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Application: Study complex formation and screen simultaneously the BsAbs binding affinity towards dual-antigen targets for drug discovery and high throughput screening (HTS) applications. Note: Suitable for use with human serum, cell culture supernatants, and with purified proteins.
Background: Bispecific antibodies (BsAbs) are antibodies that have binding sites directed against different antigens or different epitopes of the same antigen. They are composed of heavy and light chains, and the appropriate chain matching is required for efficacy. To enhance the chance of chain matching and different applications, more than 30 different technologies have been developed, such as knob-into hole, ART-Ig, BiTE (bispecific cell engager) and others. They have gained attention for the treatment of cancer and other disorders due to their superior cytotoxicity effects and less development of resistance to their use. BsAbs can act as immune cell-cancer cell bridges, enhancing the killing potential of the immune cells. Current BsAbs have a CD3, CD16 or CD47 as one of the binding domains, and target CD19, PD-1, LAG-3, PSMA, mesothelin or other tumor antigens with the other. For instance, B-lymphocyte antigen CD19 (Cluster of Differentiation 19), also known as B-Lymphocyte Surface Antigen B4 and CVID3C, is a transmembrane protein expressed in follicular dendritic cells and all B lineage cells, and is involved in the balance of humoral, antigen-induced and tolerance responses. Its presence in normal and neoplasmic B cells has made it an attractive target in cancer therapy. Blinatumomab is a CD19/CD3 BiTE already approved by the FDA for use in patients with refractory or relapsed B-cell precursor ALL (acute lymphoblastic leukemia), and now being considered in CLL (chronic lymphocytic leukemia) treatment. The development of BsAbs targeting new targets will advance multiple biomedical research field.
Contraindications: This kit is compatible with up to 1% DMSO.
Description: The Protein Of Interest (POI): CD3 Bridging Chemiluminescent Assay Kit is an ELISA designed to analyze the ability of a Bispecific Antibody (BsAbs) to POI and CD3 (cluster of differentiation 3) for screening and profiling applications. This assay allows to study the simultaneous binding to CD3 and to a protein/antigen of interest of bispecific antibodies. The protein of interest must be provided by the user. The Protein of Interest (POI): CD3 Bridging Chemiluminescent Assay Kit comes with enough CD3-Containing Detection Reagent, assay buffers and controls (reference capture antigen (BCMA protein) and reference bispecific antibody (targeting BCMA)) for 100 reactions. This kit provides the user with the flexibility to test their own specific POI, compare variants, or assay several POIs at once for specific determination or epitope mapping, using an optimized protocol and CD3 reagent. The native state of the POI, optimal concentration to be used in plate coating, and ability to be recognized by the test antibodies should be assessed prior the testing with this assay kit.Figure 1: Protein Of Interest (POI): CD3 Bridging Chemiluminescent Assay Kit assay principleFirst a 96-well plate is coated with POI and blocked. Next, bispecific antibodies are added in an optimized assay buffer. This is followed by washing to remove any unbound bispecific antibodies, and the plate is incubated with CD3-Containing Detection Reagent. Finally, ELISA ECL substrate is added and the chemiluminescence generated can be measured using a chemiluminescence reader. The chemiluminescence signal is proportional to the efficacy of the bridging.Need us to run inhibitor screens or profile your bispecific antibodies towards CD3 and BCMA? Check out our Immunotherapy Biochemical Screening Services.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Target: CD3
Uniprot: Q02223
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References: Robinson H., et al., 2018 Blood 132 (5): 521-532.Ma J., et al., 2021 Front Immunol. 12:626616.
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