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Ideal Tools for Anti-CD19 CAR Expression

ACRO

CD19 protein

Detect Anti-CD19 CAR Expression by FACS - 1e6 of the anti-CD19 CAR-293 cells were stained with 100 μL of 1:50 dilution (2 μL stock solution in 100 μL FACS buffer) of PE-Labeled Human CD19 (20-291), His Tag (SKU No. CD9-HP2H3). PE Streptavidin was used as negative control (QC tested).

The affinity constant of human CD19 protein was 0.255 nM as verified by SPR - Biotinylated Human CD19 (20-291), His,Avitag (Cat. No. CD9-H82E9) captured on Biotin CAP - Series S sensor Chip can bind FMC63 MAb (mouse lgG2a) with an affinity constant of 0.255 nM as determined in a SPR assay (Biacore 8K) (QC tested)

Higher Binding Affinity than other vendors as verified by FACS and ELISA - Binding activity of FITC-Labeled Human CD19, His Tag from two different vendors were evaluated in the flow cytometry analysis against anti-CD19-CAR-293 cells above. The result showed that ACRO's FITC-Labeled Human CD19, His Tag has a much higher binding activity than that of the other vendor.

Higher Binding Affinity than other vendors as verified by FACS and ELISA - Binding activity of FITC-Labeled Human CD19, His Tag from two different vendors were evaluated in the ELISA analysis against FMC63 MAb above. The result showed that ACRO's FITC-Labeled Human CD19, His Tag has a much higher binding activity than that of the other vendor.

Anti-FMC63 scFv Antibody

Detect Anti-CD19 (FMC63) CAR Expression by FACS - 2e5 of Anti-CD19 CAR-293 cells were stained with 100 μL of 1:50 dilution (2 μL stock solution in 100 μL FACS buffer) FITC-Labeled Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 (Cat. No. FM3-FY45) and isotype control respectively. FITC signal was used to evaluate the binding activity (QC tested).

Binding Activity and Specificity Measured by ELISA with a linear range of 1-19 ng/mL - Immobilized FMC63 scFv, His Tag at 2 μg/mL (100 μL/well) can bind Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 (Cat. No. FM3-Y45) with a linear range of 1-19 ng/mL. Anti-DNP antibody, mouse IgG1 (Cat. No. DNP-M1) was used as an isotype control (QC tested).

Neutralizing Activity Measured by ELISA, IC50= 0.084 μg/mL - ELISA analysis shows that the binding of Human CD19, Fc Tag (Cat. No. CD9-H5251) to FMC63 scFv, His Tag was inhibited by increasing concentration of Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 (Cat. No. FM3-Y45). The concentration of Human CD19, Fc Tag used is 5 μg/mL (100 μL/well). The IC50 is 0.084 μg/mL (Routinely tested).

High Affinity Determined by SPR - Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 (Cat. No. FM3-Y45) captured on CM5 chip via anti-mouse antibodies surface can bind FMC63 scFv with an affinity constant of 1.08 nM as determined in a SPR assay.

HEK293 Expressed Full Length CD20 Protein

CD20 is a hot target for the treatment of lymphoma, leukemia and some autoimmune diseases. However, due to low expression titer and the unstable bioactivity, it is so difficult to find an active CD20 protein on the market for antibody drug development and the CD20 CAR expression detection.

ACROBiosystems has developed the highly active full length multi-transmembrane CD20 proteins, expressed by HEK293 cells. The active full length CD20 proteins carry both big and small ECD loops, and bind Rituximab and Ofatumumab with high binding affinity in ELISA and SPR assay. We have different types of CD20 proteins include unconjugated protein, biotinylated proteins with DDM and Nanodisc, which are suitable for immunization, ELISA, SPR, BLI and FACS.

Product Features

  • Authentic conformation with both small and big loops
  • Super bioactivity verified by ELISA/SPR/BLI/FACS
  • Suitable for drug screening, characterization, quality control and CAR expression detection

 

Product: Biotinylated Human CD20 / MS4A1Full Length Protein, His,Avitag™ (Nanodisc) (HEK293).

Bioactivity-FACS 

2e5 of CD20-CAR-293 cells transfected with anti-CD20-scFv were stained with 100 μL of 3 μg/mL of Biotinylated Human CD20 Full Length, His,Avitag (SKU No. CD0-H82E3) and negative control antibody respectively, washed and then followed by PE-SA and analyzed with FACS.

Product: Human CD20 / MS4A1 Full Length Protein, His Tag (Nanodisc) (HEK293)

 

Bioactivity-FACS

 

2e5 of CD20-CAR-293 cells transfected with anti-CD20-scFv were stained with 100 μL of 3 μg/mL of Human CD20 Full Length, His Tag (SKU No. CD0-H52H1) and negative control antibody respectively, washed and then followed by PE anti-His antibody and analyzed with FACS.

Product: Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (HEK293)

 

Bioactivity-ELISA

 

Immobilized Rituximab at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (HEK293) (Cat. No. CD0-H82E5) with a linear range of 4-63 ng/mL (in presence of DDM and CHS).

Product: Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (HEK293)

 

Bioactivity-ELISA

 

Immobilized Ofatumumab at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (HEK293) (Cat. No. CD0-H82E5) with a linear range of 4-63 ng/mL (in presence of DDM and CHS).

Product: Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (HEK293)

 

Bioactivity-SPR

 

Biotinylated Human CD20, His,Avitag (HEK293) (Cat. No. CD0-H82E5) captured on Biotin CAP-Series S Sensor Chip can bind Rituximab with an affinity constant of 1.73 nM as determined in a SPR assay (in presence of DDM and CHS) (Biacore T200).

Product: Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (HEK293)

 

Bioactivity-SPR

 

Biotinylated Human CD20, His,Avitag (HEK293) (Cat. No. CD0-H82E5) captured on Biotin CAP-Series S Sensor Chip can bind Ofatumumab (Human IgG1) with an affinity constant of 1.03 nM as determined in a SPR assay (in presence of DDM and CHS) (Biacore T200)

 

Product: Human CD20 / MS4A1 Full Length Protein, His Tag (HEK293) (SPR verified)

 

Bioactivity-ELISA

 

Immobilized Rituximab at 2 μg/mL (100 μL/well) can bind Human CD20 Full Length, His Tag, HEK293 (SPR verified) (Cat. No. CD0-H52H3) with a linear range of 0.4-3 ng/mL (in presence of DDM and CHS)

 

Product: Human CD20 / MS4A1 Full Length Protein, His Tag (HEK293) (SPR verified)

 

Bioactivity-ELISA

 

Immobilized Ofatumumab at 2 μg/mL (100 μL/well) can bind Human CD20 Full Length, His Tag, HEK293 (SPR verified) (Cat. No. CD0-H52H3) with a linear range of 0.4-6 ng/m� (in presence of DDM and CHS).

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