We analyzed a comprehensive panel of commercially available CD19 detection antibody clones for IHC and FC using competition assays on tafasitamab pre-treated cell lines. To remove bound tafasitamab from the cell surface, an acidic dissociation protocol was used. Antibody affinities for CD19 were measured using Surface Plasmon Resonance (SPR) or Bio-Layer Interferometry (BLI).
While CD19 was successfully detected on tafasitamab pre-treated samples using all 7 tested IHC antibody clones, all 8 tested FC antibody clones were confirmed to compete with tafasitamab. An acidic dissociation was demonstrated essential to circumvent CD19 masking by tafasitamab and avoid false negative FC results.
The current study highlights the importance of selecting appropriate CD19 detection tools and techniques for correct interpretation of CD19 expression. The findings presented herein can serve as a guideline to investigators and may help navigate treatment strategies in the clinical setting.
我们使用在他法他单抗预处理的细胞系上的竞争测定分析了用于IHC和FC的市售CD19检测抗体克隆的综合组。要从细胞表面去除结合的他法他单抗,使用酸性解离方案。使用表面等离子体共振(SPR)或生物层干涉法(BLI)测量对CD19的抗体亲和力。
■虽然使用所有7个测试的IHC抗体克隆成功检测到他法他单抗预处理样品上的CD19,所有8个测试的FC抗体克隆均被证实与他法他单抗竞争.证明酸性解离对于规避tafasitamab的CD19掩蔽和避免FC假阴性结果是必不可少的。
■当前的研究强调了选择适当的CD19检测工具和技术以正确解释CD19表达的重要性。本文提出的发现可以作为研究者的指南,并且可以帮助在临床环境中导航治疗策略。