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荧光标记蛋白

近年来，流式细胞术被广泛应用于生物药研发和生产的各个环节。荧光标记蛋白作为流式检测的首选试剂，因其预先标记了荧光素基团，可通过一步染色完成检测，既简化了检测流程、节省了检测时间，又避免了因使用二抗而产生的非特异背景。然而，高质量荧光标记蛋白的生产面临着诸多难题，如标记后蛋白活性难以维持、产品的批间一致性难以控制等，目前市场上少有高质量荧光标记蛋白可供购买。

ACROBiosystems作为专注于生物药研发领域的蛋白供应商，利用专业的蛋白研发平台、优化设计的化学标记平台及荧光定点标记平台、稳定株开发平台和流式细胞分析平台，开发了一系列高批间一致性的FITC标记和PE标记等荧光标记蛋白，适用于基于流式细胞术的分析试验，如CAR阳性表达率的检测和流式细胞术筛选抗体候选药物等。

产品特色

  1. 标记后蛋白活性经流式细胞术验证
  2. 可提供配套流式检测protocol供参考
  3. 标记前后活性保持一致
  4. 高批间一致性

应用案例

流式细胞术检测Anti-CD19 CAR阳性表达率
FACS Analysis of Anti-CD19 CAR Expression
293 cells were transfected with anti-CD19-scFv and RFP tag. 2e5 of the cells were stained with FITC-Labeled Human CD19 (20-291), Fc Tag (Cat. No. CD9-HF251, 10 μg/ml) and FITC-labeled protein control. Non-transfected 293 cells and FITC-labeled protein control were used as negative control. RFP was used to evaluate CAR (anti-CD19-scFv) expression and FITC was used to evaluate the binding activity of FITC-Labeled Human CD19 (20-291), Fc Tag (Cat. No. CD9-HF251) (QC tested).
Protocol
基于流式细胞术的配体结合试验
Flow Cytometry assay shows that PE-Labeled Human PD-1, Fc Tag, His Tag (recommended for neutralizing assay) (Cat. No. PD1-HP2F2) can bind to 293T cells overexpressing human PD-L1. 1μl stock solution per million cells (QC tested).
Protocol
基于流式细胞术的中和抗体筛选试验
FACS analysis shows that the binding of PE-Labeled Human PD-1, Fc Tag, His Tag (recommended for neutralizing assay) (Cat.No.PD1-HP2F2) to 293T overexpressing PD-L1 was inhibited by increasing concentration of neutralizing antihuman PD-L1 antibody. The IC50 is 0.29 μg/ml (QC tested).
Protocol

质控

高批间一致性
Binding activity of three different lots of FITC-labeled Human MSLN, Fc Tag was evaluated in the above FACS analysis against Anti-MSLN CAR-293 cells. The result shows very high batch-to-batch consistency.
Binding activity of three different lots of FITC-labeled Human MSLN, Fc Tag was evaluated in the above ELISA analysis against Anti-MSLN Antibody. The result shows very high batch-to-batch consistency.
标记前后活性保持一致
Binding activity of the Human MSLN, Fc Tag before and after FITC labeling was evaluated in the above ELISA analysis. The result shows that FITC-Labeled and Unconjugated Human MSLN, Fc Tag have almost the same level of binding activity.
稳定性好
Binding activity of FITC-labeled Human MSLN, Fc Tag was evaluated in the above FACS analysis against Anti-MSLN CAR-293 cells. The result shows that the FITC-labeled Human MSLN is stable at 37℃ for 24 hours without performance reduction.
Binding activity of FITC-labeled Human MSLN, Fc Tag was evaluated in the above FACS analysis against Anti-MSLN CAR-293 cells. The result shows that the FITC-labeled Human MSLN is stable within three freeze-thaw cycles without performance reduction.

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PE Labeled Protein
FITC Labeled Protein
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