Human Anti-Abatacept Antibody bridging ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated overnight with Mouse Anti-Human CD152 Antibody, clone BNI3 (MCA1724) at a concentration of 1 µg/ml. After washing and blocking with PBST + 5% BSA, 10% human serum was added spiked with increasing concentrations of abatacept. Detection was performed using HRP conjugated Human Anti-Abatacept Antibody, clone AbD37058ia (HCA335) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements. HRP conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit (LNK002P).