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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