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Application: Study Claudin-18.2:CD3 complex formation and assess the simultaneous binding of BsAbs to their dual-antigen targets for drug discovery and high throughput screening (HTS).
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, Claudin-18.2 or other tumor antigens with the other. Claudin-18.2 is highly expressed in gastric and pancreatic adenocarcinoma and may be involved in tumor development and progression. Due to the morphological changes and epithelial-mesenchymal transition occurring upon cancer progression, the extracellular loops of claudin-18.2 become exposed and available for antibody binding. These biological characteristics suggest that it is an ideal target for therapy and have led to the further development of monoclonal antibodies against claudin-18.2, such as Zolbetuximab.
Contraindications: This kit is compatible with up to 1% DMSO.
Description: The Bispecific Claudin-18.2:CD3 Bridging Colorimetric ELISA Kit is an ELISA designed to analyze the ability of Bispecific Antibodies (BsAbs) to bridge Claudin-18.2 (Claudin-18, Isoform 2) and CD3 (cluster of differentiation 3) for screening and profiling applications. This assay kit can determine if an anti-Claudin-18.2-anti-CD3 bispecific antibody binds to both targets simultaneously and bridges Claudin-18.2 to CD3. The Bispecific Claudin-18.2:CD3 Bridging Colorimetric ELISA Assay Kit comes with enough recombinant human Claudin-18.2, CD3- Containing Detection Reagent, and assay buffer for 100 enzyme reactions. This kit also includes Anti-Claudin-18.2-Anti-CD3 Bispecific Antibody as a positive control.Figure 1: Bispecific Claudin-18.2:CD3 Bridging Colorimetric ELISA Assay Kit principle.A 96-well plate is coated with Claudin-18.2 protein. After coating, an anti-Claudin-18.2-anti-CD3 bispecific antibody is added in an optimized assay buffer. The plate is washed to remove the unbound antibody. The plate is then incubated with CD3-Containing Detection Reagent. Finally, HRP Colorimetric Substrate is added to produce absorbance that can be measured using a UV/Vis spectrophotometer microplate reader. The absorbance signal is proportional to the bridging efficacy.
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, Claudin-18
Uniprot: P56856
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References: Ma J., et al., 2021 Front Immunol. 12: 626616.
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