BLI (Biolayer interferometry) binding affinity measurements of Anti-Zolbetuximab Antibody (9C9), mAb, Mouse (GenScript, A02261) to Zolbetuximab.
Zolbetuximab captured on HFC (Anti-Human IgG Fc) Probes can bind Anti-Zolbetuximab Antibody (9C9), mAb, Mouse with a dissociation constant (KD) of less than 1 pM.

Anti-Zolbetuximab Antibody (34E11) or (9C9) inhibits the binding of Zolbetuximab to Claudin18.2 (CLDN18.2). Coating antigen: Zolbetuximab 1 µg/ml.
Claudin18.2 (GenScript, Z03504) final concentration: 1.2 µg/ml.
Anti-Zolbetuximab antibody dilutions start from 10 μg/ml.
Anti-Zolbetuximab Antibody (34E11), mAb, Mouse (GenScript, A02260) exhibited non-inhibitory properties, whereas Anti-Zolbetuximab Antibody (9C9), mAb, Mouse (GenScript, A02261) demonstrated significant inhibitory properties with an IC50 value of 0.44 µg/ml.

Standard curve of Zolbetuximab sandwich ELISA by A02260 (34E11) and A02261 (9C9).
In this ELISA assay, Anti-Zolbetuximab Antibody (34E11), mAb, Mouse (GenScript, A02260) was coated at a concentration of 1 μg/ml, and Anti-Zolbetuximab Antibody (9C9), mAb, Mouse (GenScript, A02261) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/ml.
In this ELISA assay, a four-parameter logistic curve fitting program was used to create a standard curve with the R-Square equal to 0.99984. The typicaldynamic range of the assay is 2-128 ng/mL and its sensitivity of detecting Zolbetuximab is up to 2 ng/ml.

Specificity analysis of the detection of Zolbetuximab.

Zolbetuximab standard samples at 8 concentrations were spiked with 2 µg/mL of human IgG1. The test result demonstrated that the high concentration of human IgG1 did not interfere with the detection of Zolbetuximab.
In this ELISA assay, Anti-Zolbetuximab Antibody (34E11), mAb, Mouse (GenScript, A02260) was coated at a concentration of 1 μg/ml, and Anti-Zolbetuximab Antibody (9C9), mAb, Mouse (GenScript, A02261) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/ml.

MRD analysis of the detection of Zolbetuximab in biological matrix.

The MRD is the minimum dilution necessary for the detection of Zolbetuximab in biological matrix with least interference. Serum samples from cynomolgus monkey were serially diluted to determine the MRD of this assay, and the test result suggestedthat MRD was as 1:20.
In this ELISA assay, Anti-Zolbetuximab Antibody (34E11), mAb, Mouse (GenScript, A02260) was coated at a concentration of 1 μg/ml, and Anti-Zolbetuximab Antibody (9C9), mAb, Mouse (GenScript, A02261) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/ml.