mAbs最新文献

{"title":"NanoSaurus: addressing the urgent need for open access in single-domain antibody research.","authors":"P Di Filippo, E Pardon, R J Owens, F I Schmidt, C Durinx, U Rothbauer, H Ploegh, M A Seeger, S Muyldermans, W Vranken, J Steyaert","doi":"10.1080/19420862.2026.2733370","DOIUrl":"10.1080/19420862.2026.2733370","url":null,"abstract":"","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2733370"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13625707/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149074738","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":"HDX-MS-Guided computational modeling for rapid and accurate antibody-antigen interface mapping.","authors":"Will McMahon, Hanzhi Zhang, Xiuling Li, Vaheh Oganesyan, Sterling Payne, Nurjahan Mehzabeen, Nydia van Dyk, Wenhai Liu, Wesley Howard, Pengyi Zhao, Yeying Zhang, Mulin Fang, Xiaoyu Chen, Yariv Mazor, Yu Shi","doi":"10.1080/19420862.2026.2722455","DOIUrl":"10.1080/19420862.2026.2722455","url":null,"abstract":"<p><p>Accurate mapping of paratopes and epitopes at antibody-antigen (Ab-Ag) interfaces is essential for the rational design of therapeutic antibodies in early drug development. Traditional methods for determining high-resolution Ab-Ag complex structures, such as X-ray crystallography and cryo-electron microscopy, are labor-intensive and require substantial time and resources. Although recent computational approaches have increased throughput, prediction accuracy and reliability remain limited, particularly without experimental structural constraints. Here, we present a hydrogen-deuterium exchange mass spectrometry (HDX-MS)-guided structural modeling workflow for efficient and accurate epitope and paratope mapping across three prospective case studies in therapeutic protein discovery and engineering. Across these systems, peptide-level HDX-MS data were sufficient to guide docking and structural modeling, whereas targeted electron transfer dissociation (ETD) HDX-MS was applied in one system as a higher-resolution, orthogonal approach to further localize critical binding residues. The workflow was applied to diverse Ab-Ag architectures spanning Fab and VHH binders and homodimeric and heterodimeric antigen ectodomains, and HDX-MS data were integrated with HADDOCK, AlphaFold2, and Protenix to generate Ab-Ag complex models. Comparison with subsequently obtained in-house X-ray crystal structures, used here as orthogonal high-resolution reference structures, showed strong agreement in overall binding orientation and interface placement. These results demonstrate the robustness and practical utility of this integrative approach for structure-informed antibody engineering in therapeutic protein discovery.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2722455"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13577250/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148945206","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":"Predicting non-specific binding of VHHs using machine learning models with cluster-aware validation.","authors":"Valentin Stanev, Federico Devalle, Mehdi Boroumand, Maryam Pouryahya, Isabelle Sermadiras, Jenna Caldwell, Kuan-Lin Chen, Jay Hyun Jo, Rohan Jain, Bismark Amofah, Tony Pham, Mark Hutchinson, Sharfa Farzandh, Jen DiChiara, Chacko Chakiath, Tom Diethe, Andrew Dippel, Gilad Kaplan, Rebecca Croasdale-Wood","doi":"10.1080/19420862.2026.2732787","DOIUrl":"10.1080/19420862.2026.2732787","url":null,"abstract":"<p><p>Propensity for nonspecific binding-also known as polyreactivity-is a serious developability risk factor for biotherapeutic candidates. To minimize this risk, drug companies are increasingly relying on <i>in silico</i> tools utilizing machine learning methods, but developing these tools is challenging. For example, the available data often contains many closely related sequences originating from drug pipeline projects, which can introduce significant biases in the <i>in silico</i> models training and benchmarking, leading to poor generalizability on new data. We present here a workflow designed to diagnose and mitigate some of the problems associated with using pipeline data. The workflow is based on a custom cross-validation procedure that can evaluate model performance on unseen data in different contexts. As a demonstration of the workflow, we use it to train a model to predict variable heavy-chain only fragment antibodies (VHH) binding to baculovirus particles (BVP)-a widely used assay for nonspecific binding. Using descriptors based on computed protein structures, the workflow identifies several risk factors that correlate with higher polyreactivity levels.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2732787"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13596884/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148982770","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":"Beyond sequence similarity: ML-powered identification of pHLA off-targets for TCR-mimic antibodies using high throughput binding kinetics.","authors":"Alexander Sinclair, Stefan Krämer, Christoph Reinhart, Jennifer Stehle, Simon Schuster, Tobias Herz, Hoor Al Hasani, Pranav Hamde, Oliver Selinger, Joerg Birkenfeld","doi":"10.1080/19420862.2025.2601360","DOIUrl":"10.1080/19420862.2025.2601360","url":null,"abstract":"<p><p>T-cell receptor mimic (TCRm) antibodies are an emerging class of tumor-targeting agents used in advanced immunotherapies such as bispecific T-cell engagers and CAR-T cells. Unlike conventional antibodies, TCRms are designed to recognize peptide - human leukocyte antigen (pHLA) complexes that present intracellular tumor-derived peptides on the cell surface. Due to the typically low surface abundance and high sequence similarity of pHLAs, TCRms require high affinity and exceptional specificity to avoid off-target toxicity. Conventional methods for off-target identification such as sequence similarity searches, motif-based screening, and structural modeling focus on the peptide and are limited in detecting cross-reactive peptides with little or no sequence homology to the target. To address this gap, we developed EpiPredict, a TCRm-specific machine learning framework trained on high-throughput kinetic off-target screening data. EpiPredict learns an antibody-specific mapping from peptide sequence to binding strength, enabling prediction of interactions with unmeasured pHLA sequences, including sequence-dissimilar peptides. We applied EpiPredict to two distinct TCRms targeting the cancer-testis antigen MAGE-A4. The model successfully predicted multiple off-targets with minimal sequence similarity to the intended epitope, many of which were experimentally validated via T2 cell binding assays. These findings establish EpiPredict as a valuable tool for lead optimization of TCRms, enabling the identification of antibody-specific off-targets beyond the scope of traditional peptide-centric methods and supporting the preclinical de-risking of TCRm-based therapies.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2601360"},"PeriodicalIF":7.3,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12710896/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145743226","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":"Fine-tuning affinity and spacer design enhances T cell potency in DLL3 and BCMA CAR T cells.","authors":"Nicholas Mazzanti, Ninkka Tamot, Andrea Francese, Jinquan Luo, M Jack Borrok, Julie Rossillo, Joseph Plummer, Gauri Anand Patwardhan, Chi Shing Sum, Michael Ports, Kara L Spiller, Madhusudhanan Sukumar","doi":"10.1080/19420862.2025.2602989","DOIUrl":"10.1080/19420862.2025.2602989","url":null,"abstract":"<p><p>Chimeric antigen receptor (CAR)-modified T cells have garnered substantial attention due to their clinical success, culminating in six Food and Drug Administration-approved therapies for hematological malignancies. Notably, CD19-specific CAR T cell therapies have achieved remarkable clinical efficacy in treating B-cell malignancies, but these profound and durable responses are not observed in CAR T therapies targeting other indications, particularly solid tumors. Key design elements of CAR constructs - namely, antigen binding affinity and spacer length - play critical roles in determining T cell effector function and overall therapeutic effectiveness. Refining CAR designs may enhance T cell functionality, extend clinical application, and potentially apply CAR T cell therapies across a wider array of malignancies. In this study, affinity variant and spacer variant CARs targeting BCMA and DLL3 tumor antigens were evaluated using <i>in vitro</i> measurements of antigen-binding properties and effector function. Each panel of CARs spanned 2-3 logs of antigen binding affinity (BCMA: 181 pM KD to 74 nM KD, DLL3: 417 pM to 407 nM). Additionally, CAR T cells were challenged with tumor spheroids composed of BCMA⁺ H929 and DLL3⁺ SHP77 tumor cells. We show that for both tumor models, higher affinity CARs (KD stronger than approximately 100 nM) paired with an intermediate length spacer (IgG1 Fc, CH2-CH3, 230AA) elicited the strongest levels of tumor killing, CAR⁺ T cell expansion, and proinflammatory cytokine production. These CARs displayed the strongest cellular affinity when measured in a conjugation assay, suggesting a relationship between cellular affinity and T cell functional performance. This study highlights the critical role of CAR design in enhancing T cell functionality, demonstrating that high-affinity CARs combined with intermediate-length spacers yield superior performance in targeting BCMA and DLL3 antigens. This study provides a framework for rational CAR design, informing strategies to broaden the clinical utility of CAR T-cell therapies beyond hematologic cancers.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2602989"},"PeriodicalIF":7.3,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12710886/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145743209","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":"Systematic characterization of lysine glucuronidation in a bispecific antibody.","authors":"Michael R Reyda, Qinqin Ji, Maggie Huang, Izabela Sokolowska, Qingrong Yan, Joseph Mulholland, Jingjie Mo, Ping Hu","doi":"10.1080/19420862.2025.2612471","DOIUrl":"10.1080/19420862.2025.2612471","url":null,"abstract":"<p><p>This study presents a systematic characterization of lysine glucuronidation that was revealed during the charge variant characterization of a bispecific antibody (bsAb). Site-specific quantitation by Glu-C/Asp-N peptide mapping suggested that glucuronidation occurred randomly across surface lysine residues. To understand the impact of glucuronidation on the structure and function of the bsAb, stressed samples with up to 84% total glucuronidation were generated and analyzed by a comprehensive panel of analytical methods. The results suggested that glucuronidation caused an acidic isoelectric point (pI) shift in the charge profile. However, it does not affect the higher-order structure or bioactivities of the bsAb, including antibody-dependent cell-mediated cytotoxicity, antigen binding, or Fc receptor interaction. To support routine process monitoring, a fit-for-purpose subunit mass method was developed and qualified for quantitation of glucuronidation, offering a higher-throughput alternative to peptide mapping for assessing process consistency and product comparability.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2612471"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12795262/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145933908","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":"Structures of clinical antibodies bound to IL-33 uncover two distinct epitopes underlying differential efficacy.","authors":"Jing Chen, Yue Wang, Xinquan Wang","doi":"10.1080/19420862.2026.2639673","DOIUrl":"10.1080/19420862.2026.2639673","url":null,"abstract":"<p><p>Interleukin-33 (IL-33), an alarmin cytokine of the IL-1 family, drives type 2 inflammation through signaling via the ST2 and IL-1RAcP receptors, making it a critical therapeutic target for inflammatory diseases such as asthma and chronic obstructive pulmonary disease. Current therapeutic strategies have primarily focused on antibodies that target IL-33 or ST2 to disrupt their specific interaction. However, the structural mechanisms underlying antibody-mediated neutralization of IL-33 remain poorly understood. Here, we report the structures of three antibodies in clinical trial - etokimab, itepekimab, and tozorakimab - complexed with IL-33, determined by X-ray crystallography and cryo-electron microscopy. Structural analysis reveals two distinct neutralizing epitopes on IL-33, termed Epitope 1 at IL-33/ST2 binding Site 1 and Epitope 2 at IL-33/ST2 binding Site 2. Tozorakimab, which targets Epitope 1, completely blocks ST2 engagement by sterically occluding the ST2 D1-D2 domain-binding interface. In contrast, etokimab and itepekimab, which recognize Epitope 2, interfere with IL-33 recognition of the ST2 D3 domain and thereby only partially inhibit ST2 binding. These structural and biochemical findings provide a molecular explanation for the differential efficacy of the three antibodies in inhibiting IL-33 signaling in cellular assays. Collectively, our results provide valuable insights into the molecular determinants of efficacy for existing IL-33 therapeutics and offer a structural framework for the rational design of next-generation IL-33 targeted inhibitors.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2639673"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12959182/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147326608","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 cell line development vector strategy for improved expression of a trispecific T-cell engager in CHO.","authors":"Rajesh K Mistry, Chendi Nui, Giulia Lambiase, Daniel Ray, Lewis Kearsey, Noah Hitchcock, Luigi Grassi, Ramy Elgendy, James Fleming, Alexandra C Broughton, Peng Zhao, Chi-I Chiang, Pooja Shah, Matthew Cyr, Even Walseng, Yariv Mazor, Diane Hatton, Sarah Dunn","doi":"10.1080/19420862.2026.2632994","DOIUrl":"10.1080/19420862.2026.2632994","url":null,"abstract":"<p><p>Recent advances in trispecific antibody (trisAb) engineering offer great therapeutic potential, but achieving high product yield and quality in cell line development remains a challenge due to complex chain pairing requirements in production cell lines. In this study, three distinct expression vector configurations were evaluated for their ability to support robust, high-level expression of a structurally complex, synapse-gated trisAb T-cell engager (TriMab) in stable Chinese hamster ovary cells. Initial configurations using conventional dual heavy chain (HC) and triple light chain (LC) vectors resulted in poor pool performance characterized by delayed transfection recovery and low titers. By contrast, a redesigned strategy that reversed HC gene order and distributed LCs over separate vectors markedly improved transfection recovery along with product titers and reduced the formation of undesired product variants. Clonal cell lines established with this optimized strategy achieved titers exceeding 2 g/L with correct product quality profiles. Gene copy number and mRNA analyses confirmed that chain order and vector design strongly influenced mRNA levels and thus productivity. These results highlight the critical impact of vector configuration on manufacturability of complex TriMabs, providing a practical framework for the rational design of gene vectors to support next-generation trisAb production.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2632994"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12947573/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147307356","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":"Discovery and optimization of a pH-responsive ultra-long-acting VHH-based growth hormone mimetic.","authors":"Houcong Jin, Gao Sun, Hongxia Shen, Qian Hao, Lanlan Chen, Chong Che, Xiaoyu Gu, Tong Wang, Nizhi Wang, Fangfang Du, Qingjuan Ma, Peng Qi, Mengyao An, Xuemei Liao, Xiaozhen Wang, Siqin Wang, Lei Jin, Lishan Kang","doi":"10.1080/19420862.2026.2691350","DOIUrl":"10.1080/19420862.2026.2691350","url":null,"abstract":"<p><p>Weekly formulations of long-acting growth hormone (LAGH) effectively treat growth hormone (GH) deficiency in both children and adults while offering improved convenience compared to daily GH. To further enhance patient convenience and compliance, longer-acting GH preparations have become a key research focus. However, due to the inherent short half-life of protein-based drugs, no ultra-long-acting growth hormone or analog products (with dosing intervals longer than once weekly) are currently available worldwide. In this study, we developed a potentially ultra-long-acting growth hormone receptor (GHR) agonist pH.VHH03 with GH-mimetic activity by engineering hinge region and variable region of a VHH antibody. Through optimization of the hinge region, the <i>in vitro</i> cell proliferation activity of this molecule was significantly enhanced. As a result, it demonstrated LAGH-like effects in promoting body weight gain and stimulating IGF-1 secretion in rats. Furthermore, pH-dependent binding was introduced into the variable region. The optimized molecule exhibited a 227-fold difference in dissociation rates between neutral and acidic conditions. This modification prolonged its <i>in vivo</i> efficacy in rats from 6 days to over 15 days, far exceeding the 3-day duration observed with PEGylated GH. Subsequent <i>in vivo</i> experiments in rats confirmed that the final optimized molecule dose-dependently promoted body weight gain, insulin-like growth factor 1 (IGF-1) secretion, tibial growth, and significantly increased growth plate thickness. The <i>in vivo</i> profile fully replicated the physiological activities of GH. Moreover, at medium to high doses, it induced a notably flatter and sustained IGF-1 response compared to PEGylated GH, suggesting a potentially longer duration of pharmacological activities in humans.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2691350"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13313243/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148317946","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":"One-step cleavage and purification: a bifunctional IdeS-Protein A fusion protein for on-column generation of antibody fragments.","authors":"Tongsheng Zhang, Huayuan Liao, Chun-Bo Teng, Chao Wang","doi":"10.1080/19420862.2026.2684388","DOIUrl":"10.1080/19420862.2026.2684388","url":null,"abstract":"<p><p>Conventional production of antibody fragments (e.g. Fab/F(ab')₂, VHH/(VHH)₂) involves sequential enzymatic cleavage and multi-step purification to remove Fc domains and residual enzymes, a process that is laborious and inefficient for small-scale workflows. To address this, we engineered a bifunctional IdeS-Protein A (IPA) fusion protein, which combines the specific IgG-cleaving activity of IdeS with the Fc-binding capability of Protein A. In solution, this construct enabled simultaneous antibody cleavage and affinity-mediated removal of both the enzyme and Fc fragments in a single step. Furthermore, we immobilized the fusion protein on a chromatographic resin to create an integrated \"on-column cleavage-purification\" platform. This system allows antibody capture, in-situ cleavage, and direct collection of high-purity F(ab')₂ fragments from the flow-through, while Fc fragments, uncleaved antibodies, and the enzyme are retained on the column. This work establishes a streamlined, one-step method for generating antibody fragments, offering a robust and automatable platform for research and preclinical applications.</p>","PeriodicalId":18206,"journal":{"name":"mAbs","volume":"18 1","pages":"2684388"},"PeriodicalIF":7.9,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13245094/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148163747","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}