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>技术资源 > 前沿速递 >【前沿进展】Star Staining!点亮CAR表达检测

【前沿进展】Star Staining!点亮CAR表达检测

2021-09-18 13:38    浏览量:1103

ACROBiosystems百普赛斯Star Staining新一代定点标记技术平台开发的高质量荧光标记蛋白,可保持蛋白的天然构象,不影响蛋白活性,均一性好、批间一致性高。“星级”的研发实力,“星级”的技术平台,产出“星级”的产品,Star Staining专为CAR-T细胞质控和临床样本分析检测而设计。


流式细胞术是细胞治疗药物研发过程中检测CAR阳性T细胞的重要技术手段,荧光标记蛋白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,成为流式检测CAR表达的首选试剂。

荧光标记技术是开发高质量荧光标记蛋白的关键。目前市面上主流的标记技术因采用传统随机化学标记方法,无法保证标记荧光的均一性且标记位点可能会覆盖蛋白的活性位点而影响蛋白本身活性,很难获得高批间一致性、高活性的荧光标记蛋白。研究显示,采用新一代荧光定点标记技术,可对蛋白的特定位点进行标记,标记速度快效率高,不影响蛋白的天然空间构象,活性好检测灵敏度高,产物均一性好,无非特异信号,这种独特的定点标记技术成为开发高质量荧光标记蛋白产品的有力武器。

传统化学标记 VS 定点标记


传统化学标记

  • 非选择性随机标记,影响蛋白正常结构

  • 蛋白活性位点被覆盖

  • 改变蛋白亲水性,蛋白易聚集

  • 产品批间差异大









新一代定点标记


  • 蛋白特定位点标记

  • 标记位点远离活性中心,维持蛋白天然空间构象

  • 高效生物正交反应,无非特异信号

  • 产物均一性好、批间一致性高


ACROBiosystems百普赛斯致力于开发适用于细胞治疗产品质控和临床样本分析的更高质量标准检测试剂产品,在多年技术经验积累的基础上,不断更新完善,以“星级”研发实力,成功搭建了“星级标准”的Star Staining新一代荧光定点标记技术平台,开发了独具特色的Star Staining系列荧光定点标记蛋白产品,助力细胞治疗药物的研发进程。



Star Staining特色



  • 采用新一代定点标记技术, 保持蛋白高活性

  • 高灵敏度和高特异性,经类CAR细胞批检验证

  • 极低非特异信号,经PBMC双染批检放行

  • 高度均一和批间一致性,高水平满足临床样本分析检测要求


Star Staining到底优秀到哪方面呢?看了下面的数据,您就明白啦~

>>>>Star Staining FITC-labeled Human BCMA产品纯度高于95%



>>>>Star Staining FITC-labeled Human BCMA和CD19蛋白产品的检测灵敏度显著高于竞品。


The binding activity of FITC-Labeled Human BCMA and CD19 protein from two different vendors were evaluated by the FACS analysis. The result showed that ACRO's Star Staining FITC-Labeled Human BCMA and CD19 protein have a much higher binding activity than that of the other competitor.

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>>>>Star Staining系列产品经未转染CAR的PBMC细胞双染批检测验证,无非特异背景问题,特异性显著高于竞品,保证CAR表达检测结果的特异性和准确性,适用于CAR-T临床样本分析。

Non-specific binding to non-transduced PBMCs between FITC-Labeled Human BCMA Protein of Acro and competitor. 5e5 of non-transduced PBMCs were stained with FITC-Labeled Human BCMA Protein and anti-CD3 antibody, washed and then analyzed with FACS. PE signal was used to evaluate the expression of CD3+ T cells in non-transduced PBMCs, and FITC signal was used to evaluate the non-specific binding activity to non-transduced PBMCs.

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>>>>Star Staining FITC-labeled Human BCMA产品可保持天然蛋白活性。

1. FACS验证结果显示标记FITC前后BCMA蛋白天然活性保持不变。


The binding activity of the Human BCMA before and after FITC labeling was evaluated in the above FACS analysis. The result shows that FITC-Labeled and unconjugated Human BCMA, His Tag have almost the same level of binding activity.

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2. SPR验证结果显示标记FITC前后BCMA蛋白的亲和力水平无差异。

The binding affinity of the Human BCMA before and after FITC labeling was evaluated in the above SPR analysis (Biacore T200). The result shows that FITC-Labeled and unconjugated Human BCMA, His Tag have almost the same level of affinity.

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>>>> Star Staining FITC-labeled Human BCMA产品不同批次、不同规格的蛋白活性保持一致,具有高批间一致性,以确保长期用于CAR-T细胞质控和临床样本分析中检测结果的稳定性和可靠性。

Binding activity of two different sizes (left) or three different lots(right) of FITC-Labeled Human BCMA against anti-BCMA CAR-293 cells was evaluated by flow cytometry. The result shows very high batch-to-batch consistency.

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>>>>高稳定性:25℃条件下加速48h和反复冻融三次,依然可保持较高的生物活性,显示出蛋白的高度稳定性,可放心储存和使用。

FITC-Labeled Human BCMA (Cat. No. BCA-HF2H3) in different concentrations at 25℃ for 48 hours and freeze-throw cycles are stable without performance reduction.

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Star Staining热门产品列表


点击图片查看产品详细信息

更多相关产品

>50+CAR-T靶点蛋白

>PE标记产品

>CD19系列产品

>FMC63系列产品


参考文献:

[1] Abbasov ME, Kavanagh ME, Ichu TA, et al. A proteome-wide atlas of lysine-reactive chemistry [published online ahead of print, 2021 Sep 9]. Nat Chem. 2021;10.1038/s41557-021-00765-4.

[2] Xu L, Kuan SL, Weil T. Contemporary Approaches for Site-Selective Dual Functionalization of Proteins. Angew Chem Int Ed Engl. 2021;60(25):13757-13777.

[3] Devabhaktuni A, Lin S, Zhang L, et al. TagGraph reveals vast protein modification landscapes from large tandem mass spectrometry datasets. Nat Biotechnol. 2019;37(4):469-479.


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