FigsharePub Date : 2023-09-08DOI: 10.1021/jasms.3c00215.s001
Rui Zhao (43391), Ning Liu (168279), Zhen Zheng (445428), Gongyu Li (2542105)
{"title":"Enhanced Stability Differentiation of TherapeuticnPolyclonal Antibodies with All Ion Unfolding-Ion Mobility-Mass Spectrometry","authors":"Rui Zhao (43391), Ning Liu (168279), Zhen Zheng (445428), Gongyu Li (2542105)","doi":"10.1021/jasms.3c00215.s001","DOIUrl":"https://doi.org/10.1021/jasms.3c00215.s001","url":null,"abstract":"Compared with monoclonalnantibodies, polyclonal antibodies (pAbs)nhave rather significant characteristics, including lower cost, shorternproduction cycle, and higher affinity. Therefore, to facilitate theirnapplications in clinic, it is equally critical to comprehensivelyncharacterize the conformational stabilities of pAb at the molecularnweight-resolved scale, which is technically challenging due to thenlack of an effective analytical tool capable of simultaneously providingnboth stability and molecular weight information within an acceptablenerror range. Ion mobility-mass spectrometry (IM-MS) has grown as annalternative to rapidly assess protein conformational stability withnaccurate molecular weight information maintained, especially whennequipped with a collision-induced unfolding (CIU) regime. Dynamicnand transient conformational intermediates can be captured with thenCIU-IM-MS technique, adding to traditional static structural measurementsnwith collisional cross section. Most CIU-IM-MS-centered protocolsnare focusing on the application to isolated, targeted protein ions,nnamely, analyzing one single charge state at one time, limiting itsnanalytical throughput and speed. In this study, we employed an enhancednunfolding regime, all ion unfolding (AIU), capable of the simultaneousnoperation of numerous ions at a time during stepped unfolding processesnto analyze pAb. Results show that AIU can quantitatively characterizenthe subtle differences in conformational stability among four structurallynsimilar pAbs with improved resolving capability by around a 2–4-foldnincrement in both stability and structure differentiating parameters.nBesides, AIU also benefits from considerably saved time cost and improvednspectrum quality with an elevated signal-to-noise ratio.","PeriodicalId":523787,"journal":{"name":"Figshare","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147921945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}