Frontiers in immunology
A novel double-mAb sandwich ELISA utilizing broad-spectrum neutralizing antibodies as an in vitro alternative to the rabies vaccine NIH potency assay
By Cai M, Wu X, Shi L and 10 others.
Abstract
Background
Rabies remains a devastating zoonotic disease, with prevention measures relying heavily on post-exposure prophylaxis (PEP). Evaluating rabies vaccine potency traditionally relies on the in vivo National Institutes of Health (NIH) mouse challenge assay, which is limited by long testing cycles, low reproducibility, and high animal consumption. Developing broad-spectrum monoclonal antibodies (mAbs) and alternative in vitro assays is essential to advance rabies prevention and improve vaccine quality control.
Methods
In this study, we characterized three human-mouse chimeric mAbs (05-2F10, 06-2A12, and 07-4D8-1C1) with broad-spectrum neutralizing activity. Leveraging in vitro pseudovirus neutralization assays, we evaluated candidate mAbs for high neutralizing capacity and low sensitivity to variation to select optimal pairs for assay development. Using these selected mAbs, a double-antibody sandwich ELISA was established to quantify glycoprotein (G) antigen content in human rabies vaccines.