CAR-T 타겟 단백질
Chimeric antigen receptor T (CAR-T) cell therapy is a cancer immunotherapy based on genetically modified T cells that express specific receptor fragments recognizing tumor surface antigens. The modified T cells are infused into patients and can directly target cancer cells in the body without the help of antigen-presenting cells (APCs) and exert immune killing effects.
For CAR-T cells, the effective component for tumor killing is CAR-positive T cells. The packaging specifications and clinical dosage of CAR-T cell products are expressed by the number of CAR-positive cells. Therefore, CAR transduction positive rate is a mandatory test item for CAR-T quality control. Regulatory authorities recommend using flow cytometry to detect CAR transduction positive rate. Currently, there are detection methods for different structural regions of CAR, including those targeting CAR antigen binding sites, such as CD19 antigen, or anti-Fab antibodies or Protein L proteins targeting light or heavy chain regions. Among them, the CAR positive rate detection method targeting antigen binding sites has been widely used due to its better specificity.
ACROBiosystems, as a protein supplier focused on pharmaceutical R&D, has developed a series of 형광 표기 단백질 in various forms including unlabeled, biotin-labeled, and fluorescent-labeled using professional protein development platform, protein labeling platform, stable cell line development platform and flow cytometry analysis platform, as well as supporting flow cytometry protocols for detecting CAR positive rates, to assist CAR-T R&D and accelerate the progress of CAR-T research. Currently, products cover more than 40 popular CAR-T targets including CD19, BCMA, CD22, MSLN and EGFR.
직접 검출법 - 형광 표기 단백질
검사 절차를 간소화하고 검사 시간을 절약할 수 있습니다
이항 교차 반응으로 인한 비특이성 배경을 피할 수 있습니다
- FITC를 사용하여 CD19 단백질을 표시하여 Anti-CD19-CAR의 양성률 측정
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- 293 cells were transfected with anti-CD19-scFv and RFP tag. 2e5 of the cells were stained with B. FITC-Labeled Human CD19 (20-291) (Cat. No. CD9-HF2H2, 10 µg/ml) and C. FITC-labeled protein control. A. Non-transfected 293 cells and C. 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) (Cat. No. CD9-HF2H2).
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Protocol
생물소-친화소를 바탕으로 하는 검측법-생물소 표기단백질
스트렙트아비딘은 생물소에 대해 매우 높은 친화력을 가지고 있으며, 이들의 결합은 고도의 특이성을 가지고 있습니다 .
높은 민감도와 특이성을 가지고 CAR 양성 표현률을 검출할 수 있습니다.
- 생물소 표기 BCMA 단백질을 사용하여 Anti-BCMA-CAR의 양성률 측정
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- Human T cells were transfected with anti-BCMA CAR and cultured for 3 days. Three days post-transfection, 1e6 cells were first incubated with 50 µl biotinylated human BCMA protein (Cat. No. BC7-H82F0, 8 µg/ml), washed and then stained with PE Streptavidin and analyzed by flow cytometry. (Data are kindly provided by PREGENE Biopharma)
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Protocol
간접 검출법 - 비표기 단백질
CAR 양성 발현률을 매우 민감하게 감지할 수 있습니다.
이항 교차 반응으로 인한 비특이성 배경이 생길 수 있습니다.
- Human CD19, Fc Tag 단백질을 사용하여 Anti-CD19-CAR 양성률 측정
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- 293 cells were transfected with FMC63-scFv and RFP tag. 2e5 of the cells were first stained with B. Human CD19 (20-291) Protein, Fc Tag, low endotoxin (Super affinity) (Cat. No. CD9-H5251, 3 µg/ml) and C. Human Fc Tag Protein Control, followed by FITC-conjugated Anti-human IgG Fc Antibody. A. Non-transfected 293 cells and C. Human Fc Tag 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 Human CD19 (20-291) Protein, Fc Tag, low endotoxin (Super affinity) (Cat. No. CD9-H5251).
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Request Protocol
- 형광 표기 단백질
- 직접 검출법 - 형광 표기 단백질
- 생물소-친화소를 바탕으로 하는 검측법-생물소 표기단백질
- 간접 검출법 - 비표기 단백질
- 더 많은 CAR-T 관련 내용