Purpose

CD30 serves as an ideal therapeutic target for lymphoma, but its variable expression and high relapse rate pose challenges in targeted therapy. This study aims to label the anti-CD30 monoclonal antibody with ⁶⁴Cu/¹⁷⁷Lu for immuno-positron emission tomography (immuno-PET) and radioimmunotherapy (RIT).

Methods

CD30 binding kinetics of anti-CD30-IgG (IMB16) were measured by Biolayer interferometry (BLI). Western blotting screened lymphoma cell lines for CD30 expression. Flow cytometry and immunofluorescence validated the specific binding of IMB16. IMB16 was conjugated to p-SCN-Bn-NOTA(NOTA) and p-SCN-Bn-DOTA(DOTA) for radiolabeling with ⁶⁴Cu and ¹⁷⁷Lu. [⁶⁴Cu]Cu-NOTA-IMB16 and [¹⁷⁷Lu]Lu-DOTA-IMB16 were used for immuno-PET and RIT in subcutaneous lymphoma NSG mouse models.

Results

IMB16 had a strong binding affinity to CD30 according to the BLI. Western blotting revealed high CD30 expression in Karpas299 cells and negative expression in Raji cells. Flow cytometry and immunofluorescence confirmed specific binding of IMB16 to CD30 on cell surface. Radiochemical purity of [⁶⁴Cu]Cu-NOTA-IMB16 and [¹⁷⁷Lu]Lu-DOTA-IMB16 exceeded 95%. In Immuno-PET imaging, CD30-positive Karpas299 tumours had a mean uptake value of 19.2 ± 0.9%ID/g (n = 3) at 24 h post-injection, significantly higher than Karpas299-blocked and Raji-negative groups (P < 0.001). A high radiation dose (300µCi) of [¹⁷⁷Lu]Lu-DOTA-IMB16 significantly inhibited tumour growth (80.2 ± 17.6% standardized tumour volume, n = 5) at 10 days post-injection, compared to controls. Ex vivo biodistribution and histological staining supported in vivo PET imaging and RIT results.

Conclusions

Labelling IMB16 with ⁶⁴Cu enabled non-invasive assessment of CD30 expression, while ¹⁷⁷Lu labelling effectively suppressed tumour growth in CD30-positive lymphoma. CD30-targeted theranostic show promise for patient stratification and treatment enhancement, warranting further clinical evaluation.

中文翻译：

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64Cu/177Lu 标记的抗 CD30 单克隆抗体在 CD30 阳性淋巴瘤治疗诊断学中的临床前评价

 目的

CD30 是淋巴瘤的理想治疗靶点，但其可变表达和高复发率给靶向治疗带来了挑战。本研究旨在用 ⁶⁴Cu/¹⁷⁷Lu 标记抗 CD30 单克隆抗体，用于免疫正电子发射断层扫描 （immuno-PET） 和放射免疫治疗 （RIT）。

 方法

通过生物膜干涉测量法 （BLI） 测量抗 CD30-IgG （IMB16） 的 CD30 结合动力学。Western blotting 筛选淋巴瘤细胞系的 CD30 表达。流式细胞术和免疫荧光验证了 IMB16 的特异性结合。IMB16 与 p-SCN-Bn-NOTA（NOTA） 和 p-SCN-Bn-DOTA（DOTA） 偶联，用于 ⁶⁴Cu 和 ¹⁷⁷Lu 的放射性标记。[⁶⁴立方]Cu-NOTA-IMB16 和 [¹⁷⁷Lu]Lu-DOTA-IMB16 用于皮下淋巴瘤 NSG 小鼠模型的免疫 PET 和 RIT。

 结果

根据 BLI，IMB16 与 CD30 具有很强的结合亲和力。Western blotting 显示 Karpas299 细胞中 CD30 高表达，Raji 细胞中 CD30 阴性表达。流式细胞术和免疫荧光证实 IMB16 与细胞表面 CD30 的特异性结合。[⁶⁴Cu]Cu-NOTA-IMB16 和 [¹⁷⁷Lu]Lu-DOTA-IMB16 的放射化学纯度超过 95%。在免疫 PET 成像中，CD30 阳性 Karpas299 肿瘤在注射后 24 小时的平均摄取值为 19.2 ± 0.9%ID/g （n = 3），显著高于 Karpas299 阻断组和 Raji 阴性组 （P < 0.001）。与对照组相比，注射后 10 天高辐射剂量 （300μCi） [¹⁷⁷Lu]Lu-DOTA-IMB16 显着抑制肿瘤生长 （80.2 ± 17.6% 标准化肿瘤体积，n = 5）。离体生物分布和组织学染色支持体内 PET 成像和 RIT 结果。

 结论

用 ⁶⁴Cu 标记 IMB16 可以无创评估 CD30 表达，而 ¹⁷⁷Lu 标记可有效抑制 CD30 阳性淋巴瘤的肿瘤生长。CD30 靶向治疗诊断学显示出患者分层和治疗增强的前景，需要进一步的临床评估。