Soluciones para la evaluación de la expresión del RAQ
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 CAR-T target proteins 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.
Método directo: proteínas marcadas con fluorescencia
Processing time can be reduced by the use of direct-labeled proteins.
Non-specific reaction of a secondary antibody is eliminated.
- Evaluation of Anti-CD19 CAR Expression with FITC-labeled CD19
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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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Detección basada en biotina-estreptavidina usando proteínas biotiniladas
Streptavidin labeled with fluorochromes can bind biotinylated proteins with a high degree of affinity and specificity, amplifying the signal and improving the detection sensitivity and specificity.
- Evaluation of Anti-BCMA CAR Expression with Biotinylated BCMA
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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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Detección indirecta usando proteínas no conjugadas
High sensitivity detection of CAR positive expression rate.
Non-specific reaction of a secondary antibody may occur.
- Evaluation of Anti-CD19 CAR Expression with Fc-fusion CD19
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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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Más productos relacionados con RAQ-T
- Background
- Método directo: proteínas marcadas con fluorescencia
- Detección basada en biotina-estreptavidina usando proteínas biotiniladas
- Detección indirecta usando proteínas no conjugadas
- Más productos relacionados con RAQ-T