Establishment of Anti-Dog Programmed Cell Death Ligand 1 Monoclonal Antibodies for Immunohistochemistry.

Q3 Medicine Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-02-01 Epub Date: 2024-01-18 DOI:10.1089/mab.2023.0014

Tsunenori Ouchida, Hiroyuki Suzuki, Tomohiro Tanaka, Mika K Kaneko, Yukinari Kato

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Abstract

Immune checkpoint blockade therapy has shown successful clinical outcomes in multiple human cancers. In dogs, several types of tumors resemble human tumors in many respects. Therefore, several groups have developed the anti-dog programmed cell death ligand 1 (dPD-L1) monoclonal antibodies (mAbs) and showed efficacy in several canine tumors. To examine the abundance of dPD-L1 in canine tumors, anti-dPD-L1 diagnostic mAbs for immunohistochemistry are required. In this study, we immunized the peptide in the dPD-L1 intracellular domain, and established anti-dPD-L1 mAbs, L₁Mab-352 (mouse IgG₁, kappa), and L₁Mab-354 (mouse IgG₁, kappa). In enzyme-linked immunosorbent assay, L₁Mab-352 and L₁Mab-354 showed high-binding affinity to the dPD-L1 peptide, and the dissociation constants (K_D) were determined as 6.9 × 10^-10 M and 7.2 × 10^-10 M, respectively. Furthermore, L₁Mab-352 and L₁Mab-354 were applicable for the detection of dPD-L1 in immunohistochemical analysis in paraffin-embedded dPD-L1-overexpressed cells. These results indicated that L₁Mab-352 and L₁Mab-354 are useful for detecting dPD-L1 in immunohistochemical analysis.

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建立用于免疫组化的抗犬程序性细胞死亡配体 1 单克隆抗体。

免疫检查点阻断疗法在多种人类癌症的临床治疗中取得了成功。在狗身上，有几种肿瘤在许多方面与人类肿瘤相似。因此，一些研究小组开发了抗犬程序性细胞死亡配体1（dPD-L1）单克隆抗体（mAbs），并在几种犬肿瘤中显示出疗效。要检测犬肿瘤中 dPD-L1 的含量，需要用于免疫组化的抗 dPD-L1 诊断 mAbs。在本研究中，我们对 dPD-L1 细胞内结构域的多肽进行了免疫，并建立了抗 dPD-L1 mAbs L1Mab-352（小鼠 IgG1，kappa）和 L1Mab-354（小鼠 IgG1，kappa）。在酶联免疫吸附试验中，L1Mab-352 和 L1Mab-354 与 dPD-L1 肽表现出很高的结合亲和力，其解离常数（KD）分别为 6.9 × 10-10 M 和 7.2 × 10-10 M。此外，L1Mab-352 和 L1Mab-354 还可用于石蜡包埋的 dPD-L1 表达细胞免疫组化分析中 dPD-L1 的检测。这些结果表明，L1Mab-352和L1Mab-354可用于在免疫组化分析中检测dPD-L1。

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Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Medicine-Immunology and Allergy

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4.80

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