Neutralizing antibodies are essential tools in antiviral therapy and epidemic preparedness,capable of directlyinhibiting viral entry and limiting disease progression.However,traditional antibody discovery strategies—...Neutralizing antibodies are essential tools in antiviral therapy and epidemic preparedness,capable of directlyinhibiting viral entry and limiting disease progression.However,traditional antibody discovery strategies—suchas animal immunization or B cell isolation from infected individuals—are often hindered by biosafety concerns,lengthy development timelines,and limited adaptability during outbreaks.In the present study,we aimed toestablish a robust and rapid in vitro platform for the efficient isolation of neutralizing antibodies targetingconserved viral epitopes.We developed an epitope-guided negative screening strategy that integrates phagedisplay technology with rational antigen mutagenesis to exclude antibodies against variable regions whileenriching for those that recognize functionally constrained epitopes.When applied to the receptor-binding domainof severe acute respiratory syndrome coronavirus 2,this method enabled the identification of six neutralizingantibodies(one IgG and five nanobodies)exhibiting broad-spectrum neutralizing activity across multiple viralvariants.Notably,antibodies recognizing distinct epitopes demonstrated significant synergistic neutralizationwhen used in combination(P<0.05).This screening approach facilitates the rapid discovery of potent andmutation-resistant antibodies and holds promise for application to other emerging pathogens.Our findingsunderscore the potential of epitope-guided,in vitro platforms in expediting therapeutic antibody developmentunder conditions of high biosafety requirements.展开更多
基金supported by the Natural Science Foundation of the Jiangsu Higher Education Institutions of China(Grant No.22KJB310001 to W.G.)the Special Project of the Jiangsu Provincial Department of Science and Technology(Grant No.BE2023603 to W.G.)the Nanjing Medical University Science and Technology Development Foundation(Grant No.NMUB20210006 to W.G.).
文摘Neutralizing antibodies are essential tools in antiviral therapy and epidemic preparedness,capable of directlyinhibiting viral entry and limiting disease progression.However,traditional antibody discovery strategies—suchas animal immunization or B cell isolation from infected individuals—are often hindered by biosafety concerns,lengthy development timelines,and limited adaptability during outbreaks.In the present study,we aimed toestablish a robust and rapid in vitro platform for the efficient isolation of neutralizing antibodies targetingconserved viral epitopes.We developed an epitope-guided negative screening strategy that integrates phagedisplay technology with rational antigen mutagenesis to exclude antibodies against variable regions whileenriching for those that recognize functionally constrained epitopes.When applied to the receptor-binding domainof severe acute respiratory syndrome coronavirus 2,this method enabled the identification of six neutralizingantibodies(one IgG and five nanobodies)exhibiting broad-spectrum neutralizing activity across multiple viralvariants.Notably,antibodies recognizing distinct epitopes demonstrated significant synergistic neutralizationwhen used in combination(P<0.05).This screening approach facilitates the rapid discovery of potent andmutation-resistant antibodies and holds promise for application to other emerging pathogens.Our findingsunderscore the potential of epitope-guided,in vitro platforms in expediting therapeutic antibody developmentunder conditions of high biosafety requirements.
