurement, osmolality, visual inspection, and sterility, as required by the European Pharmacopeia and cGMP guidelines. Results: Seven formulations were investigated for cetuximab-800CW and 10 for trastuzumab-800CW. On the basis of the formulation study results, we chose 2 formulations per antibody for investigation during the full-scale stability study. These formulations all performed well, showing good compliance with the acceptance criteria set for each product. Conclusion: We designed a roadmap to standardize the development, formulation, and cGMP translation of molecular fluorescent tracers. Using our standardized approach, we developed 2 stable antibody-based tracers for clinical use. The proposed roadmap can be used to efficiently develop a cGMPcompliant formulation and improve the translation of newly developed optical tracers to first-in-human use.

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

“…Application of these alternative methods could especially be beneficial for infliximab-680LT and ustekinumab-680LT, which show beneficial factors but with some problems, which may be resolvable. However, our production methods are based on methods used successfully in the past for multiple antibodies [ 33 ], and our chosen panel of analyses allow for quick assessment of several key characteristics of new antibody candidates.…”

“…Protocols for the conjugation of dye to the antibody panel were customized procedures derived from in-house labelling procedures of a clinical tracer, which was described previously [ 33 ]. Briefly, antibodies were buffer-exchanged to pH 8.5 conjugation buffer (produced in-house) using PD-10 desalting columns (Cytiva Life Sciences, Marlborough, MA, USA) and then mixed with dye in a molar dye-to-protein ratio of 2:1.…”

Fluorescent image-guided surgery in breast cancer by intravenous application of a quenched fluorescence activity-based probe for cysteine cathepsins in a syngeneic mouse model

“…To date, several biomarkers served as the targets in the development of tumor-specific probes for tumor detection in breast cancer. These markers include vascular endothelial growth factor A (VEGFA) [ 30 ], human epidermal growth factor receptor 2 (HER2) [ 31 ], gastrin-releasing peptide receptor (GRPR), and integrin α v β 3 [ 32 ]. Probes targeting these biomarkers provided high tumor-to-background ratio for tumor detection or guiding surgical resection of tumors in preclinical and early-phase clinical trials [ 21 , 28 , 32 ].…”

Improved Tumor Discrimination and Shortened Administration-to-Imaging Times in Fluorescence Guided Surgery Through Paired-Agent Protocols

“…Residual tumor increases tumor-related death at 5-years by 90% compared to those with truly negative margins (1) but the use of wide margins to remove residual tumor can lead to severe morbidity. The near ubiquitous overexpression of epidermal growth factor receptor (EGFR) -with estimates of > 90% overexpression in SCC (2,3) -has led to the development of numerous molecular therapeutic agents, which have been subsequently leveraged for fluorescence imaging (4)(5)(6)(7)(8). The goal of molecular-targeted fluorescence-guided surgery (FGS) is to improve the surgical therapeutic index based on the overexpression of the molecular target in tumor compared to normal tissue.…”