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Application: Binding studies to mesothelin and CD3 in biochemical and cellular assays.
Assay Conditions: The antibody was validated by measuring anti-Mesothelin-anti-CD3 binding to Mesothelin antigen in ELISA. The anti-Mesothelin-anti-CD3 antibody (BPS Bioscience # 101621) was coated onto a 96-well plate overnight at 4°C (50 µl/well at a concentration of 4 µg/ml in PBS). The plate was washed 3 times with Immuno Buffer 1 (BPS Bioscience #79311) and blocked using 100 µl of Blocking Buffer 2 (BPS Bioscience #79728) for 1 hour at room temperature. After removing the blocking buffer, 50 µl/well of purified biotinylated Mesothelin protein (BPS Bioscience #100291), serially diluted in Blocking Buffer 2, was added for 30 minutes at room temperature. The plate was washed, incubated with Streptavidin-HRP, washed again, and incubated with the Colorimetric HRP substrate. The reaction was stopped, and absorbance was read at 450 nm. The Blank value was subtracted from all values.
Background: Bispecific antibodies (BsMab) are chimeric antibodies with the ability to bind to two antigens and are typically created for immune cell activation, interfering with signal pathways or bringing proteins together. Mesothelin is a tumor differentiation antigen present in mesothelioma, ovarian cancer, pancreatic adenocarcinoma and others. CD3 (Cluster of Differentiation 3) is present in T cells and is involved in T cell activation. The ability to bring cancer cells to T cells allows for enhanced tumor cell killing and is a valuable tool in cancer immunotherapy.
Description: Anti-mesothelin-Anti-CD3 (Knot-in-Knob) antibody recognizes human mesothelin and CD3 proteins. This human bispecific antibody has been tested for specific binding affinity to mesothelin. The recombinant antibody was affinity purified.
Format: Aqueous buffer solution.
Formulation: 8 mM phosphate, pH 7.4, 110 mM NaCl, 2.2 mM KCl, and 20% glycerol
Host Cell Line: HEK293
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Uniprot: Q13421
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)