BLI (Biolayer interferometry) binding affinity measurements of MonoRab™ Anti-Certolizumab Antibody (12A3), mAb, Rabbit (GenScript, A02269) to Certolizumab.
MonoRab™ Anti-Certolizumab Antibody (12A3), mAb, Rabbit captured on Protein G Probes can bind Certolizumab pegol with a dissociation constant (KD) of 6.62E-10M.

Anti-Certolizumab Antibody (15H11) or (12A3) inhibits the binding of Certolizumab pegol to TNF-alpha.

Coating antigen: TNF-alpha (GenScript, Z01001), 1 µg/mL.
Certolizumab pegol final concentration: 0.25 µg/mL.
Anti-Certolizumab antibody dilutions start from 30 μg/mL.
MonoRab™ Anti-Certolizumab Antibody (15H11), mAb, Rabbit (GenScript, A02268) demonstrated weak inhibitory properties, whereas MonoRab™ Anti-Certolizumab Antibody (12A3), mAb, Rabbit (GenScript, A02269) demonstrated significant inhibitory properties, with an IC₅₀ value of 1.3 µg/mL.

Standard curve and specificity analysis of the detection of Certolizumab pegol by A02268 (15H11) and A02269 (12A3).

Certolizumab pegol standard samples at 8 concentrations were spiked with 5 µg/mL of human IgG1. The test result demonstrated that the high concentration of human IgG1 did not interfere with the detection of Certolizumab pegol.
In the ELISA assay shown in this chart, MonoRab™ Anti-Certolizumab Antibody (15H11), mAb, Rabbit (GenScript, A02268) was coated at a concentration of 2 μg/mL, and MonoRab™ Anti-Certolizumab Antibody (12A3), mAb, Rabbit (GenScript, A02269) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/mL.
In this assay, a four-parameter logistic curve fitting program was used to create a standard curve with the R-Square is greater than 0.999. The typicaldynamic range of the assay is 1.95-125 ng/mL and its sensitivity of detecting Certolizumab pegol is up to 1.95 ng/mL.

Standard curve and specificity analysis of the detection of Certolizumab pegol by A02269 (12A3) and anti-PEG antibody.

Certolizumab pegol standard samples at 8 concentrations were spiked with 5 µg/mL of human IgG1. The test result demonstrated that the high concentration of human IgG1 did not interfere with the detection of Certolizumab pegol.
In the ELISA assay shown in this chart, THE™ PEG Antibody, mAb, Mouse (GenScript, A01795) was coated at a concentration of 2 μg/ml, and MonoRab™ Anti-Certolizumab Antibody (12A3), mAb, Rabbit (GenScript, A02269) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/mL.
In this assay, a four-parameter logistic curve fitting program was used to create a standard curve with the R-Square is greater than 0.999. The typicaldynamic range of the assay is 0.55-400 ng/mL and its sensitivity of detecting Certolizumab pegol is up to 0.55 ng/mL.

MRD analysis of the detection of Certolizumab pegol in biological matrix by A02269 (12A3) and anti-PEG antibody.

The MRD is the minimum dilution necessary for the detection of Certolizumab pegol 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 1:10.
In the ELISA assay shown in this chart, THE™ PEG Antibody, mAb, Mouse (GenScript, A01795) was coated at a concentration of 2 μg/ml, and MonoRab™ Anti-Certolizumab Antibody (12A3), mAb, Rabbit (GenScript, A02269) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/mL.