Sano Masato, Kaneko Mika K, Kato Yukinari
1Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
2New Industry Creation Hatchery Center, Tohoku University, Sendai, Japan.
Monoclon Antib Immunodiagn Immunother. 2019 Aug;38(4):175-178. doi: 10.1089/mab.2019.0020. Epub 2019 Jul 29.
The diacylglycerol kinases (DGKs) catalyze the phosphorylation of the cell membrane lipid diacylglycerol (DG), which is important in lipid biochemistry and signal transduction into phosphatidic acid. DG-mediated signal transduction downstream of the T cell receptor has been reported to be terminated by DGKζ, 1 of 10 DGK isoforms in most cases. We previously established an anti-DGKζ monoclonal antibody (mAb) DzMab-1 (rat IgG, kappa), which reacts with both mouse DGKζ and human DGKζ (hDGKζ). In this study, we characterized the binding epitope of DzMab-1 using Western blotting, and found that Met1 and Pro3 residues of hDGKζ are important for facilitating DzMab-1 binding to hDGKζ. Furthermore, DzMab-1 was shown to be useful for immunohistochemical analyses for formalin-fixed paraffin-embedded HeLa cells. These findings could be applied for the production of more functional anti-hDGKζ mAbs.
二酰基甘油激酶（DGKs）催化细胞膜脂质二酰基甘油（DG）的磷酸化，这在脂质生物化学以及转化为磷脂酸的信号转导中很重要。据报道，在大多数情况下，T细胞受体下游的DG介导的信号转导由DGKζ终止，DGKζ是10种DGK同工型之一。我们之前制备了一种抗DGKζ单克隆抗体（mAb）DzMab-1（大鼠IgG，κ链），它能与小鼠DGKζ和人DGKζ（hDGKζ）发生反应。在本研究中，我们使用蛋白质印迹法对DzMab-1的结合表位进行了表征，发现hDGKζ的Met1和Pro3残基对于促进DzMab-1与hDGKζ的结合很重要。此外，DzMab-1被证明可用于对福尔马林固定石蜡包埋的HeLa细胞进行免疫组织化学分析。这些发现可应用于生产更多功能性抗hDGKζ单克隆抗体。
