Site-Specific Antibody-Drug Conjugates in Triple Variable Domain Fab Format
Site-Specific Antibody-Drug Conjugates in Triple Variable Domain Fab Format
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DOI：
10.3390/biom10050764
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发表时间：
2020-05-01
期刊：
影响因子：
5.5
通讯作者：
Rader, Christoph
中科院分区：
文献类型：
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作者：
Hwang, Dobeen;Rader, Christoph
用替代抗体和抗体样支架取代单克隆抗体 (mAb) 和抗体药物偶联物 (ADC) 的传统免疫球蛋白 G (IgG) 形式的兴趣反映出需要扩大其治疗效用和效力，同时保留其精致的特异性、亲和力和低内在毒性。例如，在实体恶性肿瘤的治疗中，IgG 形式的 ADC 的肿瘤组织渗透和分布有限，减轻了细胞毒性有效负载的均匀分布。在这里，我们报告了三重可变结构域 Fab (TVD-Fab) 作为一种新格式，它可以位点特异性且稳定地生成不含 Fc 结构域的单价 ADC，且药物与抗体之比 (DAR) 为 2。TVD-Fab 具有三个可变片段 (Fv) 结构域：一个用于肿瘤靶向，两个用于通过独特的反应性赖氨酸残基快速、高效、精确和稳定地缀合两种货物。验证了 HER2 靶向 TVD-Fab 在与微管蛋白抑制剂缀合之前和之后的生化和体外细胞毒性特性。在体内，与我们之前报道的双可变域 (DVD)-IgG1 格式相比，TVD-Fab 抗体载体显示出 13.3 +/- 2.5 小时的循环半衰期和更深的肿瘤组织分布。总而言之，TVD-Fab 格式作为针对实体恶性肿瘤的位点特异性 ADC 的抗体载体值得进一步研究。
The interest in replacing the conventional immunoglobulin G (IgG) format of monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) with alternative antibody and antibody-like scaffolds reflects a need to expand their therapeutic utility and potency while retaining their exquisite specificity, affinity, and low intrinsic toxicity. For example, in the therapy of solid malignancies, the limited tumor tissue penetration and distribution of ADCs in IgG format mitigates a uniform distribution of the cytotoxic payload. Here, we report triple variable domain Fab (TVD-Fab) as a new format that affords the site-specific and stable generation of monovalent ADCs without the Fc domain and a drug-to-antibody ratio (DAR) of 2. TVD-Fabs harbor three variable fragment (Fv) domains: one for tumor targeting and two for the fast, efficient, precise, and stable conjugation of two cargos via uniquely reactive lysine residues. The biochemical and in vitro cytotoxicity properties of a HER2-targeting TVD-Fab before and after conjugation to a tubulin inhibitor were validated. In vivo, the TVD-Fab antibody carrier revealed a circulatory half-life of 13.3 +/- 2.5 h and deeper tumor tissue distribution compared to our previously reported dual variable domain (DVD)-IgG1 format. Taken together, the TVD-Fab format merits further investigations as an antibody carrier of site-specific ADCs targeting solid malignancies.