Tangential flow filtration as a novel approach for fluorescent tracer purification and production up-scaling: cetuximab-800CW and ustekinumab-800CW as a proof-of-concept.

Authors

Henrik K Huizinga, Rosalie J van Dijken, Bahez Gareb, Bart G J Dekkers, Jacoba van Zanten, Hans H Westra, Pauline Koopmans, Wouter B Nagengast, Marjolijn N Lub-de Hooge

Published in

mAbs. Volume 18. Issue 1. Pages 2743444. Epub Oct 08, 2026.

Abstract

Fluorescence molecular imaging using fluorescently labeled antibodies enables identification of target cells and provides pharmacological insight into local drug distribution. The current GMP manufacturing method of tracers using PD-10 columns is subject to methodological constraints when handling low antibody concentrations, large quantities, or incompatible antibodies. This study presents Tangential Flow Filtration (TFF) as a novel and scalable approach to manufacture clinical-grade fluorescent tracers and demonstrates its applicability using cetuximab-800CW and ustekinumab-800CW. TFF system settings were optimized for purification of cetuximab-800CW and ustekinumab-800CW with labeling parameters and quality control similar to tracer production with PD-10 columns. Cetuximab-800CW production was successfully transitioned from PD-10 to TFF. QC-data after manufacturing with TFF (n = 7) or PD-10 (n = 21) were compared at release and up to 24 months of storage at 2-8°C. This revealed TFF purification to be more efficient and robust, with label efficiencies of 93.5 ± 2.3% with TFF compared to 68.3 ± 6.9% with PD-10, and 0.0 ± 0.1% unconjugated dye with TFF compared to 0.5 ± 0.3% with PD-10. Stability data were comparable for both methods. TFF manufacturing yielded clinical-grade cetuximab-800CW batches stable for at least 24 months. Concentrations ranged from 0.99-1.08 mg/mL and target binding affinity from 65-98%. Detected free dye and aggregates remained well below 3.0%. Moreover, ustekinumab-800CW was successfully manufactured using TFF after PD-10 manufacturing failed due to particle formation. Ustekinumab-800CW was stable up to 24 months at 2-8°C. Together, this demonstrates the feasibility of using TFF for clinical-grade fluorescent tracer manufacturing under GMP conditions.

PMID:
42850075
Bibliographic data and abstract were imported from PubMed on 09 Oct 2026.

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