{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T06:49:36Z","timestamp":1791355776531,"version":"4.3.1"},"reference-count":26,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Rabies remains a devastating zoonotic disease, with prevention measures relying heavily on post-exposure prophylaxis (PEP). Evaluating rabies vaccine potency traditionally relies on the\n                      <jats:italic>in vivo<\/jats:italic>\n                      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\n                      <jats:italic>in vitro<\/jats:italic>\n                      assays is essential to advance rabies prevention and improve vaccine quality control.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      In this study, we characterized three human-mouse chimeric mAbs (05-2F10, 06-2A12, and 07-4D8-1C1) with broad-spectrum neutralizing activity. Leveraging\n                      <jats:italic>in vitro<\/jats:italic>\n                      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.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The three chimeric mAbs exhibited exceptional\n                      <jats:italic>in vitro<\/jats:italic>\n                      neutralization potency and breadth. The established double-antibody sandwich ELISA successfully quantified G antigen content in human rabies vaccines. Importantly, a high correlation was demonstrated between the antigen content quantified by this ELISA and the potency data generated via the traditional\n                      <jats:italic>in vivo<\/jats:italic>\n                      NIH mouse challenge test.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Collectively, these findings provide a robust technical framework for replacing the traditional NIH potency assay with a reliable\n                      <jats:italic>in vitro<\/jats:italic>\n                      method, fully aligning with the 3Rs principles of animal welfare.\n                    <\/jats:p>\n                  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