CD30 在霍奇金淋巴瘤（HL），许多非霍奇金淋巴瘤（NHL）以及儿童和成人的非淋巴恶性肿瘤中表达。肿瘤的表达与在健康组织中的有限表达相结合，将 CD30 作为有前途的免疫疗法靶点加以鉴定。一款抗-CD30 抗体药物共轭（ADC）已获 FDA 批准用于 HL 治疗。然而，抗-CD30 ADC 和合成的抗原受体（CAR）显示了前景，但其缺点和毒性表明需要替代治疗方法。我们开发了新型的抗-CD30 x 抗-CD3 双特异性抗体（biAb），在自体患者 T 细胞（ATCs）外体激活后，涂覆于其上进行自体再输注。我们的目标是利用 biAb 的双重特异性、细胞疗法的力量以及非基因修饰自体 T 细胞输注的安全性。我们对这些 biAb 的 CD30 结合和肿瘤细胞杀伤性进行了全面描述。我们生成了对人 CD30 细胞外区域的五种独特的小鼠单克隆抗体（mAbs）。得到的抗-CD30 mAbs 被纯化并经过结合特异性、亲和力和表位识别的筛选。鉴定了两种具有独特序列和与临床使用的 ADC 不同的 CD30 结合簇的首选 mAb 候选物。这些 mAbs 与 OKT3（抗-CD3 mAb）进行化学共轭。ATCs 被武装并在体外对肿瘤细胞系进行了结合、细胞因子产生和细胞毒作用的评估，然后在体内对肿瘤细胞杀伤进行了评估。我们的首选 mAb 被亚克隆化以制备主细胞库（MCB），并对一系列人类细胞表面蛋白进行了结合筛选。只有 huCD30 被结合。这些研究支持了正在开发中的临床试验，该试验采用该新型 biAb 对自体 T 细胞进行外体负载。Copyright © 2023 Faber, Oldham, Thakur, Rademacher, Kubicka, Dlugi, Gifford, McKillop, Schloemer, Lum and Medin.

CD30 is expressed on Hodgkin lymphomas (HL), many non-Hodgkin lymphomas (NHLs), and non-lymphoid malignancies in children and adults. Tumor expression, combined with restricted expression in healthy tissues, identifies CD30 as a promising immunotherapy target. An anti-CD30 antibody-drug conjugate (ADC) has been approved by the FDA for HL. While anti-CD30 ADCs and chimeric antigen receptors (CARs) have shown promise, their shortcomings and toxicities suggest that alternative treatments are needed. We developed novel anti-CD30 x anti-CD3 bispecific antibodies (biAbs) to coat activated patient T cells (ATCs) ex vivo prior to autologous re-infusions. Our goal is to harness the dual specificity of the biAb, the power of cellular therapy, and the safety of non-genetically modified autologous T cell infusions. We present a comprehensive characterization of the CD30 binding and tumor cell killing properties of these biAbs. Five unique murine monoclonal antibodies (mAbs) were generated against the extracellular domain of human CD30. Resultant anti-CD30 mAbs were purified and screened for binding specificity, affinity, and epitope recognition. Two lead mAb candidates with unique sequences and CD30 binding clusters that differ from the ADC in clinical use were identified. These mAbs were chemically conjugated with OKT3 (an anti-CD3 mAb). ATCs were armed and evaluated in vitro for binding, cytokine production, and cytotoxicity against tumor lines and then in vivo for tumor cell killing. Our lead mAb was subcloned to make a Master Cell Bank (MCB) and screened for binding against a library of human cell surface proteins. Only huCD30 was bound. These studies support a clinical trial in development employing ex vivo-loading of autologous T cells with this novel biAb.Copyright © 2023 Faber, Oldham, Thakur, Rademacher, Kubicka, Dlugi, Gifford, McKillop, Schloemer, Lum and Medin.