CAR-specific Monoclonal Antibody to Detect Anti-CD20 CAR Expression

CD20 is an attractive target for CAR-T cell therapy in the treatment of B-cell malignancies, and it is also the target for rituximab. Rituximab is the first B-cell targeting therapeutic antibody approved by the US FDA in 1997. Recently, more and more scientists tried to construct their CD20 targeting CARs based on the scFv derived from rituximab. However, it is difficult to find an appropriate reagent for the evaluation of anti-CD20 (rituximab) CAR expression on the market.

To solve this issue, ACROBiosystems has developed the anti-rituximab monoclonal antibodies, which can recognize anti-CD20 (rituximab) CAR with high sensitivity and high specificity. The performance of anti-rituximab monoclonal antibodies were validated by flow cytometry (FCM) in house. And the antibodies are suit for detecting Anti-CD20 (rituximab) CAR expression. ACRO can provide the corresponding protocols for free.

An ideal reagent for the detection of Anti-CD20 (Rituximab) CAR expression

Evaluation of Anti-CD20 (Rituximab) Expression by FCM

2×10⁵ Anti-CD20 (Rituximab)-CAR 293 cells were first stained with anti-rituximab antibody (mouse IgG1, Cat. No. RIB-Y35) and followed by incubation with PE-labeled anti-mouse IgG1 antibody. PE-labeled anti-mouse IgG1 antibody was used as a negative control.

Protocol

Specifically recognize the antigen-binding site of Anti-CD20 (Rituximab) CAR with high affinity

Neutralizing activity measured by FCM

Flow Cytometry analysis shows that the binding of rituximab to 293F overexpressing CD20 was inhibited by increasing concentration of anti-rituximab antibodies (Cat. No. RIB-Y35). The concentration of rituximab used is 10 ng/ml. The IC50 is 0.013 μg/ml.

Protocol

High affinity determined by SPR

Anti-rituximab antibodies (mouse IgG1, Cat. No. RIB-Y35) captured on CM5 chip via anti-mouse antibodies surface, can bind rituximab with an affinity constant of 0.03 nM.

Protocol