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Star Staining——新一代荧光定点标记技术平台及系列产品

Star Staining-新一代荧光定点标记技术平台及系列产品
背景
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”的技术平台及产品,
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产生的非特异背景,成为流式检测CAR表达的首选试剂。
然而,荧光标记蛋白的选择面临着诸多难题。一方面,由于市面上主流的蛋白标记技术为传统随机化学标记方法,易改变蛋白的天然结构,导致活性下降,降低检测灵敏度而无法检测低丰度的CAR,另外,使用随机化学标记技术,易造成蛋白聚集,在进行FACS检测时会产生非特异信号,且标记工艺不稳定导致荧光蛋白产品不均一,难以控制批间差,因此,使用传统随机化学标记方法,很难获得高活性、高批间一致性的荧光标记蛋白,从而影响CAR-T药物的质量控制,更难以用于临床分析应用;另一方面,在进行多重染色时,需要选择多种荧光染料的标记蛋白,或流式检测设备所限,仅能选择用于该设备所匹配的荧光,因此,丰富的荧光类型,也是FACS检测CAR表达时所考虑的要点。
为解决上述问题,ACROBiosystems凭借Star Staining新一代定点标记技术平台,成功开发了一系列荧光标记类型的CAR靶点相关蛋白产品,以“星级”的产品优势,全方位满足CAR-T阳性率检测的不同需求。Star Staining新一代定点标记技术平台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。
技术原理
传统化学标记技术
传统化学标标记技术
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
新一代定点标记技术
新一代定点标记技术
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
Star Staining特色

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。

采用新一代定点标记技术, 保持蛋白天然构象,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

热门产品
荧光类型 分子 货号 产品描述 订购/预购
FITCBCMABCA-HF2H3 FITC-Labeled Human BCMA / TNFRSF17 Protein, His Tag star staining

订购

CD19CD9-HF2H3 FITC-Labeled Human CD19 (20-291) Protein, His Tag star staining

订购

MesothelinMSN-HF2H3 FITC-Labeled Human Mesothelin / MSLN (296-580) Protein, His Tag star staining

订购

PEBCMABCA-HP2H7 PE-Labeled Human BCMA / TNFRSF17 Protein, His Tag star staining

订购

CD19CD9-HP2H5 PE-Labeled Human CD19 (20-291) Protein, His Tag star staining

订购

MesothelinMSN-HP2H8 PE-Labeled Human Mesothelin / MSLN (296-580) Protein, His Tag star staining

订购

Siglec-2 SI2-HP2H5 PE-Labeled Human Siglec-2 / CD22 Protein, His Tagstar staining

订购

APCBCMABCA-HA2H4 APC-Labeled Human BCMA / TNFRSF17 Protein, His Tag star staining

订购

CD19CD9-HA2H9 APC-Labeled Human CD19 (20-291) Protein, His Tag star staining

预购

MesothelinMSN-HA2H6 APC-Labeled Human Mesothelin / MSLN (296-580) Protein, His Tag star staining

预购

更多Star Staining 产品列表
  • FITC
  • PE
  • APC
  • Alexa Fluor 647
  • Alexa Fluor 555
  • Alexa Fluor 488
荧光类型 分子 产品描述 订购/预购
FITC BCMA FITC-Labeled Human BCMA / TNFRSF17 Protein, His Tagstar staining

订购

CD19 FITC-Labeled Human CD19 (20-291) Protein, His Tagstar staining

订购

Mesothelin FITC-Labeled Human Mesothelin / MSLN (296-580) Protein, His Tagstar staining

订购

Glypican 3 FITC-Labeled Human Glypican 3 / GPC3 Protein, His Tag star staining

预购

HER2 FITC-Labeled Human Her2 / ErbB2 Protein, His Tag star staining

预购

CD7 FITC-Labeled Human CD7 Protein, His Tag star staining

预购

Siglec-3 FITC-Labeled Human Siglec-3 / CD33 Protein, His Tag star staining

预购

CD30 FITC-Labeled Human CD30 / TNFRSF8 Protein, His Tag star staining

预购

EGF R FITC-Labeled Human EGF R Protein, His Tag star staining

预购

EGFRVIII FITC-Labeled Human EGFRvIII Protein, His Tag star staining

预购

GUCY2C FITC-Labeled Human GUCY2C / Guanylyl cyclase C Protein, His Tag star staining

预购

NKG2D FITC-Labeled Human NKG2D / CD314 Protein, His Tag star staining

预购

FAP FITC-Labeled Human FAP Protein, His Tag star staining

预购

Nectin-4 FITC-Labeled Human Nectin-4 Protein, His Tag star staining

预购

CD37 FITC-Labeled Human CD37 Protein, His Tag star staining

预购

protein L FITC-Labeled Recombinant Protein L Protein, His Tag star staining

预购

荧光类型 分子 产品描述 订购/预购
PE BCMA PE-Labeled Human BCMA / TNFRSF17 Protein, His Tag star staining

订购

CD19 PE-Labeled Human CD19 (20-291) Protein, His Tag star staining

订购

Mesothelin PE-Labeled Human Mesothelin / MSLN (296-580) Protein, His Tag star staining

订购

Glypican 3 PE-Labeled Human Glypican 3 / GPC3 Protein, His Tag star staining

预购

NKG2D PE-Labeled Human NKG2D / CD314 Protein, His Tag star staining

预购

FAP PE-Labeled Human FAP Protein, His Tag star staining

预购

Nectin-4 PE-Labeled Human Nectin-4 Protein, His Tag star staining

预购

CD37 PE-Labeled Human CD37 Protein, His Tag star staining

预购

protein L PE-Labeled Recombinant Protein L Protein, His Tag star staining

预购

EMMPRIN PE-Labeled Human EMMPRIN / CD147 Protein, His Tag star staining

预购

CD300e PE-Labeled Human CD300e Protein, His Tag star staining

预购

HER2 PE-Labeled Human Her2 / ErbB2 Protein, His Tag star staining

预购

Siglec-2 PE-Labeled Human Siglec-2 / CD22 Protein, His Tag star staining

订购

Siglec-3 PE-Labeled Human Siglec-3 / CD33 Protein, His Tag star staining

预购

PD-1 PE-Labeled Human PD-1 Protein, His Tag star staining

预购

OX40 PE-Labeled Human OX40 Protein, His Tag star staining

预购

Mucin-1 PE-Labeled Human MUC-1 Protein, His Tag star staining

预购

TSLP R PE-Labeled Human TSLP R Protein, His Tag star staining

预购

荧光类型 分子 产品描述 订购/预购
APC BCMA APC-Labeled Human BCMA / TNFRSF17 Protein, His Tag star staining

订购

CD19 APC-Labeled Human CD19 (20-291) Protein, His Tag star staining

预购

Mesothelin APC-Labeled Human Mesothelin / MSLN (296-580) Protein, His Tag star staining

预购

Glypican 3 APC-Labeled Human Glypican 3 / GPC3 Protein, His Tag star staining

预购

EMMPRIN APC-Labeled Human EMMPRIN / CD147 Protein, His Tag star staining

预购

CD300e APC-Labeled Human CD300e Protein, His Tag star staining

预购

HER2 APC-Labeled Human Her2 / ErbB2 Protein, His Tag star staining

预购

Siglec-2 APC-Labeled Human Siglec-2 / CD22 Protein, His Tag star staining

预购

Siglec-3 APC-Labeled Human Siglec-3 / CD33 Protein, His Tag star staining

预购

PD-1 APC-Labeled Human PD-1 Protein, His Tag star staining

预购

OX40 APC-Labeled Human OX40 Protein, His Tag star staining

预购

Mucin-1 APC-Labeled Human Mucin-1/MUC-1 Protein, His Tag star staining

预购

TSLP R APC-Labeled Human TSLP R Protein, His Tag star staining

预购

荧光类型 分子 产品描述 订购/预购
Alexa Fluor 647 BCMA Alexa Fluor 647-Labeled Human BCMA / TNFRSF17 Protein, His Tag star staining

预购

CD19 Alexa Fluor 647-Labeled Human CD19 (20-291) Protein, His Tag star staining

预购

Mesothelin Alexa Fluor 647-Labeled Human Mesothelin / MSLN (296-580) Protein, His Tag star staining

预购

Glypican 3 Alexa Fluor 647-Labeled Human Glypican 3 / GPC3 Protein, His Tag star staining

预购

EMMPRIN Alexa Fluor 647-Labeled Human EMMPRIN / CD147 Protein, His Tag star staining

预购

CD300e Alexa Fluor 647-Labeled Human CD300e Protein, His Tag star staining

预购

HER2 Alexa Fluor 647-Labeled Human Her2 / ErbB2 Protein, His Tag star staining

预购

Siglec-2 Alexa Fluor 647-Labeled Human Siglec-2 / CD22 Protein, His Tag star staining

预购

Siglec-3 Alexa Fluor 647-Labeled Human Siglec-3 / CD33 Protein, His Tag star staining

预购

PD-1 Alexa Fluor 647-Labeled Human PD-1 Protein, His Tag star staining

预购

OX40 Alexa Fluor 647-Labeled Human OX40 Protein, His Tag star staining

预购

Mucin-1 Alexa Fluor 647-Labeled Human Mucin-1/MUC-1 Protein, His Tag star staining

预购

TSLP R Alexa Fluor 647-Labeled Human TSLP R Protein, His Tag star staining

预购

荧光类型 分子 产品描述 订购/预购
Alexa Fluor 555 BCMA Alexa Fluor 555-Labeled Human BCMA / TNFRSF17 Protein, His Tag star staining

预购

CD19 Alexa Fluor 555-Labeled Human CD19 (20-291) Protein, His Tag star staining

预购

Mesothelin Alexa Fluor 555-Labeled Human Mesothelin / MSLN (296-580) Protein, His Tag star staining

预购

Glypican 3 Alexa Fluor 555-Labeled Human Glypican 3 / GPC3 Protein, His Tag star staining

预购

NKG2D Alexa Fluor 555-Labeled Human NKG2D / CD314 Protein, His Tag star staining

预购

FAP Alexa Fluor 555-Labeled Human FAP Protein, His Tag star staining

预购

Nectin-4 Alexa Fluor 555-Labeled Human Nectin-4 Protein, His Tag star staining

预购

CD37 Alexa Fluor 555-Labeled Human CD37 Protein, His Tag star staining

预购

protein L Alexa Fluor 555-Labeled Recombinant Protein L Protein, His Tag star staining

预购

荧光类型 分子 产品描述 订购/预购
Alexa Fluor 488 BCMA Alexa Fluor 488-Labeled Human BCMA / TNFRSF17 Protein, His Tag star staining

预购

CD19 Alexa Fluor 488-Labeled Human CD19 (20-291) Protein, His Tag star staining

预购

Mesothelin Alexa Fluor 488-Labeled Human Mesothelin / MSLN (296-580) Protein, His Tag star staining

预购

Glypican 3 Alexa Fluor 488-Labeled Human Glypican 3 / GPC3 Protein, His Tag star staining

预购

EMMPRIN Alexa Fluor 488-Labeled Human EMMPRIN / CD147 Protein, His Tag star staining

预购

CD300e Alexa Fluor 488-Labeled Human CD300e Protein, His Tag star staining

预购

HER2 Alexa Fluor 488-Labeled Human Her2 / ErbB2 Protein, His Tag star staining

预购

Siglec-2 Alexa Fluor 488-Labeled Human Siglec-2 / CD22 Protein, His Tag star staining

预购

Siglec-3 Alexa Fluor 488-Labeled Human Siglec-3 / CD33 Protein, His Tag star staining

预购

PD-1 Alexa Fluor 488-Labeled Human PD-1 Protein, His Tag star staining

预购

OX40 Alexa Fluor 488-Labeled Human OX40 Protein, His Tag star staining

预购

Mucin-1 Alexa Fluor 488-Labeled Human Mucin-1/MUC-1 Protein, His Tag star staining

预购

TSLP R Alexa Fluor 488-Labeled Human TSLP R Protein, His Tag star staining

预购

验证数据

纯度高于90%

Star Staining FITC-labeled Human BCMA产品纯度高于90%。
BCMA SDS-PAGE

FITC-Labeled Human BCMA, His Tag (Cat. No. BCA-HF2H3) on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.

BCMA SEC-MALS

The purity of FITC-Labeled Human BCMA, His Tag (Cat. No. BCA-HF2H3) is more than 90% and the molecular weight of this protein is around 24-34 kDa verified by SEC-MALS.

高灵敏度

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

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 (Cat. No. BCA-HF2H3) and CD19 (Cat. No. CD9-HF2H3) protein have a much higher binding activity than that of the other competitor.

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Star Staining PE-labeled 和APC-labeled CD19蛋白产品经流式验证可以高灵敏度检测CD19 CAR表达。
PE-CD19  灵敏度FACS验证数据图

Anti-CD19 CAR-293 cells were stained with PE-Labeled Human CD19 (20-291) Protein (Cat. No. CD9-HP2H5), His Tag star staining and negative control protein respectively, PE signal was used to evaluate the binding activity.

APC-CD19  灵敏度FACS验证数据图

Anti-CD19 CAR-293 cells were stained with APC-Labeled Human CD19 (20-291) Protein (Cat. No. CD9-HA2H9), His Tag star staining and negative control protein respectively, APC signal was used to evaluate the binding activity.

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极低非特异背景

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。
BCMA-PBMC  FACS验证数据图

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

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BCMA PBMC 验证对比图

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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可保持蛋白天然活性

Star Staining FITC-labeled Human BCMA产品可保持天然蛋白活性。
FACS验证结果显示标记FITC前后BCMA蛋白天然活性保持不变
BCMA活性验证FACS数据图

Binding activity of the Human BCMA before and after FITC labeling was evaluated in the above FACS analysis. The result shows that FITC-Labeled BCMA (Cat. No. BCA-HF2H3) and unconjugated Human BCMA have almost the same level of binding activity.

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SPR验证结果显示标记FITC前后BCMA蛋白的亲和力水平无差异
BCMASPR亲和力验证数据图
FITC-BCMASPR亲和力验证数据图

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 (Cat. No. BCA-HF2H3) and unconjugated Human BCMA, His Tag have almost the same level of affinity.

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高批间一致性

不同荧光标记产品的不同批次蛋白活性可保持一致,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。
FITC-BCMA批间一致性FACS验证数据图
AF555-BCMA批间一致性FACS验证数据图
PE-BCMA批间一致性FACS验证数据图

Binding activity of different lots of FITC-Labeled Human BCMA (Cat. No. BCA-HF2H3) , AF555-Labeled Human BCMA (Cat. No. BCA-HA2H6) and PE-Labeled Human BCMA (Cat. No. BCA-HP2H7) 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

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。
PE-BCMA高稳定性FACS验证数据图
FITC-BCMA高稳定性FACS验证数据图

PE-Labeled Human BCMA (Cat. No. BCA-HP2H7) and FITC-Labeled Human BCMA (Cat. No. BCA-HF2H3) at 25℃ for 48 hours and freeze-throw cycles are stable without performance reduction.

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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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