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European Journal of Nuclear Medicine and Molecular Imaging
Published in: European Journal of Nuclear Medicine and Molecular Imaging 10/2021

01-09-2021 | Original Article

Radiolabelling and preclinical characterization of 89Zr-Df-radiolabelled bispecific anti-PD-L1/TGF-βRII fusion protein bintrafusp alfa

Authors: Ingrid Julienne Georgette Burvenich, Yit Wooi Goh, Nancy Guo, Hui Kong Gan, Angela Rigopoulos, Diana Cao, Zhanqi Liu, Uwe Ackermann, Christian Werner Wichmann, Alexander Franklin McDonald, Nhi Huynh, Graeme Joseph O’Keefe, Sylvia Jie Gong, Fiona Elizabeth Scott, Linghui Li, Wanping Geng, Anup Zutshi, Yan Lan, Andrew Mark Scott

Published in: European Journal of Nuclear Medicine and Molecular Imaging | Issue 10/2021

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Abstract

Purpose

Τhis study aimed to optimize the 89Zr-radiolabelling of bintrafusp alfa investigational drug product and controls, and perform the in vitro and in vivo characterization of 89Zr-Df-bintrafusp alfa and 89Zr-Df-control radioconjugates.

Methods

Bintrafusp alfa (anti-PD-L1 human IgG1 antibody fused to TGF-β receptor II (TGF-βRII), avelumab (anti-PD-L1 human IgG1 control antibody), isotype control (mutated inactive anti-PD-L1 IgG1 control antibody), and trap control (mutated inactive anti-PD-L1 human IgG1 fused to active TGF-βRII) were chelated with p-isothiocyanatobenzyl-desferrioxamine (Df). After radiolabelling with zirconium-89 (89Zr), radioconjugates were assessed for radiochemical purity, immunoreactivity, antigen binding affinity, and serum stability in vitro. In vivo biodistribution and imaging studies were performed with PET/CT to identify and quantitate 89Zr-Df-bintrafusp alfa tumour uptake in a PD-L1/TGF-β-positive murine breast cancer model (EMT-6). Specificity of 89Zr-Df-bintrafusp alfa was assessed via a combined biodistribution and imaging experiment in the presence of competing cold bintrafusp alfa (1 mg/kg).

Results

Nanomolar affinities for PD-L1 were achieved with 89Zr-Df-bintrafusp alfa and 89Zr-avelumab. Biodistribution and imaging studies in PD-L1- and TGF-β-positive EMT-6 tumour-bearing BALB/c mice demonstrated the biologic similarity of 89Zr-Df-bintrafusp alfa and 89Zr-avelumab indicating the in vivo distribution pattern of bintrafusp alfa is driven by its PD-L1 binding arm. Competition study with 1 mg of unlabelled bintrafusp alfa or avelumab co-administered with trace dose of 89Zr-labelled bintrafusp alfa demonstrated the impact of dose and specificity of PD-L1 targeting in vivo.

Conclusion

Molecular imaging of 89Zr-Df-bintrafusp alfa biodistribution was achievable and allows non-invasive quantitation of tumour uptake of 89Zr-Df-bintrafusp alfa, suitable for use in bioimaging clinical trials in cancer patients.
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Metadata
Title
Radiolabelling and preclinical characterization of 89Zr-Df-radiolabelled bispecific anti-PD-L1/TGF-βRII fusion protein bintrafusp alfa
Authors
Ingrid Julienne Georgette Burvenich
Yit Wooi Goh
Nancy Guo
Hui Kong Gan
Angela Rigopoulos
Diana Cao
Zhanqi Liu
Uwe Ackermann
Christian Werner Wichmann
Alexander Franklin McDonald
Nhi Huynh
Graeme Joseph O’Keefe
Sylvia Jie Gong
Fiona Elizabeth Scott
Linghui Li
Wanping Geng
Anup Zutshi
Yan Lan
Andrew Mark Scott
Publication date
01-09-2021
Publisher
Springer Berlin Heidelberg
Published in
European Journal of Nuclear Medicine and Molecular Imaging / Issue 10/2021
Print ISSN: 1619-7070
Electronic ISSN: 1619-7089
DOI
https://doi.org/10.1007/s00259-021-05251-0

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