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64Cu-labeled daratumumab F(ab′)2 fragment enables early visualization of CD38-positive lymphoma

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Abstract

Purpose

Abnormal CD38 expression in some hematologic malignancies, including lymphoma, has made it a biomarker for targeted therapies. Daratumumab (Dara) is the first FDA-approved CD38-specific monoclonal antibody, enabling successfully immunoPET imaging over the past years. Radiolabeled Dara however has a long blood circulation and delayed tumor uptake which can limit its applications. The focus of this study is to develop 64Cu-labeled Dara-F(ab′)2 for the visualization of CD38 in lymphoma models.

Methods

F(ab′)2 fragment was prepared from Dara using an IdeS enzyme and purified with Protein A beads. Western blotting, flow cytometry, and surface plasmon resonance (SPR) were performed for in vitro assay. Probes were labeled with 64Cu after the chelation of 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). Small animal PET imaging and quantitative analysis were performed after injection of 64Cu-labeled Dara-F(ab′)2, IgG-F(ab′)2, and Dara for evaluation in lymphoma models.

Results

Flow cytometry and SPR assay proved the specific binding ability of Dara-F(ab′)2 and NOTA-Dara-F(ab′)2 in vitro. Radiolabeling yield of [64Cu]Cu-NOTA-Dara-F(ab′)2 was over 90% and with a specific activity of 4.0 ± 0.6 × 103 MBq/μmol (n = 5). PET imaging showed [64Cu]Cu-NOTA-Dara-F(ab′)2 had a rapid and high tumor uptake as early as 2 h (6.9 ± 1.2%ID/g) and peaked (9.5 ± 0.7%ID/g) at 12 h, whereas [64Cu]Cu-NOTA-Dara reached its tumor uptake peaked at 48 h (8.3 ± 1.4%ID/g, n = 4). In comparison, IgG-F(ab′)2 and HBL-1 control groups found no noticeable tumor uptake. [64Cu]Cu-NOTA-Dara-F(ab′)2 had significantly lower uptake in blood pool, bone, and muscle than [64Cu]Cu-NOTA-Dara and its tumor-to-blood and tumor-to-muscle ratios were significantly higher than controls.

Conclusions

[64Cu]Cu-NOTA-Dara-F(ab′)2 showed a rapid and high tumor uptake in CD38-positive lymphoma models with favorable imaging contrast, showing its promise as a potential PET imaging agent for future clinical applications.

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Funding

This work was supported by the National Natural Science Foundation of China (82171970, 81871385, 81822037, 81972446), University of Wisconsin—Madison, the National Institutes of Health (P30CA014520), the Beijing Science Foundation for Distinguished Young Scholars (JQ19028), the PKU medicine-X Youth Program (PKU2021LCXQ023), and the Open Funding Project of the State Key Laboratory of Biochemical Engineering (2020KF-01).

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Authors and Affiliations

  1. Department of Nuclear Medicine, Peking University First Hospital, Xicheng Dist, No. 8 Xishiku Str, Beijing, 100034, China

    Lei Kang, Cuicui Li, Qi Yang, Zhao Chen, Yongkang Qiu & Rongfu Wang

  2. Departments of Radiology and Medical Physics, University of Wisconsin - Madison, K6/562 Clinical Science Center, 600 Highland Ave, Madison, WI, 53705-2275, USA

    Lei Kang, Logan Sutherlin, Kaelyn V. Becker, Jonathan W. Engle, Dawei Jiang & Weibo Cai

  3. Department of Nuclear Medicine, Beijing Friendship Hospital, Beijing, 100050, China

    Cuicui Li

  4. Beijing Advanced Innovation Center for Soft Matter Science and Engineering, Beijing University of Chemical Technology, Beijing, 100029, China

    Lin Wang & Chengzhi He

  5. Department of Medical Molecular Biology, Beijing Institute of Biotechnology, 27 Tai-Ping Rd, Beijing, 100850, China

    Nan Huo & Dawei Jiang

  6. Department of Nuclear Medicine, Wuhan Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430022, China

    Xiaojie Xu

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  1. Lei Kang
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Correspondence to Lei Kang, Dawei Jiang, Xiaojie Xu or Weibo Cai.

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Weibo Cai is the scientific advisor, stockholder, and grantee of Focus-X Therapeutics, Inc. All the other authors declare no competing interests.

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Kang, L., Li, C., Yang, Q. et al. 64Cu-labeled daratumumab F(ab′)2 fragment enables early visualization of CD38-positive lymphoma. Eur J Nucl Med Mol Imaging 49, 1470–1481 (2022). https://doi.org/10.1007/s00259-021-05593-9

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