Evaluation of CAR Expression - Protein Solutions

Fluorescence-Labeled Proteins

Key Features

• Specifically targets pre-labeled antigens with a fluorescent dye
• Significantly reduces processing time by using direct-labeled proteins
• Eliminates non-specific reactions of secondary antibodies

Image credit: ACRO Biosystems

FITC-Labeled

PE-Labeled

• MSLN

Case Study

Evaluation of Anti-CD19 CAR Expression with FITC-labeled CD19

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). Image credit: ACRO Biosystems

Biotinylated Proteins

Key Features

• Antigens pre-labeled with biotin can be specifically targeted and detected by labeled streptavidin, which is the biotin-avidin complex.
• Molecules labeled with streptavidin fluorochromes can bind biotinylated proteins with high affinity and specificity. This amplifies the signal and improves the overall sensitivity and specificity of the detection process.

Image credit: ACRO Biosystems

Case Study

Evaluation of Anti-BCMA CAR Expression with Biotinylated BCMA

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). Image credit: ACRO Biosystems

Unconjugated Proteins

Key Features

• Specifically targets antigens that have been designed to carry a specific tag
• Antigens are detected using a secondary antibody, which is also referred to as an anti-epitope tag antibody, that has been labeled with a fluorophore
• Non-specific reactions of a secondary antibody may occur

Image credit: ACRO Biosystems

Case Study

Evaluation of Anti-CD19 CAR Expression with Fc-fusion CD19

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). Image credit: ACRO Biosystems