“Monoclonal antibodies (mAbs) are a rapidly advancing class of therapeutic glycoproteins that possess wide clinical utility owing to their biocompatibility, high antigen specificity, and targeted immune stimulation. These therapeutic properties depend greatly on the composition of the immunoglobulin G structure, both in terms of primary sequence and post-translational modifications; however, large-scale production in cell culture often results in heterogeneous mixtures that can profoundly affect clinical safety and efficacy. This places a high demand on analytical methods that afford comprehensive structural characterization of mAbs to ensure their stringent quality control.”
4238047
{:NJWQTL8H}
1
annual-reviews-author-date
50
default
1
title
45
https://proteoforms.org/wp-content/plugins/zotpress/
Recent Publications
4238047
{4238047:DMWWIRGI},{4238047:5NT4TR25},{4238047:P9H8UTZH},{4238047:ZHEDJI8R},{4238047:FVDV6MPG},{4238047:RT5ZJ5QL},{4238047:QLZER973}
1
annual-reviews-author-date
50
date
desc
1
title
45
https://proteoforms.org/wp-content/plugins/zotpress/
%7B%22status%22%3A%22success%22%2C%22updateneeded%22%3Afalse%2C%22instance%22%3Afalse%2C%22meta%22%3A%7B%22request_last%22%3A0%2C%22request_next%22%3A0%2C%22used_cache%22%3Atrue%7D%2C%22data%22%3A%5B%7B%22key%22%3A%22DMWWIRGI%22%2C%22library%22%3A%7B%22id%22%3A4238047%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Carfagno%20et%20al.%22%2C%22parsedDate%22%3A%222026-04-13%22%2C%22numChildren%22%3A1%7D%2C%22bib%22%3A%22%26lt%3Bdiv%20class%3D%26quot%3Bcsl-bib-body%26quot%3B%20style%3D%26quot%3Bline-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%26quot%3B%26gt%3B%5Cn%20%26lt%3Bdiv%20class%3D%26quot%3Bcsl-entry%26quot%3B%26gt%3BCarfagno%20AK%2C%20Lieu%20LB%2C%20Kline%20JT%2C%20Fornelli%20L%2C%20Durbin%20KR.%202026.%20%26lt%3Ba%20class%3D%26%23039%3Bzp-ItemURL%26%23039%3B%20target%3D%26%23039%3B_blank%26%23039%3B%20href%3D%26%23039%3Bhttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.analchem.5c06408%26%23039%3B%26gt%3BAutomating%20Middle-Down%20Mass%20Spectrometry%20Analysis%20for%20Extensive%20Antibody%20Characterization%26lt%3B%5C%2Fa%26gt%3B.%20%26lt%3Bi%26gt%3BAnal.%20Chem.%26lt%3B%5C%2Fi%26gt%3B%26lt%3B%5C%2Fdiv%26gt%3B%5Cn%26lt%3B%5C%2Fdiv%26gt%3B%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Automating%20Middle-Down%20Mass%20Spectrometry%20Analysis%20for%20Extensive%20Antibody%20Characterization%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Amy%20K.%22%2C%22lastName%22%3A%22Carfagno%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Linda%20B.%22%2C%22lastName%22%3A%22Lieu%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Jake%20T.%22%2C%22lastName%22%3A%22Kline%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Luca%22%2C%22lastName%22%3A%22Fornelli%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Kenneth%20R.%22%2C%22lastName%22%3A%22Durbin%22%7D%5D%2C%22abstractNote%22%3A%22Workflows%20and%20software%20tools%20for%20peptide%20mapping%20have%20been%20extensively%20optimized%20and%20simplified%20for%20widespread%20use.%20However%2C%20top-down%20and%20middle-down%20workflows%2C%20despite%20offering%20valuable%20proteoform%20information%20not%20obtainable%20by%20peptide%20mapping%2C%20remain%20demanding%20in%20terms%20of%20user%20expertise%20as%20well%20as%20instrument%20setup%2C%20data%20analysis%2C%20and%20result%20validation.%20Current%20software%20offerings%20have%20also%20not%20been%20tailored%20to%20address%20targeted%20protein%20characterization%20workflows%2C%20often%20necessitating%20multiple%20software%20tools%20to%20accomplish%20data%20analysis.%20To%20move%20toward%20routine%20intact%20protein%20characterization%20workflows%20with%20a%20single%20software%20tool%2C%20an%20automated%20platform%20named%20Proteoform%20Studio%20is%20introduced%20here.%20Proteoform%20Studio%20features%20a%20complete%20analysis%20pipeline%2C%20allowing%20a%20user%20to%20set%20up%20instrument%20runs%2C%20modify%20acquisition%20methods%2C%20rapidly%20deconvolute%20data%20to%20define%20intact%20mass%20features%2C%20search%20results%20to%20identify%20proteoforms%2C%20and%20obtain%20the%20highest%20quality%20proteoform%20characterization%20through%20automated%20aggregation%20of%20fragmentation%20results.%20Additionally%2C%20fragment%20ion%20matching%20settings%20for%20signal-to-noise%20ratio%20and%20isotope%20fit%20score%20were%20identified%20that%20produced%20automated%20matching%20results%20well-aligned%20with%20manual%20validation%20results%2C%20thereby%20reducing%20the%20need%20for%20time-consuming%20manual%20verification.%20With%20Proteoform%20Studio%20analysis%2C%20%26gt%3B80%25%20sequence%20coverage%20of%20light%20chain%20%28Lc%29%20and%20the%20N-terminal%20portion%20of%20heavy%20chain%20%28Fd%29%20antibody%20subunits%20was%20achieved%20in%20an%20automated%20fashion%20using%20a%20middle-down%20approach.%20These%20sequence%20coverage%20results%20utilized%20aggregation%20of%20spectral%20results%20from%20different%20fragmentation%20methods%20to%20take%20advantage%20of%20complementary%20fragment%20ion%20formation.%20Overall%2C%20introducing%20software%20to%20automate%20antibody%20subunit%20analysis%20will%20lower%20barriers%20to%20entry%20and%20eliminate%20data%20analysis%20bottlenecks%2C%20driving%20adoption%20in%20biopharmaceutical%20assays%20and%20ultimately%20leading%20to%20more%20precise%20characterization%20of%20therapeutic%20proteoform%20landscapes.%22%2C%22date%22%3A%222026-04-13%22%2C%22section%22%3A%22%22%2C%22partNumber%22%3A%22%22%2C%22partTitle%22%3A%22%22%2C%22DOI%22%3A%2210.1021%5C%2Facs.analchem.5c06408%22%2C%22citationKey%22%3A%22%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.analchem.5c06408%22%2C%22PMID%22%3A%22%22%2C%22PMCID%22%3A%22%22%2C%22ISSN%22%3A%220003-2700%22%2C%22language%22%3A%22%22%2C%22collections%22%3A%5B%5D%2C%22dateModified%22%3A%222026-04-22T15%3A05%3A55Z%22%7D%7D%2C%7B%22key%22%3A%22P9H8UTZH%22%2C%22library%22%3A%7B%22id%22%3A4238047%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Gould%20et%20al.%22%2C%22parsedDate%22%3A%222026-03-15%22%2C%22numChildren%22%3A0%7D%2C%22bib%22%3A%22%26lt%3Bdiv%20class%3D%26quot%3Bcsl-bib-body%26quot%3B%20style%3D%26quot%3Bline-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%26quot%3B%26gt%3B%5Cn%20%26lt%3Bdiv%20class%3D%26quot%3Bcsl-entry%26quot%3B%26gt%3BGould%20N%2C%20Johnson%20K%2C%20Marie%20A-L%2C%20Auclair%20JR%2C%20Ivanov%20AR.%202026.%20%26lt%3Ba%20class%3D%26%23039%3Bzp-ItemURL%26%23039%3B%20target%3D%26%23039%3B_blank%26%23039%3B%20href%3D%26%23039%3Bhttps%3A%5C%2F%5C%2Fwww.sciencedirect.com%5C%2Fscience%5C%2Farticle%5C%2Fpii%5C%2FS0731708525006508%26%23039%3B%26gt%3BCharge%20variant%20analysis%20of%20intact%20monoclonal%20antibody%20reference%20standards%20using%20microfluidic%20capillary%20zone%20electrophoresis%26lt%3B%5C%2Fa%26gt%3B.%20%26lt%3Bi%26gt%3BJournal%20of%20Pharmaceutical%20and%20Biomedical%20Analysis%26lt%3B%5C%2Fi%26gt%3B.%20270%3A117309%26lt%3B%5C%2Fdiv%26gt%3B%5Cn%26lt%3B%5C%2Fdiv%26gt%3B%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Charge%20variant%20analysis%20of%20intact%20monoclonal%20antibody%20reference%20standards%20using%20microfluidic%20capillary%20zone%20electrophoresis%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Noah%22%2C%22lastName%22%3A%22Gould%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Kendall%22%2C%22lastName%22%3A%22Johnson%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Anne-Lise%22%2C%22lastName%22%3A%22Marie%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Jared%20R.%22%2C%22lastName%22%3A%22Auclair%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Alexander%20R.%22%2C%22lastName%22%3A%22Ivanov%22%7D%5D%2C%22abstractNote%22%3A%22Monoclonal%20antibodies%20%28mAbs%29%20are%20among%20the%20most%20common%20biotherapeutics%2C%20and%20their%20clinical%20applications%20and%20global%20markets%20have%20demonstrated%20increased%20growth%20during%20the%20last%20decade%2C%20which%20are%20expected%20to%20grow%20further%20due%20to%20the%20shift%20of%20therapeutic%20treatments%20from%20general%20to%20personalized%20approaches.%20As%20the%20number%20and%20complexity%20of%20mAb-based%20therapeutics%20grow%2C%20there%20is%20an%20urgent%20need%20for%20highly%20efficient%20and%20high-throughput%20analytical%20techniques%20to%20characterize%20them%20and%20ensure%20their%20safety%2C%20potency%2C%20stability%2C%20and%20efficacy%20both%20from%20a%20development%20and%20regulatory%20standpoints.%20A%20key%20component%20in%20ensuring%20the%20methods%20used%20for%20the%20characterization%20of%20antibody-based%20therapeutics%20can%20provide%20accurate%20information%2C%20with%20the%20potential%20to%20influence%20important%20pipeline%20decisions%2C%20is%20the%20availability%20of%20diverse%20commercially%20available%20reference%20standards%20with%20well-known%20and%20catalogued%20properties%20that%20can%20serve%20as%20benchmarks%20and%20validation%20for%20the%20quality%20assurance%20of%20biotherapeutics%20in%20development.%20In%20this%20study%2C%20we%20developed%20and%20optimized%20a%20microfluidic%20capillary%20zone%20electrophoresis%20electrospray%20ionization-mass%20spectrometry%20%28CZE-ESI-MS%29%20method%2C%20utilizing%20the%20high-mass%20range%20MS%20data%20acquisition%2C%20to%20characterize%20three%20intact%20mAb%20reference%20standards%20and%20their%20proteoforms%20under%20near-native%20conditions%20using%20short%20analysis%20times%20%28%26lt%3B15%5Cu202fmin%29%2C%20low%20sample%20amounts%20%28500%20picograms%29%2C%20and%20minimal%20sample%20preparation.%20The%20developed%20CZE-MS%20method%20identified%20a%20number%20of%20modifications%20at%20the%20intact%20mAb%20level%2C%20including%20relatively%20high%20abundance%20C-terminal%20lysine%20clipping%20%28%26gt%3B95%5Cu202f%25%29%20and%20N-linked%20glycosylation%20%28%26gt%3B99%5Cu202f%25%29%2C%20as%20well%20as%20lower%20abundance%20modifications%2C%20such%20as%20deamidation%20%28%5Cu223c3%5Cu20137%5Cu202f%25%29%20and%20N-glycan%20sialylation%20%28%26lt%3B1.5%5Cu202f%25%29.%20The%20developed%20technique%20is%20uniquely%20suited%20for%20thorough%20and%20reproducible%20characterization%20of%20mAb-based%20biotherapeutics%20in%20industrial%20settings.%22%2C%22date%22%3A%222026-03-15%22%2C%22section%22%3A%22%22%2C%22partNumber%22%3A%22%22%2C%22partTitle%22%3A%22%22%2C%22DOI%22%3A%2210.1016%5C%2Fj.jpba.2025.117309%22%2C%22citationKey%22%3A%22%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fwww.sciencedirect.com%5C%2Fscience%5C%2Farticle%5C%2Fpii%5C%2FS0731708525006508%22%2C%22PMID%22%3A%22%22%2C%22PMCID%22%3A%22%22%2C%22ISSN%22%3A%220731-7085%22%2C%22language%22%3A%22%22%2C%22collections%22%3A%5B%5D%2C%22dateModified%22%3A%222025-12-10T15%3A58%3A12Z%22%7D%7D%2C%7B%22key%22%3A%225NT4TR25%22%2C%22library%22%3A%7B%22id%22%3A4238047%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Gould%20et%20al.%22%2C%22parsedDate%22%3A%222025-11-05%22%2C%22numChildren%22%3A0%7D%2C%22bib%22%3A%22%26lt%3Bdiv%20class%3D%26quot%3Bcsl-bib-body%26quot%3B%20style%3D%26quot%3Bline-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%26quot%3B%26gt%3B%5Cn%20%26lt%3Bdiv%20class%3D%26quot%3Bcsl-entry%26quot%3B%26gt%3BGould%20CE%2C%20Ma%20Q%2C%20Cha%20R%2C%20Frerichs%20VA%2C%20Friedman%20AE%2C%20et%20al.%202025.%20%26lt%3Ba%20class%3D%26%23039%3Bzp-ItemURL%26%23039%3B%20target%3D%26%23039%3B_blank%26%23039%3B%20href%3D%26%23039%3Bhttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Fjasms.5c00198%26%23039%3B%26gt%3BProteoform%20Characterization%20of%20HIV-1%20Broadly%20Neutralizing%20Antibody%20PGT%20121.414.LS%20Product%20Through%20Middle-Up%20and%20Bottom-Up%20Proteomics%20for%20Clinical%20Support%26lt%3B%5C%2Fa%26gt%3B.%20%26lt%3Bi%26gt%3BJ.%20Am.%20Soc.%20Mass%20Spectrom.%26lt%3B%5C%2Fi%26gt%3B%2036%2811%29%3A2445%5Cu201353%26lt%3B%5C%2Fdiv%26gt%3B%5Cn%26lt%3B%5C%2Fdiv%26gt%3B%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Proteoform%20Characterization%20of%20HIV-1%20Broadly%20Neutralizing%20Antibody%20PGT%20121.414.LS%20Product%20Through%20Middle-Up%20and%20Bottom-Up%20Proteomics%20for%20Clinical%20Support%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Connor%20E.%22%2C%22lastName%22%3A%22Gould%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Qing%22%2C%22lastName%22%3A%22Ma%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Raymond%22%2C%22lastName%22%3A%22Cha%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Valerie%20A.%22%2C%22lastName%22%3A%22Frerichs%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Alan%20E.%22%2C%22lastName%22%3A%22Friedman%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Robin%22%2C%22lastName%22%3A%22DiFrancesco%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Gene%20D.%22%2C%22lastName%22%3A%22Morse%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Troy%20D.%22%2C%22lastName%22%3A%22Wood%22%7D%5D%2C%22abstractNote%22%3A%22Broadly%20neutralizing%20antibody%20%28bNAb%29%20therapies%20are%20under%20development%20for%20the%20prevention%20and%20treatment%20of%20HIV-1%20infection%20as%20an%20alternative%20to%20conventional%20antiretroviral%20therapy%20because%20of%20their%20potential%20for%20sustained%20passive%20immunotherapy.%20Currently%2C%20bNAbs%20targeting%20the%20HIV-1%20viral%20envelope%20proteins%20are%20included%20in%20research%20protocols%20to%20evaluate%20their%20role%20in%20preventing%20transmission%20and%20in%20combination%20antiviral%20regimens%20to%20achieve%20viral%20suppression.%20While%20quantification%20of%20bNAbs%20is%20of%20clinical%20importance%2C%20validation%20of%20the%20structural%20fidelity%20of%20therapeutic%20antibodies%20is%20also%20an%20important%20consideration%20to%20assess%20the%20clinical%20efficacy.%20Here%2C%20we%20applied%20middle-up%20and%20bottom-up%20proteomics%20workflows%20for%20the%20quality%20assurance%20of%20the%20primary%20structure%20of%20the%20HIV-1%20bNAb%20PGT%20121.414.LS%20product.%20Middle-up%20and%20bottom-up%20proteomics%20workflows%20were%20performed%20using%20liquid%20chromatography%20coupled%20to%20an%20Orbitrap%20mass%20spectrometer.%20Middle-up%20analysis%20of%20the%20crystallizable%20fragment%20%28Fc%5C%2F2%29%2C%20light%20chain%20%28Lc%29%2C%20and%20fragment%20denaturation%20%28Fd%2C%20N-terminal%20fragment%20of%20the%20heavy%20chain%29%20regions%20of%20PGT%20121.414.LS%20was%20performed%20by%20using%20IdeS%20digestion%2C%20which%20revealed%20proteoform%20heterogeneity.%20To%20complement%20the%20middle-up%20approach%2C%20a%20bottom-up%20workflow%20combining%20trypsin%20and%20chymotrypsin%20digests%20was%20performed%20for%20detailed%20glycoform%20mapping%20and%20annotation.%20The%20bottom-up%20results%20indicated%20that%20Lc%20contained%20an%20additional%20N-terminal%20Ser%20residue.%20For%20the%20Fc%2C%20two%20abundant%20and%20two%20lower-abundance%20glycoforms%20of%20the%20heavy%20chain%20were%20detected%20that%20correspond%20to%20Asn-312%20%28Asn-297%20in%20the%20consensus%20heavy%20chain%20sequence%20of%20IgG%29.%20For%20Fd%2C%20bottom-up%20analysis%20revealed%20eight%20sialylated%20glycoforms%20and%20five%20nonsialylated%20glycoforms%20at%20Asn-124%20of%20the%20heavy%20chain.%20The%20results%20emphasize%20bNAb%20heterogeneity%2C%20which%20should%20be%20considered%20in%20affinity%20binding%20studies.%22%2C%22date%22%3A%222025-11-05%22%2C%22section%22%3A%22%22%2C%22partNumber%22%3A%22%22%2C%22partTitle%22%3A%22%22%2C%22DOI%22%3A%2210.1021%5C%2Fjasms.5c00198%22%2C%22citationKey%22%3A%22%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Fjasms.5c00198%22%2C%22PMID%22%3A%22%22%2C%22PMCID%22%3A%22%22%2C%22ISSN%22%3A%221044-0305%22%2C%22language%22%3A%22%22%2C%22collections%22%3A%5B%5D%2C%22dateModified%22%3A%222026-02-05T15%3A14%3A11Z%22%7D%7D%2C%7B%22key%22%3A%22FVDV6MPG%22%2C%22library%22%3A%7B%22id%22%3A4238047%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Benazza%20et%20al.%22%2C%22parsedDate%22%3A%222025-02-11%22%2C%22numChildren%22%3A0%7D%2C%22bib%22%3A%22%26lt%3Bdiv%20class%3D%26quot%3Bcsl-bib-body%26quot%3B%20style%3D%26quot%3Bline-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%26quot%3B%26gt%3B%5Cn%20%26lt%3Bdiv%20class%3D%26quot%3Bcsl-entry%26quot%3B%26gt%3BBenazza%20R%2C%20Letissier%20L%2C%20Papadakos%20G%2C%20Thom%20J%2C%20Diemer%20H%2C%20et%20al.%202025.%20%26lt%3Ba%20class%3D%26%23039%3Bzp-ItemURL%26%23039%3B%20target%3D%26%23039%3B_blank%26%23039%3B%20href%3D%26%23039%3Bhttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.analchem.4c03323%26%23039%3B%26gt%3BDevelopment%20of%20Top-Down%20Mass%20Spectrometry%20Strategies%20in%20the%20Chromatographic%20Time%20Scale%20%28LC-TD-MS%29%20for%20the%20Extended%20Characterization%20of%20an%20Anti-EGFR%20Single-Domain%20Antibody-Drug%20Conjugate%20in%20Both%20Reduced%20and%20Nonreduced%20Forms%26lt%3B%5C%2Fa%26gt%3B.%20%26lt%3Bi%26gt%3BAnal.%20Chem.%26lt%3B%5C%2Fi%26gt%3B%2097%285%29%3A2639%5Cu201347%26lt%3B%5C%2Fdiv%26gt%3B%5Cn%26lt%3B%5C%2Fdiv%26gt%3B%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Development%20of%20Top-Down%20Mass%20Spectrometry%20Strategies%20in%20the%20Chromatographic%20Time%20Scale%20%28LC-TD-MS%29%20for%20the%20Extended%20Characterization%20of%20an%20Anti-EGFR%20Single-Domain%20Antibody-Drug%20Conjugate%20in%20Both%20Reduced%20and%20Nonreduced%20Forms%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Rania%22%2C%22lastName%22%3A%22Benazza%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22L%5Cu00e9a%22%2C%22lastName%22%3A%22Letissier%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Greg%22%2C%22lastName%22%3A%22Papadakos%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Jen%22%2C%22lastName%22%3A%22Thom%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Helene%22%2C%22lastName%22%3A%22Diemer%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Graham%22%2C%22lastName%22%3A%22Cotton%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Sarah%22%2C%22lastName%22%3A%22Cianf%5Cu00e9rani%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Oscar%22%2C%22lastName%22%3A%22Hernandez-Alba%22%7D%5D%2C%22abstractNote%22%3A%22Even%20though%20mAbs%20have%20attracted%20the%20biggest%20interest%20in%20the%20development%20of%20therapeutic%20proteins%2C%20next-generation%20therapeutics%20such%20as%20single-domain%20antibodies%20%28sdAb%29%20are%20propelling%20increasing%20attention%20as%20new%20alternatives%20with%20appealing%20applications%20in%20different%20clinical%20areas.%20These%20constructs%20are%20small%20therapeutic%20proteins%20formed%20by%20a%20variable%20domain%20of%20the%20heavy%20chain%20of%20an%20antibody%20with%20multiple%20therapeutic%20and%20production%20benefits%20compared%20with%20their%20mAb%20counterparts.%20These%20proteins%20can%20be%20subjected%20to%20different%20bioconjugation%20processes%20to%20form%20single-domain%20antibody-drug%20conjugates%20%28sdADCs%29%20and%20hence%20increase%20their%20therapeutic%20potency%2C%20and%20akin%20to%20other%20therapeutic%20proteins%2C%20nanobodies%20and%20related%20products%20require%20dedicated%20analytical%20strategies%20to%20fully%20characterize%20their%20primary%20structure%20prior%20to%20their%20release%20to%20the%20market.%20In%20this%20study%2C%20we%20report%20for%20the%20first%20time%20the%20extensive%20sequence%20characterization%20of%20a%20conjugated%20anti-EGFR%2014%20kDa%20sdADC%20by%20using%20state-of-the-art%20top-down%20mass%20spectrometry%20strategies%20in%20combination%20with%20liquid%20chromatography%20%28LC-TD-MS%29.%20Mass%20analysis%20revealed%20a%20highly%20homogeneous%20sample%20with%20one%20conjugated%20molecule.%20Subsequently%2C%20the%20reduced%20sdADC%20was%20submitted%20to%20different%20fragmentation%20techniques%2C%20namely%2C%20higher-energy%20collisional%20dissociation%2C%20electron-transfer%20dissociation%2C%20and%20electron-transfer%20higher-energy%20collision%20dissociation%2C%20allowing%20to%20unambiguously%20assess%20the%20conjugation%20site%20with%2024%20diagnostic%20fragment%20ions%20and%2085%25%20of%20global%20sequence%20coverage.%20The%20sequence%20coverage%20of%20the%20nonreduced%20protein%20was%20significantly%20lower%20%28around%2016%25%29%3B%20however%2C%20the%20analysis%20of%20the%20fragmentation%20spectra%20corroborated%20the%20presence%20of%20the%20intramolecular%20disulfide%20bridge%20along%20with%20the%20localization%20of%20the%20conjugation%20site.%20Altogether%2C%20our%20results%20pinpoint%20the%20difficulties%20and%20challenges%20associated%20with%20the%20fragmentation%20of%20sdAb-derived%20formats%20in%20the%20LC%20time%20scale%20due%20to%20their%20remarkable%20stability%20as%20a%20consequence%20of%20the%20intramolecular%20disulfide%20bridge.%20However%2C%20the%20use%20of%20complementary%20activation%20techniques%20along%20with%20the%20identification%20of%20specific%20ion%20fragments%20allows%20an%20improved%20sequence%20coverage%2C%20the%20characterization%20of%20the%20intramolecular%20disulfide%20bond%2C%20and%20the%20unambiguous%20localization%20of%20the%20conjugation%20site.%22%2C%22date%22%3A%222025-02-11%22%2C%22section%22%3A%22%22%2C%22partNumber%22%3A%22%22%2C%22partTitle%22%3A%22%22%2C%22DOI%22%3A%2210.1021%5C%2Facs.analchem.4c03323%22%2C%22citationKey%22%3A%22%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.analchem.4c03323%22%2C%22PMID%22%3A%22%22%2C%22PMCID%22%3A%22%22%2C%22ISSN%22%3A%220003-2700%22%2C%22language%22%3A%22%22%2C%22collections%22%3A%5B%5D%2C%22dateModified%22%3A%222025-02-12T17%3A47%3A58Z%22%7D%7D%2C%7B%22key%22%3A%22ZHEDJI8R%22%2C%22library%22%3A%7B%22id%22%3A4238047%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Schairer%20et%20al.%22%2C%22parsedDate%22%3A%222025-01-01%22%2C%22numChildren%22%3A0%7D%2C%22bib%22%3A%22%26lt%3Bdiv%20class%3D%26quot%3Bcsl-bib-body%26quot%3B%20style%3D%26quot%3Bline-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%26quot%3B%26gt%3B%5Cn%20%26lt%3Bdiv%20class%3D%26quot%3Bcsl-entry%26quot%3B%26gt%3BSchairer%20J%2C%20R%5Cu00f6mer%20J%2C%20Neus%5Cu00fc%5Cu00df%20C.%202025.%20%26lt%3Ba%20class%3D%26%23039%3Bzp-ItemURL%26%23039%3B%20target%3D%26%23039%3B_blank%26%23039%3B%20href%3D%26%23039%3Bhttps%3A%5C%2F%5C%2Fwww.sciencedirect.com%5C%2Fscience%5C%2Farticle%5C%2Fpii%5C%2FS0731708524005375%26%23039%3B%26gt%3BCE-MS%20and%20CE-MS%5C%2FMS%20for%20the%20multiattribute%20analysis%20of%20monoclonal%20antibody%20variants%20at%20the%20subunit%20level%26lt%3B%5C%2Fa%26gt%3B.%20%26lt%3Bi%26gt%3BJournal%20of%20Pharmaceutical%20and%20Biomedical%20Analysis%26lt%3B%5C%2Fi%26gt%3B.%20252%3A116495%26lt%3B%5C%2Fdiv%26gt%3B%5Cn%26lt%3B%5C%2Fdiv%26gt%3B%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22CE-MS%20and%20CE-MS%5C%2FMS%20for%20the%20multiattribute%20analysis%20of%20monoclonal%20antibody%20variants%20at%20the%20subunit%20level%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Jasmin%22%2C%22lastName%22%3A%22Schairer%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Jennifer%22%2C%22lastName%22%3A%22R%5Cu00f6mer%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Christian%22%2C%22lastName%22%3A%22Neus%5Cu00fc%5Cu00df%22%7D%5D%2C%22abstractNote%22%3A%22The%20analysis%20of%20product-related%20substances%20and%20impurities%20is%20a%20critical%20step%20in%20the%20biopharmaceutical%20quality%20control%20of%20multiattribute%20monoclonal%20antibodies%20%28mAbs%29%2C%20as%20posttranslational%20modifications%20or%20other%20variants%20can%20influence%20the%20product%26%23039%3Bs%20biological%20activity.%20Many%20approaches%20are%20available%20for%20variant%20analysis%3B%20however%2C%20they%20are%20either%20variant-specific%2C%20mAb-specific%2C%20time-consuming%2C%20or%20require%20expensive%20equipment.%20Here%2C%20we%20present%20a%20generic%20capillary%20electrophoretic%20method%20based%20on%20a%20neutral-coated%20capillary%20which%20was%20coupled%20to%20mass%20spectrometry%20%28MS%29%20via%20the%20nanoCEasy%20interface%20for%20mAb%20variant%20analysis%20at%20the%20subunit%20level%20%28enzymatically%20digested%20and%20reduced%20mAb%29.%20The%20method%20enabled%20the%20separation%20of%20several%20%28i%29%20size%20variants%20%28e.g.%20glycosylation%20variants%29%20and%20%28ii%29%20charge%20variants%20%28e.g.%20c-terminal%20lysin%20clipping%29%20as%20well%20as%20%28iii%29%20multiple%20other%20proteoforms%20%28e.g.%20additional%20glycation%29%20and%20%28iv%29%20incompletely%20reduced%20subunits.%20Separated%20variants%20were%20confirmed%20by%20MS%5C%2FMS%20fragmentation%20even%20for%20small%20mass%20deviations%20like%20deamidation%20or%20open%20disulfide%20bridges.%20The%20system%2C%20initially%20developed%20for%20one%20mAb%2C%20was%20tested%20with%20nine%20other%20IgG1s%20to%20show%20the%20general%20applicability%20of%20the%20system.%20The%20presented%20multiattribute%20method%20enables%20fast%20and%20detailed%20characterization%20of%20mAb%20variants%20with%20little%20sample%20preparation%20and%20relatively%20simple%20separation%20equipment%20enabling%20the%20separation%20of%20a%20large%20set%20of%20mAb%20variants.%22%2C%22date%22%3A%222025-01-01%22%2C%22section%22%3A%22%22%2C%22partNumber%22%3A%22%22%2C%22partTitle%22%3A%22%22%2C%22DOI%22%3A%2210.1016%5C%2Fj.jpba.2024.116495%22%2C%22citationKey%22%3A%22%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fwww.sciencedirect.com%5C%2Fscience%5C%2Farticle%5C%2Fpii%5C%2FS0731708524005375%22%2C%22PMID%22%3A%22%22%2C%22PMCID%22%3A%22%22%2C%22ISSN%22%3A%220731-7085%22%2C%22language%22%3A%22%22%2C%22collections%22%3A%5B%5D%2C%22dateModified%22%3A%222025-02-12T17%3A49%3A54Z%22%7D%7D%2C%7B%22key%22%3A%22QLZER973%22%2C%22library%22%3A%7B%22id%22%3A4238047%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Lieu%20et%20al.%22%2C%22parsedDate%22%3A%222024-11-19%22%2C%22numChildren%22%3A0%7D%2C%22bib%22%3A%22%26lt%3Bdiv%20class%3D%26quot%3Bcsl-bib-body%26quot%3B%20style%3D%26quot%3Bline-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%26quot%3B%26gt%3B%5Cn%20%26lt%3Bdiv%20class%3D%26quot%3Bcsl-entry%26quot%3B%26gt%3BLieu%20LB%2C%20Nagy%20C%2C%20Huang%20J%2C%20Mullen%20C%2C%20McAlister%20GC%2C%20et%20al.%202024.%20%26lt%3Ba%20class%3D%26%23039%3Bzp-ItemURL%26%23039%3B%20target%3D%26%23039%3B_blank%26%23039%3B%20href%3D%26%23039%3Bhttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.analchem.4c03872%26%23039%3B%26gt%3BEnhanced%20Payload%20Localization%20in%20Antibody%5Cu2013Drug%20Conjugates%20Using%20a%20Middle-Down%20Mass%20Spectrometry%20Approach%20with%20Proton%20Transfer%20Charge%20Reduction%26lt%3B%5C%2Fa%26gt%3B.%20%26lt%3Bi%26gt%3BAnal.%20Chem.%26lt%3B%5C%2Fi%26gt%3B%2096%2846%29%3A18483%5Cu201390%26lt%3B%5C%2Fdiv%26gt%3B%5Cn%26lt%3B%5C%2Fdiv%26gt%3B%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Enhanced%20Payload%20Localization%20in%20Antibody%5Cu2013Drug%20Conjugates%20Using%20a%20Middle-Down%20Mass%20Spectrometry%20Approach%20with%20Proton%20Transfer%20Charge%20Reduction%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Linda%20B.%22%2C%22lastName%22%3A%22Lieu%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Cynthia%22%2C%22lastName%22%3A%22Nagy%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Jingjing%22%2C%22lastName%22%3A%22Huang%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Christopher%22%2C%22lastName%22%3A%22Mullen%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Graeme%20C.%22%2C%22lastName%22%3A%22McAlister%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Vlad%22%2C%22lastName%22%3A%22Zabrouskov%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Kristina%22%2C%22lastName%22%3A%22Srzenti%5Cu0107%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Kenneth%20R.%22%2C%22lastName%22%3A%22Durbin%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Rafael%20D.%22%2C%22lastName%22%3A%22Melani%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Luca%22%2C%22lastName%22%3A%22Fornelli%22%7D%5D%2C%22abstractNote%22%3A%22Antibody%5Cu2013drug%20conjugates%20%28ADCs%29%20represent%20a%20novel%20class%20of%20immunoconjugates%20with%20growing%20therapeutic%20relevance%2C%20since%20they%20combine%20the%20efficacy%20of%20cytotoxic%20drugs%20with%20the%20specificity%20of%20antibodies.%20However%2C%20by%20design%2C%20ADCs%20introduce%20structural%20features%20into%20the%20monoclonal%20antibody%20scaffold%20that%20complicate%20their%20analysis.%20Payload%20attachment%20to%20cysteine%20or%20lysine%20residues%20can%20often%20result%20in%20product%20heterogeneity%2C%20regarding%20both%20the%20number%20of%20attached%20drug%20molecules%20and%20their%20conjugation%20site%2C%20necessitating%20the%20use%20of%20state-of-the-art%20MS%20instrumentation%20to%20elucidate%20their%20complexity.%20In%20middle-down%20mass%20spectrometry%20%28MD%20MS%29%2C%20the%20gas-phase%20sequencing%20of%20%5Cu223c25%20kDa%20ADC%20subunits%20with%20different%20ion%20activation%20techniques%20generally%20produces%20rich%20fragmentation%20mass%20spectra%3B%20however%2C%20spectral%20congestion%20can%20cause%20some%20fragment%20ions%20to%20go%20undetected%2C%20including%20those%20that%20can%20pinpoint%20the%20exact%20location%20of%20payload%20conjugation%20sites.%20Proton%20transfer%20charge%20reduction%20%28PTCR%29%20can%20substantially%20simplify%20fragment%20ion%20spectra%2C%20thereby%20unveiling%20the%20presence%20of%20product%20ions%20whose%20signals%20were%20previously%20suppressed.%20Herein%2C%20we%20present%20an%20MD%20MS%20strategy%20relying%20on%20the%20use%20of%20PTCR%20to%20investigate%20a%20cysteine-based%20ADC%20mimic%20with%20a%20variable%20drug-to-antibody%20ratio%2C%20targeting%20the%20unambiguous%20localization%20of%20payload%20conjugation%20sites.%20Unlike%20traditional%20tandem%20MS%20experiments%20%28MS2%29%2C%20which%20could%20not%20provide%20a%20complete%20map%20of%20conjugation%20sites%2C%20a%20single%20PTCR-based%20experiment%20%28MS3%29%20proved%20to%20be%20sufficient%20to%20achieve%20this%20goal%20across%20all%20variably%20modified%20ADC%20subunits%2C%20including%20isomeric%20ones.%20Combining%20the%20results%20obtained%20from%20orthogonal%20ion%20activation%20techniques%20followed%20by%20PTCR%20further%20strengthened%20the%20confidence%20in%20the%20assignments.%22%2C%22date%22%3A%222024-11-19%22%2C%22section%22%3A%22%22%2C%22partNumber%22%3A%22%22%2C%22partTitle%22%3A%22%22%2C%22DOI%22%3A%2210.1021%5C%2Facs.analchem.4c03872%22%2C%22citationKey%22%3A%22%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.analchem.4c03872%22%2C%22PMID%22%3A%22%22%2C%22PMCID%22%3A%22%22%2C%22ISSN%22%3A%220003-2700%22%2C%22language%22%3A%22%22%2C%22collections%22%3A%5B%5D%2C%22dateModified%22%3A%222024-11-22T15%3A29%3A46Z%22%7D%7D%2C%7B%22key%22%3A%22RT5ZJ5QL%22%2C%22library%22%3A%7B%22id%22%3A4238047%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Liu%20et%20al.%22%2C%22parsedDate%22%3A%222024-11-19%22%2C%22numChildren%22%3A0%7D%2C%22bib%22%3A%22%26lt%3Bdiv%20class%3D%26quot%3Bcsl-bib-body%26quot%3B%20style%3D%26quot%3Bline-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%26quot%3B%26gt%3B%5Cn%20%26lt%3Bdiv%20class%3D%26quot%3Bcsl-entry%26quot%3B%26gt%3BLiu%20J%2C%20Liu%20Z%2C%20Zhao%20H%2C%20Xiao%20C%2C%20Yang%20X%2C%20Wang%20F.%202024.%20%26lt%3Ba%20class%3D%26%23039%3Bzp-ItemURL%26%23039%3B%20target%3D%26%23039%3B_blank%26%23039%3B%20href%3D%26%23039%3Bhttps%3A%5C%2F%5C%2Fwww.sciencedirect.com%5C%2Fscience%5C%2Farticle%5C%2Fpii%5C%2FS0003267024012510%26%23039%3B%26gt%3BSequencing%20the%20Monoclonal%20Antibody%20Variable%20Regions%20Using%20Multiple%20Charge%20Integration%20Middle-Down%20Strategy%20and%20Ultraviolet%20Photodissociation%26lt%3B%5C%2Fa%26gt%3B.%20%26lt%3Bi%26gt%3BAnalytica%20Chimica%20Acta%26lt%3B%5C%2Fi%26gt%3B.%20343450%26lt%3B%5C%2Fdiv%26gt%3B%5Cn%26lt%3B%5C%2Fdiv%26gt%3B%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Sequencing%20the%20Monoclonal%20Antibody%20Variable%20Regions%20Using%20Multiple%20Charge%20Integration%20Middle-Down%20Strategy%20and%20Ultraviolet%20Photodissociation%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Jialiang%22%2C%22lastName%22%3A%22Liu%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Zheyi%22%2C%22lastName%22%3A%22Liu%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Heng%22%2C%22lastName%22%3A%22Zhao%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Chunlei%22%2C%22lastName%22%3A%22Xiao%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Xueming%22%2C%22lastName%22%3A%22Yang%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Fangjun%22%2C%22lastName%22%3A%22Wang%22%7D%5D%2C%22abstractNote%22%3A%22Background%5CnTherapeutic%20monoclonal%20antibodies%20%28mAbs%29%20have%20become%20essential%20biopharmaceuticals%20for%20clinical%20targeted%20therapies%20due%20to%20their%20high%20specificity%2C%20affinity%20and%20low%20side%20effects.%20The%20specificity%20and%20affinity%20of%20mAb%20to%20targeting%20antigen%20are%20mainly%20dependent%20on%20the%20three%20complementarity%20determining%20regions%20%28CDRs%29%20with%20high%20variations%20in%20amino%20acid%20sequences.%20Therefore%2C%20mAb%20CDR%20sequencing%20is%20crucial%20for%20the%20characterization%20of%20therapeutic%20mAbs.%20Here%2C%20we%20developed%20a%20193-nm%20ultraviolet%20photodissociation%20%28UVPD%29%20based%20multiple%20charge%20integration%20middle-down%20mass%20spectrometry%20%28MCI-MDMS%29%20strategy%20for%20mAb%20sequencing.%5CnResults%5CnWe%20demonstrate%20that%20the%20UVPD%20spectra%20of%20mAb%20subunit%20ions%20with%20different%20charge%20states%20exhibit%20high%20complementarity%2C%20and%20integration%20can%20result%20in%20higher%20sequence%20coverage%20compared%20to%20single%20charge%20states.%20Finally%2C%20over%2095%25%20sequence%20coverage%20of%20two%20different%20mAbs%20has%20been%20achieved%20with%20full%20sequence%20coverage%20of%20CDRs%2C%20underscoring%20the%20great%20potential%20of%20this%20strategy%20in%20accurate%20sequencing%20of%20mAb%20variable%20regions.%20Compared%20with%20the%20conventional%20higher%20energy%20collisional%20dissociation%20%28HCD%29%20strategy%20of%20mAb%20subunit%20sequencing%2C%20the%20sequence%20coverage%20of%20CDRs%20at%20single%20UVPD%20subunit%20charge%20state%20has%20increased%20by%20an%20average%20of%2030%25.%20In%20addition%2C%20almost%20complete%20sequence%20coverage%20of%20mAb%20ensures%20the%20accurate%20localization%20of%20mAb%20post-translational%20modifications%20%28PTMs%29%2C%20including%20glycosylation%20of%20two%20different%20sites%2C%20C-terminal%20lysine%20truncation%2C%20and%20N-terminal%20cyclization%20of%20glutamine.%5CnSignificance%5CnThe%20integration%20of%20MCI-MDMS%20and%20UVPD%20realizes%20high%20sequence%20coverage%20and%20reliable%20PTM%20determination%20of%20mAbs.%20This%20integrated%20strategy%20holds%20significant%20promise%20for%20accurate%20analysis%20of%20antibody-drug%20conjugates%2C%20polyclonal%20antibodies%20and%20unknown%20mAbs%20including%20sequences%20and%20PTMs%2C%20and%20providing%20a%20crucial%20tool%20for%20the%20discovery%20and%20development%20of%20therapeutic%20mAbs.%22%2C%22date%22%3A%222024-11-19%22%2C%22section%22%3A%22%22%2C%22partNumber%22%3A%22%22%2C%22partTitle%22%3A%22%22%2C%22DOI%22%3A%2210.1016%5C%2Fj.aca.2024.343450%22%2C%22citationKey%22%3A%22%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fwww.sciencedirect.com%5C%2Fscience%5C%2Farticle%5C%2Fpii%5C%2FS0003267024012510%22%2C%22PMID%22%3A%22%22%2C%22PMCID%22%3A%22%22%2C%22ISSN%22%3A%220003-2670%22%2C%22language%22%3A%22%22%2C%22collections%22%3A%5B%5D%2C%22dateModified%22%3A%222024-11-22T20%3A32%3A29Z%22%7D%7D%5D%7D
Carfagno AK, Lieu LB, Kline JT, Fornelli L, Durbin KR. 2026. Automating Middle-Down Mass Spectrometry Analysis for Extensive Antibody Characterization. Anal. Chem.
Gould N, Johnson K, Marie A-L, Auclair JR, Ivanov AR. 2026. Charge variant analysis of intact monoclonal antibody reference standards using microfluidic capillary zone electrophoresis. Journal of Pharmaceutical and Biomedical Analysis. 270:117309
Gould CE, Ma Q, Cha R, Frerichs VA, Friedman AE, et al. 2025. Proteoform Characterization of HIV-1 Broadly Neutralizing Antibody PGT 121.414.LS Product Through Middle-Up and Bottom-Up Proteomics for Clinical Support. J. Am. Soc. Mass Spectrom. 36(11):2445–53
Benazza R, Letissier L, Papadakos G, Thom J, Diemer H, et al. 2025. Development of Top-Down Mass Spectrometry Strategies in the Chromatographic Time Scale (LC-TD-MS) for the Extended Characterization of an Anti-EGFR Single-Domain Antibody-Drug Conjugate in Both Reduced and Nonreduced Forms. Anal. Chem. 97(5):2639–47
Schairer J, Römer J, Neusüß C. 2025. CE-MS and CE-MS/MS for the multiattribute analysis of monoclonal antibody variants at the subunit level. Journal of Pharmaceutical and Biomedical Analysis. 252:116495
Lieu LB, Nagy C, Huang J, Mullen C, McAlister GC, et al. 2024. Enhanced Payload Localization in Antibody–Drug Conjugates Using a Middle-Down Mass Spectrometry Approach with Proton Transfer Charge Reduction. Anal. Chem. 96(46):18483–90
Liu J, Liu Z, Zhao H, Xiao C, Yang X, Wang F. 2024. Sequencing the Monoclonal Antibody Variable Regions Using Multiple Charge Integration Middle-Down Strategy and Ultraviolet Photodissociation. Analytica Chimica Acta. 343450