Biolayer interferometry predicts ELISA performance of monoclonal antibody pairs for Plasmodium falciparum histidine-rich protein 2

•Kinetic parameters of 15 anti-HRP2 antibodies are measured by biolayer interferometry.•Kinetic constants are compared to a checkerboard ELISA of 225 antibody pairs.•Biolayer interferometry predicts antibody pair performance for HRP2 ELISA.•Capture mAbs with low koff and detection mAbs with high kon are best in HRP2 ELISA.

Predicting antibody pair performance in a sandwich format streamlines development of antibody-based diagnostics and laboratory research tools, such as enzyme-linked immunosorbent assays (ELISAs) and lateral flow immunoassays (LFAs). We have evaluated panels of monoclonal antibodies against the malarial parasite biomarker Plasmodium falciparum histidine rich protein 2 (HRP2), including 9 new monoclonal antibodies, using biolayer interferometry (BLI) and screened antibody pairs in a checkerboard ELISA. This study showed BLI predicts antibody pair ELISA performance for HRP2. Pairs that included capture antibodies with low off-rate constants and detection antibodies with high on-rate constants performed best in an ELISA format.

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