BLI (Biolayer interferometry) binding affinity measurements of MonoRab™ Anti-Vedolizumab Antibody (76H8), mAb, Rabbit (GenScript, A02325) to Vedolizumab.
Vedolizumab captured on HFC (Anti-Human IgG Fc) Probes can bind MonoRab™ Anti-Vedolizumab Antibody (76H8), mAb, Rabbit with a dissociation constant (KD) of 1.05E-11M.

Anti-Vedolizumab Antibodies inhibit the binding of Vedolizumab to Human Integrin α4 β7.
Vedolizumab was coated at a concentration of 1 μg/mL, followed by the addition of His-tagged Human Integrin α4 β7 at 10 μg/mL and anti-Vedolizumab antibody that was serially diluted starting from 10 μg/mL. Detection was performed using THE™ His Tag Antibody [HRP], mAb, Mouse (GenScript, A00612) diluted at 1:5000.
MonoRab™ Anti-Vedolizumab Antibody (16G2), mAb, Rabbit (GenScript, A02326) exhibited non-inhibitory properties, MonoRab™ Anti-Vedolizumab Antibody (76H8), mAb, Rabbit (GenScript, A02325) showed weak inhibition, whereas MonoRab™ Anti-Vedolizumab Antibody (61A2), mAb, Rabbit (GenScript, A02324) demonstrated significant inhibitory properties with an IC₅₀ value of 0.57 µg/mL.

Standard Curve of Vedolizumab Sandwich ELISA by A02324 (61A2) and A02325 (76H8).
In this Sandwich ELISA assay, MonoRab™ Anti-Vedolizumab Antibody (61A2), mAb, Rabbit (GenScript, A02324) was coated at a concentration of 1 μg/ml, and MonoRab™ Anti-Vedolizumab Antibody (76H8), mAb, Rabbit (GenScript, A02325) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/mL. GenScript can provide customized conjugation service for this product per customer's request.
In this assay, a four-parameter logistic curve fitting program was used to create a standard curve with the R-Square equal to 0.99987. The typicaldynamic range of the assay is 0.03 - 2 ng/mL.

MRD Analysis of the Detection of Vedolizumab in Biological Matrix.
The MRD is the minimum dilution necessary for the detection of Vedolizumab 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 suggested that MRD was as 1:10.
In this ELISA assay, MonoRab™ Anti-Vedolizumab Antibody (61A2), mAb, Rabbit (GenScript, A02324) was coated at a concentration of 1 μg/ml, and MonoRab™ Anti-Vedolizumab Antibody (76H8), mAb, Rabbit (GenScript, A02325) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/mL.

Specificity analysis of the detection of Vedolizumab.

Vedolizumab standard samples at 7 concentrations were spiked with 10 µg/mL of human IgG1. The test result demonstrated that the high concentration of human IgG1 did not interfere with the detection of Vedolizumab.
In this ELISA assay, MonoRab™ Anti-Vedolizumab Antibody (61A2), mAb, Rabbit (GenScript, A02324) was coated at a concentration of 1 μg/ml, and MonoRab™ Anti-Vedolizumab Antibody (76H8), mAb, Rabbit (GenScript, A02325) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/mL.