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Your Position: > Star Staining Fluorescent-labeled Products New Generation Tools for CAR Detection

Star Staining Fluorescent-labeled Products
New Generation Tools for CAR Detection

Star Staining-Fluorescent-labeled Products
Introduction
Star Staining fluorescent-labeled products are developed by a new-generation site-specific labeling technology with "Star Standard" quality at ACROBiosystems. These products are uniquely designed for detecting and monitoring CAR-T cells in clinical trials.
Fluorescent-labeled protein is a powerful tool for the detection of CAR expression in research and clinical samples by flow cytometry. These proteins are pre-labeled fluorescent dyes that can detect CAR expression by one-step staining with minimal background. However, the development of high-quality fluorescent-labeled proteins presents a major challenge and there are only few products commercially available so far.
Conjugation technique is a major bottleneck for developing high-quality fluorescent-labeled proteins. Currently, the most widely used labeling technique in the market is the traditional chemical labeling approach. It is a very easy method to covalently label protein with a fluorescent dye in a non-specific manner. However, random modification can lead to heterogeneous and may affect the bioactivity of the protein via blocking the protein-active site. Studies indicate that the new-generation site-specific labeling technology can attach fluorescent dyes to specific protein in site-specific manner. It provides a guarantee to the uniformity of the fluorescent-labeled products and avoids affecting protein activity. This property makes it a useful weapon for the development of high-quality fluorescent-labeled protein products with high specificity and sensitivity.
Product Feature
Traditional Chemical Labeling
Traditional Chemical Labeling
Non-selective random labeling and affect natural structure
May cover the active sites to reduce bioactivity
Easy to aggregate due to change the hydrophilicity of proteins
High variation in batch-to-batch consistency
New-generation Site-specific Labeling
New-generation Site-specific Labeling
The labeling happens at the specific tag and far away from active sites of proteins
Maintain natural conformation and modification
Efficiently bio-orthogonal technology
High batch-to-batch consistency and uniformity.
Star Staining Features

ACROBiosystems is striving for developing the high-standard detection reagents to support the development of CAR-T cell therapy. After years of research and experience by our technicians, we successfully established the "Star Staining" site-specific labeling platform and developed a series of exclusive fluorescent-labeled proteins with "Star Standard" quality. "Star Staining" products can be the powerful tools for detecting and monitoring CAR-T cells in clinical trials.

Using new-generation site-specific labeling technology to maintain natural bioactivity

High specificity and sensitivity verified by flow cytometry

No non-specific binding to non-transduced PBMCs

High batch-to-batch consistency and uniformity

Hot Products
  • FITC

  • PE

  • APC

  • Alexa Fluor 647

  • Alexa Fluor 555

  • Alexa Fluor 488

MoleculeCat. No.Product DescriptionPreorder/Order
MoleculeCat. No.Product DescriptionPreorder/Order
MoleculeCat. No.Product DescriptionPreorder/Order
MoleculeCat. No.Product DescriptionPreorder/Order
MoleculeCat. No.Product DescriptionPreorder/Order
MoleculeCat. No.Product DescriptionPreorder/Order
Data Display

High purity

High purity than 90% of Star Staining FITC-labeled Human BCMA
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.

High bioactivity

Higher binding activity than that of other competitors
BCMA binding activity verified by FACS
CD19 binding activity verified by FACS

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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Alexa Fluor 647-Labeled Human Mesothelin (296-580)

The activity of Alexa Fluor 647-Labeled Human Mesothelin (296-580), His Tag (Cat. No. MSN-HA2H5) was higher than other Competitor.


Alexa Fluor 488-Labeled Human Mesothelin / MSLN (296-580) Protein

The activity of Alexa Fluor 488-Labeled Human Mesothelin (296-580), His Tag (Cat. No. MSN-HA2H9) was higher than other Competitor.

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Star Staining PE-CD19 or APC-CD19 protein can detect CD19 CAR expression with high sensitivity by flow cytometry.
APC-CD19 FACS binding activity verified by FACS

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.

PE-CD19 binding activity verified by FACS

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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No non-specific binding to non-transduced PBMCs

Star Staining products have no non-specific binding to non-transduced PBMCs with higher binding activity than that of other competitors
BCMA-PBMC  verified by FACS

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 comparison Data

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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Maintain natural bioactivity

High binding capacity before and after conjugation, verified by FACS and SPR
BCMA bioactivity verified by FACS

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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BCMA binding affinity  verified by SPR
FITC-BCMA binding affinity  verified by SPR

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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High batch-to-batch consistency

Binding activity of different lots of FITC/Alexa Fluor-555/PE-labeled Human BCMA protein was verified by FACS
FITC-BCMA batch-to-batch consistency verified by FACS
AF555-BCMA  batch-to-batch consistency verified by FACS
PE-BCMA batch-to-batch consistency verified by FACS

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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High stability

PE-BCMA stability verified by FACS
FITC-BCMA stability verified by FACS

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

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More CAR-T Related Products

REFERENCES

[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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