mAbs最新文献

{"title":"2D NMR assessment of structural consistency and functional relationships in monoclonal antibodies.","authors":"Lingfei Wang, M Jasmin Donohue, Blaine Ballard, Kimberly Nadendla, Ken Skidmore, Aaron T Wecksler","doi":"10.1080/19420862.2026.2672774","DOIUrl":"10.1080/19420862.2026.2672774","url":null,"abstract":"<p><p>Two-dimensional (2D) NMR at natural-abundance ¹³C has emerged as a powerful technique for generating high-resolution structural fingerprints of monoclonal antibody (mAb) higher-order structure. While increasingly applied in structural consistency and comparability assessments, the fraction of a heterogeneous mAb population required to produce a detectable change in the NMR fingerprint - and how this relates to functional impact - has not been systematically defined. In this study, we investigated the relationship between population-level structural changes detected by 2D NMR and functional consequences using antibody-dependent cellular cytotoxicity (ADCC) as a readout. A series of co-mixed mAb samples containing increasing levels of aglycosylation was prepared using homodimeric non-glycosylated heavy chain as a surrogate for aglycosylated species. Changes in 2D NMR fingerprints were evaluated using ECHOS correlation and principal component analysis and compared with FcγRIIIa binding and ADCC activity. Detectable changes in the 2D NMR fingerprint were observed only when aglycosylated mAb comprised approximately 25% or more of the population. In contrast, significant reductions in Fc receptor binding and ADCC activity occurred at lower aglycosylated fractions (approximately 10-20%). These findings demonstrate that population-level structural perturbations must exceed functional sensitivity thresholds to be detected by 2D NMR. Collectively, this work provides important context for interpreting this type of 2D NMR data in structural consistency and biosimilarity assessments, highlighting both the strengths and interpretive limits of the technique when low-level heterogeneity is present in mAb populations.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2672774"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13178170/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147941968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Using mass spectrometry for robust identity release testing in a global quality control network.","authors":"Sebastien Leibe, Sebastien Violini, Moritz Vollrath, Anja Bathke, Hong Nguyen, Harry Yu, Jack Harris, Christopher Yu, Jan O Stracke, Hilary Pallat, Christian Bell, Elizabeth J Johnson, Cinzia Stella, Patrick Bulau","doi":"10.1080/19420862.2026.2643964","DOIUrl":"10.1080/19420862.2026.2643964","url":null,"abstract":"<p><p>Product identity (ID) testing is a fundamental requirement in pharmaceutical quality control. Roche/Genentech has developed and established a mass spectrometry-based platform ID method for routine release testing of biologics in the global clinical and commercial quality control environment. The established method, using peptide mapping combined with liquid chromatography-mass spectrometry, allows automated data evaluation. Previous experience shows that this approach is highly user-friendly in regulated testing environments, significantly reduces validation work for new products, and, due to its high robustness, results in a reduced error rate during test execution and data evaluation. Due to the specific analysis of product peptides, the method also allows for a sensitive and high-resolution evaluation of further product variants that are attributable to chemical and/or post-translational amino acid modifications. The data already recorded for identity testing purposes can therefore be immediately reevaluated during a quantitative product quality impact assessment following unexpected events in the manufacturing process, thus saving additional analyses during root cause analysis and future failure mitigation. In summary, this technology, established and qualified at all Roche/Genentech quality control sites, enables improved product monitoring beyond the routine release and stability testing methods used.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2643964"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12977250/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147434056","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preclinical characterization of AZD1163, a first-in-class anti-PAD2/4 bispecific antibody for the treatment of rheumatoid arthritis.","authors":"Gary P Sims, Lacie Scaletta, John Andrews, Dorothy A Sims, Martin Strain, Anna Sigurdardottir, Teneema Kuriakose, Elizabeth England, Lichchavi Rajasinghe, Fanyi Jiang, Ximing Xiong, Frances Neal, Lisa Vinall, Philip Newton, Elin Boger, Chong Kim, Helena Dahlbäck, Susanne Prothon, Jacob Leander, Erika Darrah, Mia Collins, Garry Douglas, Nicholas White, Katie Day, Katherine A Vousden, Catherine E Huntington, David Close","doi":"10.1080/19420862.2026.2657629","DOIUrl":"10.1080/19420862.2026.2657629","url":null,"abstract":"<p><p>Anti-citrullinated protein autoantibodies promote inflammation and joint tissue injury and define a poor prognostic group of patients with rheumatoid arthritis (RA). Citrullinated autoantigens that drive this autoimmune response are generated by peptidylarginine deiminase (PAD) enzymes, which are predominately expressed and released by neutrophils and monocytes. Accordingly, blocking the enzymatic activity of PADs to curb the generation of citrullinated autoantigens that drive autoimmunity and tissue injury may provide therapeutic benefit. Herein, we developed a high affinity, bispecific, anti-PAD2/4, effector-null antibody, AZD1163, which potently inhibits recombinant PAD2 and PAD4 activity in both histone H3 and fibrinogen citrullination assays. AZD1163 inhibited all endogenous PAD activity in the serum of patients with RA irrespective of the presence of anti-PAD4 autoantibodies, and neutralized PAD activity in synovial fluid. AZD1163 also bound and internalized PADs expressed on cell membranes into low pH endosomes for degradation, reducing the surface expression and catalytic potential. Binding of AZD1163 to neutrophils and monocytes did not trigger complement-dependent cytotoxicity, antibody-dependent cellular cytotoxicity, or the production of proinflammatory cytokines, or otherwise impact neutrophil phagocytosis, production of reactive oxygen species, or NETosis. In a non-human primate study of pharmacokinetics (PK) and pharmacodynamics, a single dose of AZD1163 exhibited PK consistent with a half-life extended antibody and a rapid and durable suppression of endogenous PAD activity. AZD1163 has a favorable preclinical safety profile. Collectively, these in vitro and in vivo pharmacology and safety data support the clinical development of AZD1163 as a novel therapeutic strategy for RA by reducing autoantigen load.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2657629"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13085943/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147690958","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"AlphaFold 3 prediction of TCR mimic Fv - pMHC complex structures and their validation and optimization.","authors":"Abhijit Sarkar, Sara Michael, Zening Wang, Xin Ge","doi":"10.1080/19420862.2026.2719263","DOIUrl":"10.1080/19420862.2026.2719263","url":null,"abstract":"<p><p>Monoclonal antibody-derived chimeric antigen receptors (CAR) that mimic T‑cell receptors (TCR) on binding to peptide-major histocompatibility complex (pMHC) and activating T‑cell functions hold great promise for the development of effective immunotherapy. This study applied AlphaFold 3 (AF3) to model the quaternary structures of TCR mimic antibody variable fragment (TCRm Fv) complexed with NY-ESO-1_157-165/HLA-A*02:01. Benchmark study suggested that reliable TCRm Fv-pMHC structures were achieved by AF3 with high confidence. Toward NY-ESO-1/A2-specific CAR clones, AF3 prediction revealed their intermolecular geometry resembling the canonical TCR engagement, and binding avidity tests confirmed the peptide-dependent recognition of pMHC. Molecular interactions between Fv and the peptide antigen and its binding groove on MHC were further pinpointed, and the predicted epitopes and paratopes were validated by mutagenesis studies. Aiming to generate TCRm Fv mutants of improved affinity, AF3 aided designs with larger Fv-pHLA contact surface areas and more hydrogen bonds toward the peptide antigen were successfully identified. However, these AF3‑designed mutants failed to deliver enhancement on binding avidity or functional potency in experimental tests. Overall, AF3 is a highly valuable tool for TCRm Fv-pMHC complex modeling, and its combination with force field-based methods will be desirable to aid optimization tasks.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2719263"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13502005/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148794990","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The making of multispecific immunoglobulins - a clinical perspective.","authors":"Ulrich Brinkmann, Roland E Kontermann","doi":"10.1080/19420862.2026.2613548","DOIUrl":"10.1080/19420862.2026.2613548","url":null,"abstract":"<p><p>Over the past two decades, bi- and multispecific antibodies have emerged as a rapidly advancing class of therapeutic biologics, transforming oncology and immunotherapy. By simultaneously binding two or more distinct antigens or epitopes, these molecules achieve mechanisms of action beyond those of conventional monoclonal antibodies, including immune cell redirection, dual pathway modulation, and enhanced tissue selectivity. Bispecific and multispecific antibodies exhibit considerable structural diversity, encompassing a wide range of molecular architectures covering a steady growing 'zoo' of formats. The therapeutic success and diversity of molecules and formats is reflected in the 2021 revision of the international nonproprietary name system, which introduced the suffix - mig to denote multispecific immunoglobulins. In this review, we provide an overview of multispecific antibodies in clinical development, focusing on format, molecular design, and clinical status. In total, data for 501 multispecific antibodies were compiled and analyzed, identifying 112 different formats. Overall, this analysis highlights the rapid growth, enormous format diversity, and translational potential of multispecific antibodies. It underscores their emerging role as versatile therapeutics not only in oncology, but also in non-cancer indications, reflecting a field that continues to evolve rapidly in response to both scientific innovation and clinical needs.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2613548"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12818813/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145989060","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction.","authors":"","doi":"10.1080/19420862.2026.2728528","DOIUrl":"10.1080/19420862.2026.2728528","url":null,"abstract":"","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2728528"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13577288/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148956222","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Overcoming claudin family homology: discovery of ARC101, a highly potent CLDN6-specific T-cell engager with a novel CD3 binder for ovarian adenocarcinoma.","authors":"Danlin Yang, Neha Kamran, Gururaj Shivange, Kavita Kumari, Prabhu Srinivas Yavvari, Viduth K Chaugule, Vishal Mahajan, Bridget Larkin, Scott R Brodeur, Joe Erhardt, Sanjaya Singh","doi":"10.1080/19420862.2026.2637299","DOIUrl":"10.1080/19420862.2026.2637299","url":null,"abstract":"<p><p>Claudin-6 (CLDN6) is an oncofetal tight junction protein with minimal to no expression in healthy adult tissues but aberrant upregulation in ovarian malignancies, making it an attractive tumor-selective antigen for T cell-based immunotherapy. The development of CLDN6-targeting antibodies, however, has been challenged by its high homology to CLDN9, which is expressed in normal tissues and differs by only three amino acids within the extracellular domains. Here, we describe the discovery and preclinical development of ARC101, a bispecific CLDN6×CD3 antibody featuring a naturally derived, highly potent CLDN6 binder with no cross-reactivity to CLDN9 or other human membrane proteins. The stringent specificity of ARC101 eliminates off-target binding and distinguishes it from other CLDN6-targeting antibodies in development. The effector arm of ARC101 incorporates a novel conformational CD3 binder, enabling potent T cell-mediated cytotoxicity against CLDN6-expressing tumor cells <i>in vitro</i> and <i>in vivo</i>. ARC101 also demonstrated a favorable pharmacokinetic profile in cynomolgus monkeys, low immunostimulatory responses in <i>ex vivo</i> human donor assays, and robust biophysical properties compatible with standard antibody manufacturing. Collectively, these findings support the clinical advancement of ARC101 as a differentiated, CLDN6-specific bispecific immunotherapy with exceptional tumor selectivity and optimized T cell activity for the treatment of solid tumors.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2637299"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12977278/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147390402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering of acidic pH-responsive anti-CD3 binding antibodies.","authors":"Grégory La Sala, Katharina B Kroell, Mudita Pincha, Christian Gassner, Lorenzo Deho, Ekkehard Moessner, Xavier Gueripel, Nicole Borin, Moritz Classen, Jörg Benz, Alexander Bujotzek, Christian Klein, Guy Georges, Adrian Hugenmatter, Klaus R Liedl, Anna Vangone","doi":"10.1080/19420862.2026.2658902","DOIUrl":"10.1080/19420862.2026.2658902","url":null,"abstract":"<p><p>The development of anti-CD3 antibody-based T cell engager therapeutics has improved the treatment of various malignancies, yet the challenge of achieving tumor-specific targeting while minimizing on-target off-tumor effects in normal tissues remains a substantial hurdle. One promising strategy to address this issue involves engineering antibodies with conditional pH-dependent binding affinities, capitalizing on the acidic microenvironment characteristics of tumors (pH ~ 6.5-6.8) compared to the neutral pH of healthy tissues (pH ~ 7.4). In this study, we focus on the pH-engineering of antibody binders against the human CD3 antigen, a critical component of T cell activation, to achieve preferential binding at acidic pH. Using molecular dynamics (MD) simulations on the reported CD3ɛ antibody binder 40G5c, we shed light on possible molecular mechanisms of the pH-responsiveness of key mutations and their impact on the overall binder structure at physiological or acidic pH. Our study highlights how MD has emerged as a powerful tool to guide and explain intrinsic pH-dependent molecular mechanisms in antibody engineering. Lastly, we report that our engineered CD3 binders preferentially bind and activate T cells under acidic pH conditions and display favorable affinity and pH-window profiles.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2658902"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13128029/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147775804","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel insulin-like growth factor II-based masking domain for conditional activation of therapeutic antibodies.","authors":"Lihua Shi, Minseon Cho, Motohiko Nishida, Mark L Chiu, Di Zhang","doi":"10.1080/19420862.2026.2668187","DOIUrl":"10.1080/19420862.2026.2668187","url":null,"abstract":"<p><p>Systemic target engagement constrains the therapeutic index of many antibody drugs, particularly in inflammatory disease and immuno-oncology. Protease-cleavable masking can localize activity to disease sites, but existing approaches often tradeoff masking strength, reversibility, and developability. Here, we evaluated the insulin-like growth factor II (IGF-II) as a compact, structurally defined steric masking domain for protease-dependent conditional activation of therapeutic antibodies. IGF-II masking domains, including a receptor-silent variant (IGF-II-s), were engineered as N-terminal fusions to therapeutic antibodies via protease-cleavable linkers and characterized using antigen-binding and cell-based functional assays. IGF-II masking attenuated anti-TNFα binding and neutralization potency in a protease-dependent manner and activity was largely restored following cleavage. <i>In vivo</i>, an MMP2/9-cleavable masked anti-TNFα antibody retained efficacy comparable to adalimumab and infliximab in a collagen antibody-induced arthritis model, yet, unlike the reference antibodies, did not measurably reduce survival in a <i>Listeria monocytogenes</i> infection challenge model at the dose tested. IGF-II-s showed no detectable IGF receptor binding and provided stronger masking than native IGF-II. This generalized approach was demonstrated across multiple antibodies, enabling efficient protease-dependent conditional activation of trastuzumab (anti-HER2), an anti-IL-1β antibody, and bevacizumab (anti-VEGF). Together, these results established an engineered IGF-II-based masking domain as a versatile, IGF-II receptor-silent platform enabling protease-dependent conditional activation of therapeutic antibodies, with potential to improve safety and expand therapeutic windows.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2668187"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13166275/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RASP: rapid antibody functional screening by pentavalent phage display.","authors":"Manpreet Kaur, Abhishek Dubey, Kartik Chandran","doi":"10.1080/19420862.2026.2627708","DOIUrl":"10.1080/19420862.2026.2627708","url":null,"abstract":"<p><p>The heavy-chain antibody variable domain (VHH) is the smallest antigen-binding domain of such antibodies, which are derived from camelids. In the past three decades, VHHs, which are also called single-domain antibodies, have been extensively used to target pathogens and/or toxins. Conventional screening methods, such as phage display, rely only on antibody-antigen binding as the sole criterion for selection. Despite being robust and high-throughput, such methods often require additional downstream experiments to identify VHH that neutralize their target. Here, we describe an innovative, high-throughput functional screening method, Rapid Antibody functional Screening by Pentavalent phage display (RASP), that incorporates purified antibody-displaying phages for virus neutralization assays, and thus can be used to directly identify neutralizing VHHs. As a proof-of-concept, we first displayed previously identified neutralizing VHHs specific for the spike proteins of SARS-CoV-2 and respiratory syncytial virus on phages and demonstrated a dose-dependent blockade of viral infection. We further improved our method by utilizing the pentavalent display feature of hyperphages. We showed that hyperphage-derived VHH phages were superior to helper phage-derived VHH phages in assaying viral neutralization potential. Thereafter, we applied RASP to identify multiple candidates by screening a semi-synthetic VHH library against recombinant vesicular stomatitis viruses pseudotyped with spike glycoproteins from SARS-CoV-2, Junin virus, and Ebola virus, featuring as case studies in antiviral antibody discovery. Further, we benchmarked RASP against established phage ELISA and next-generation sequencing methods. Overall, we successfully used RASP in the context of the discovery of antiviral VHHs, highlighting its broader applicability as a platform that can be used either in isolation or in conjunction with other functional screening methods to accelerate the discovery of antiviral VHHs.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2627708"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12928633/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146143021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}