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ADC内吞研究关键试剂——pH敏感染料标记内吞检测抗体

抗体偶联药物(ADCs)由单克隆抗体与高效细胞毒性载荷通过连接子共价连接,通过抗体实现了对癌细胞表面抗原的特异性识别与结合,从而将毒性药物精准递送至肿瘤细胞,极大降低了对正常组织的毒性。这一过程的依赖于ADC药物的核心作用机制——内吞作用。
内吞检测不仅是药物筛选的关键步骤,更是机制优化的核心手段。通过量化内化效率,科研人员能精准预测ADC在体内的行为模式,为临床试验提供关键数据支持。
为满足ADC药物内吞检测的需求,我们开发了一种抗体内吞检测试剂(Cat.No. IGG-PZF2001),该试剂利用pH敏感的荧光染料标记Fab片段,能够特异性结合待测抗体的Fc部分,10min即可形成稳定的抗体复合物,适用于快速评估抗体的内化过程,标记后抗体可用流式细胞术或细胞成像技术等多种方法检测。
Classic mechanism of action of ADC drugs

ADC的经典作用机制
(来源:https://doi.org/10.1038/s41392-022-00947-7)

产品列表

产品货号 产品描述 产品规格
IGG-PZF2001 Antibody Internalization Detection reagent 100tests/500tests

点击申请免费试用 icon

产品特征

高信噪比:阳性信号强,背景值低,信噪比高

方便快捷:仅需10min即可完成抗体标记

pH敏感:在酸性条件下发出明亮荧光,可用于胞内示踪

不影响抗原结合:靶向抗体Fc端,保留抗原结合位点

检测原理&应用场景

检测原理
抗体内化检测试剂原理图

抗体内化检测试剂原理图

应用场景
抗体发现阶段
快速筛选能特异性结合肿瘤抗原且高效内化的抗体。

药物优化阶段
评估抗体结构或偶联化学对内化过程的影响。

临床前开发阶段
确认候选药物在不同细胞模型中的内化行为。

作用机制研究阶段
验证载荷的细胞内递送过程。

验证数据

抗体内吞检测——FACS

抗体内吞检测——FACS

Anti-CD20 Abs and Human IgG1 isotype control were labeled with Antibody Internalization Detection Reagent (Cat.No.IGG-PZF2001). Raji cells were treated with Anti-CD20 Abs-Internalization Detection Reagent conjugate and Isotype control-Internalization Detection Reagent conjugate separately for 2 hours, then analysis by Flow cytometric. APC signal was used to evaluate the activity (Routine tested).

Protocol
抗体内吞检测——FACS

Anti-Her2 Abs and Human IgG1 isotype control were labeled with Antibody Internalization Detection Reagent (Cat.No.IGG-PZF2001). SK-BR-3 cells were treated with Anti-Her2 Abs-Internalization Detection Reagent conjugate and Isotype control-Internalization Detection Reagent conjugate separately for 2 hours, then analysis by Flow cytometric. APC signal was used to evaluate the activity (Routine tested).

Protocol

抗体内吞检测——荧光成像

抗体内吞检测——荧光成像

SK-BR-3 cells were treated with CellLights Lysosome GFP (green) for 16 hours followed by treatment with Anti-Her2 Abs-Internalization Detection Reagent conjugate and IgG1 Isotype-Internalization Detection Reagent conjugate separately for 16 hours (red), then stained with NucBlue Live ReadyProbes(blue) for 20 minutes and imaged on the EVOS M7000. A. Antibody Internalization Detection Reagent (Cat.No.IGG-PZF2001). B. IgG1 Isotype-Internalization Detection Reagent conjugate. C. Anti-Her2 Abs-Internalization Detection Reagent conjugate. D. Anti-Her2 Abs-Internalization Detection Reagent conjugate(Z-stacking).

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产品活动

ADC内吞检测试剂
抗Payload抗体

参考文献

  • 1. Shivatare VS, Huang HW, Tseng TH, Chuang PK, Zeng YF, Wong CH. Probing the Internalization and Efficacy of Antibody-Drug Conjugate via Site-Specific Fc-Glycan Labelling of a Homogeneous Antibody Targeting SSEA-4 Bearing Tumors. Isr J Chem. 2023 Oct;63(10-11):e202300042.

  • 2. Fu, Z., Li, S., Han, S. et al. Antibody drug conjugate: the “biological missile” for targeted cancer therapy. Sig Transduct Target Ther 7, 93 (2022).

  • 3. Nath N, Godat B, Zimprich C, Dwight SJ, Corona C, McDougall M, Urh M. Homogeneous plate based antibody internalization assay using pH sensor fluorescent dye. J Immunol Methods. 2016 Apr;431:11-21.

  • 4. Li Y, Liu PC, Shen Y, Snavely MD, Hiraga K. A Cell-Based Internalization and Degradation Assay with an Activatable Fluorescence-Quencher Probe as a Tool for Functional Antibody Screening. J Biomol Screen. 2015 Aug;20(7):869-75.

  • 背景
  • 产品列表
  • 产品特征
  • 检测原理&应用场景
  • 验证数据
  • 资源分享
  • 产品活动
  • 参考文献