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>技术资源 > 前沿速递 >【技术干货】基于MOA的报告基因生物活性检测方法

【技术干货】基于MOA的报告基因生物活性检测方法

2022-04-08 09:38    浏览量：3467




活性测定是对药物的有效成分和含量以及药物效价的测定，是确保抗体类药物有效性的重要质控指标。报告基因细胞株（Reporter cell line）以药物作用机制（MOA）为基础，通过基因工程方法构建，在细胞内建立起靶点转导信号与报告基因表达的相关性，通过检测报告基因的表达情况以监控靶点信号通路转导，进而构建一种药物靶点特异性的模型细胞株。这类细胞株批间差小，且转入细胞内的检测信号强，以此为基础开发的分析方法灵敏度高，检测周期短，重复性好，并且操作简单，因此被认为可以克服传统细胞水平分析方法劣势而逐步被广泛运用到新药研发的进程中，并在2020 年版的《中国药典》中被收录为血管内皮生长因子(VEGF)抗体及I型干扰素(IFN)活性测定的标准方法之一。




为满足新药物研发的需求，ACROBiosystems采用荧光素酶报告基因系统(Luciferase Reporter Assay System)建立报告基因细胞平台，持续开发系列高质量报告基因细胞株产品。ACRO开发的系列细胞株产品均通过功能性和稳定性验证，可以应用于信号传导功能研究，早期药物发现，高通量药物筛选、生物活性测定、稳定性测定、批次放行等方面，为新药研发提供一个稳定且便捷的工具。

产品优势

  • 基于明确MOA设计，便于药物机制研究

  • 反应信号强，确保高灵敏度

  • 检测窗口大， 适用不同药效筛选

  • 代次稳定性好，利于方法学验证

  • 应用场景明确，方便客户选择

  • 配套产品供应，实现一站服务<

应用场景

  • 细胞内信号转导通路研究

  • 配体受体之间的相互作用研究

  • 早期药物筛选和新药研发

热门产品
货号 Host cell 产品描述 Applications 订购/预购
CHEK-ATF044 HEK293 VEGFR2 (Luc) HEK293 Reporter Cell Screen for anti-human VEGF or anti-human VEGFR neutralizing antibody

订购

CHEK-ATF045 HEK293 TSLPR (Luc) HEK293 Reporter Cell Screen for anti-human TSLP or anti-human TSLPR neutralizing antibody

订购

CJUR-STF046 Jurkat NFAT (Luc) Jurkat Reporter Cell Screen for T cell activators

订购

CHEK-ATF047 HEK293 STAT3 (Luc) HEK293 Reporter Cell Screen for STAT3 activators or inhibitors

订购

CHEK-ATF048 HEK293 NF-κB (Luc) HEK293 Reporter Cell Screen for NF-kB inhibitors and activators.

订购

CHEK-ATF049 HEK293 EGFR (Luc) HEK293 Reporter Cell Screen for anti-human EGFR or anti-human EGF neutralizing antibody or small molecule inhibitor

订购

CHEK-ATF050 HEK293 NFAT (Luc) HEK293 Reporter Cell Screen NFAT activators or inhibitors

订购

>>>如果您希望ACRO开发更多类型的报告基因细胞系产品，请点击提议


验证数据&应用案例

1. TSLPR (Luc) HEK293 Reporter Cell (Cat. No. CHEK-ATF045)

  • 细胞性质验证

经FACS及Signaling Bioassay验证，Cat. No. CHEK-ATF045表达TSLPR及IL7R，用连续稀释的TSLP蛋白 (Cat. No.TSP-H52Hb)进行刺激，EC50值为0.1392 μg/mL，最大诱导倍数约为 45.15。

Co-expression analysis of human TSLPR and IL7Rα on TSLPR (Luc) HEK293 Reporter Cell by FACS. Cell surface staining was performed on TSLPR (Luc) HEK293 Reporter Cell or negative control cell using PE-labeled anti-TSLPR antibody and FITC-labeled anti-IL7Rα antibody.

点击申请Protocol

Response to human TSLP protein (RLU). The TSLPR (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human TSLP protein (ACROBiosystems, Cat.No.TSP-H52Hb). The EC50 was approximately 0.1392 μg/mL.

点击申请Protocol

Response to human TSLP protein (Fold). The TSLPR (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human TSLP protein (ACROBiosystems, Cat.No.TSP-H52Hb). The max induction fold was approximately 45.15.

点击申请Protocol


Passage stability analysis of receptors expression by FACS. Flow cytometry surface staining of human TSLPR and IL7Rα on TSLPR (Luc) HEK293 Reporter Cell demonstrates consistent mean fluorescent intensity across across passage10-26. (A) Human IL7Rα expression analysis. (B) Human TSLPR expression analysis.

点击申请Protocol


Passage stability analysis by Signaling Bioassay. The continuously growing TSLPR (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human TSLP protein. Human TSLP protein stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 10-26.

点击申请Protocol


  • 应用案例

经验证，抗TSLP抗体可以抑制TSLP蛋白(Cat. No. TSP-H52Hb)诱导的报告基因活性，EC50值为0.55 μg/mL。

Inhibition of human TSLP protein-induced reporter activity by anti-human TSLP neutralizing antibody. This reporter cell was incubated with serial dilutions of antibodies in the presence of human TSLP protein (ACROBiosystems, Cat. No.TSP-H52Hb) with a final concentration of 0.3 μg/mL. 

点击申请Protocol



2. EGFR (Luc) HEK293 Reporter Cell (Cat. No. CHEK-ATF049)

  • 细胞性质验证

经FACS及Signaling Bioassay验证，Cat. No. CHEK-ATF049表达EGFR，用连续稀释的EGF蛋白 (Cat. No. EGF-H52H3)进行刺激，EC50值为56.23 ng/mL，最大诱导倍数约为56。

Expression analysis of human EGFR on EGFR (Luc) HEK293 Reporter Cell by FACS. Cell surface staining was performed on EGFR (Luc) HEK293 Reporter Cell or negative control cell using PE-labeled anti-EGFR antibody.

点击申请Protocol

Response to human EGF protein (RLU). The EGFR (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human EGF protein (ACROBiosystems, Cat.No.EGF-H52H3). The EC50 was approximately 56.23 ng/mL.

点击申请Protocol

Response to human EGF protein (Fold). The EGFR (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human EGF protein (ACROBiosystems, Cat.No.EGF-H52H3). The max induction fold was approximately 56.

点击申请Protocol


  • 代次稳定性验证

通过FACS及Signaling Bioassay对Cat. No. CHEK-ATF049传代稳定性进行验证，均证明该细胞株可稳定传递10-20代。

Passage stability analysis of receptor expression by FACS. Flow cytometry surface staining of human EGFR on EGFR (Luc) HEK293 Reporter Cell demonstrates consistent mean fluorescent intensity across across passage10-20.

点击申请Protocol

Passage stability analysis by Signaling Bioassay. The continuously growing EGFR (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human EGF protein. Human EGF protein stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 10-20.

点击申请Protocol


  • 应用案例

筛选中和抗体：经验证，抗EGFR抗体可以抑制EGF蛋白 (Cat. No. EGF-H52H3)诱导的报告基因活性，EC50值为1.793 μg/mL。

Inhibition of human EGF protein-induced reporter activity by anti-human EGFR neutralizing antibody. This reporter cell was incubated with serial dilutions of antibodies in the presence of human EGF protein (ACROBiosystems, Cat. No. EGF-H52H3) with a final concentration of 50 ng/mL. The EC50 of anti-human EGFR neutralizing antibody (Cetuximab) is approximately 1.793 μg/mL.

点击申请Protocol


筛选小分子抑制剂：经验证，EGFR小分子抑制剂可以抑制EGF蛋白 (Cat. No. EGF-H52H3)诱导的报告基因活性，EC50值为0.01 μM。

Inhibition of human EGF protein-induced reporter activity by human EGFR small molecule inhibitor. This reporter cell was incubated with serial dilutions of inhibitors in the presence of human EGF protein (ACROBiosystems, Cat.No.EGF-H52H3) with a final concentration of 50 ng/mL. The EC50 of human EGFR small molecule inhibitor (Erlotinib) was approximately 0.01 μM.

点击申请Protocol



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