Antibodies最新文献

{"title":"Discovery of Anti-SARS-CoV-2 XBB.1.5 and JN.1 Variant-Specific Monoclonal Single-Domain Antibodies from a Synthetic Library.","authors":"Isamu Tsuji, Kumiko Okada, Benjamin Kroppen, Tetsufumi Katta, Kaori Yamamura, Takeshi Nishihama, Ayako Miura, Hansjörg Götzke, Eric Crampon, Andrea Bertolotti-Ciarlet","doi":"10.3390/antib15020018","DOIUrl":"10.3390/antib15020018","url":null,"abstract":"<p><p><b>Background/Objectives:</b> The SARS-CoV-2 virus frequently undergoes mutations to evade the human immune system. Vaccines for new strains are developed each season, and an identification test confirming the specific strain is essential for vaccine quality control, as stated by the U.S. Food and Drug Administration. However, a shorter timeline of antibody discovery was required to adjust vaccine development schedules. Therefore, anti-SARS-CoV-2 strain-specific, single-domain antibodies (sdAbs) for SARS-CoV-2 vaccines were discovered using alpaca synthetic libraries without animal immunization. <b>Methods:</b> A synthetic sdAb library was developed based on conserved alpaca sdAb frameworks, with a degree of freedom in the three complementarity-determining regions. Specific and high-affinity sdAb clones were selected from the library by one ribosomal display round, followed by two phage display selections using a biotinylated strain-specific SARS-CoV-2 receptor-binding domain (RBD) of the spike protein as bait and non-biotinylated RBD variants to block. The sdAbs clones were applied to the identification test using Western blotting. The binding epitopes were determined by hydrogen-deuterium exchange mass spectrometry. <b>Results:</b> Five clones of XBB.1.5 and two clones of JN.1-specific sdAbs were discovered. Anti-JN.1 sdAb clone 1B9 detected JN.1 vaccine products but no other previously produced vaccine strains, Wuhan, BA.5 and XBB.1.5, by WB for vaccine identification test. Four binding epitopes for anti-JN.1 sdAb clone 1B9 were identified, including the L455S mutation, a critical amino acid to evade neutralizing antibodies for the JN.1 strain. <b>Conclusions:</b> Anti-XBB.1.5 and JN.1-specific sdAbs were discovered from a synthetic single-domain antibody library within 8-9 weeks, and these sdAbs were applied to vaccine identification testing.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 2","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13010742/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147503124","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}

{"title":"Microsecond Dynamics of Fc-CD16a Recognition: Impact of Mutations, Core Fucosylation, and Fc Asymmetry.","authors":"Sébastien Estaran, Bernard Hehlen, Alain Chavanieu","doi":"10.3390/antib15010017","DOIUrl":"10.3390/antib15010017","url":null,"abstract":"<p><strong>Background/objectives: </strong>Antibody-dependent cellular cytotoxicity relies on the interaction between the Fc region of immunoglobulin G1 (IgG1) and the CD16a receptor. While removal of core fucosylation on Fc and introduction of the DFTE mutation set (S239D, H268F, S324T, I332E) are known to enhance CD16a binding, the detailed contributions of these engineered sites in solution remain incompletely defined.</p><p><strong>Methods: </strong>Here, we employed 1 µs molecular dynamics simulations to map, at atomic resolution, the interaction networks stabilizing pre-formed Fc-CD16a complexes, including afucosylated Fc-wild-type, DFTE-engineered, Fc-fucosylated, and asymmetrically engineered Fc variants.</p><p><strong>Results: </strong>Our results show that only S239D, present on both Fc chains, and H268F on chain A consistently contribute to stabilizing the CD16a interface, while I332E does not form persistent interactions. Glycan-protein contacts are primarily intrachain, with transient interchain glycan-glycan interactions not contributing significantly to complex stability. Fucosylation on Fc significantly reduces binding stability by disrupting peripheral interactions and critical glycan-mediated contacts. Notably, the asymmetric Fc variant, in which the two heavy chains carry distinct sets of substitutions, retains high-affinity binding despite lacking S239D and carrying core fucose, through a novel hydrophobic cluster and reinforced peripheral electrostatic interactions.</p><p><strong>Conclusions: </strong>Altogether, these findings provide a quantitative framework for how targeted mutations and fucose modifications remodel Fc-CD16a interactions, offering insights for the rational design of next-generation therapeutic antibodies.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12937375/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147288925","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}

{"title":"Analytical Performance of Nanobody-Based Immunoassay and Immunosensing Platforms for Bacteria and Toxin Detection: A Systematic Review.","authors":"Aya Jalil, Nadia Touil, Omar Nyabi, Elmostafa El Fahime, Sara Benlhachemi, Jean-Luc Gala, Khalid Ennibi, Karim Bakkouri, Abdelaziz Benjouad, Lamiae Belayachi","doi":"10.3390/antib15010015","DOIUrl":"10.3390/antib15010015","url":null,"abstract":"<p><p><b>Background:</b> bacterial pathogens and their toxins present analytical challenges for rapid and specific detection, contributing to over 600 million cases of illness annually and worsening antimicrobial resistance (AMR). Conventional detection methods are useful but limited. Single-domain antibodies (sdAbs) offer alternative recognition elements with unique biochemical and engineering benefits, enabling the development of nanobody-based immunoassays and biosensing platforms that provide fast, highly selective, and reliable detection of bacterial pathogens and toxins in both food and clinical environments. <b>Objectives:</b> this systematic review assesses the analytical and functional performance of nanobody-based immunoassays and sensing formats for detecting bacteria and toxins across food and clinical samples. <b>Methods:</b> following PRISMA guidelines, major scientific databases were used to gather research, resulting in 32 eligible studies published between 2011 and 2025. <b>Results:</b> data collected included assay platforms, target bacteria and toxins, limit of detection, sensitivity, specificity, matrix recovery, and practicality. Risk of bias was evaluated using an adapted QUADAS-2 framework. The review shows that nanobody-based immunoassays have achieved high performance, thermostability, compatibility with genetic engineering, and versatile assay design. When combined with advanced transduction and signal amplification strategies, these systems contribute to the development of highly sensitive and user-friendly bioanalytical platforms for detecting bacteria and toxins. <b>Conclusions:</b> however, most studies relied on spiked samples and lacked large-scale validation, emphasizing the need for standardized benchmarking and real-world testing.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12937825/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147288790","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}

{"title":"Generation of Schlafen 8-Specific Antibodies.","authors":"Juan Carlos Silva-Espinoza, Mauricio I Rodriguez Rodriguez, Claire Eunise Perucho, Brian A Terrazas, Carlos Valenzuela, Stephany Palos Vargas, Andrea Carlin, Diana L Prospero, Giulio Francia, Manuel Llano","doi":"10.3390/antib15010016","DOIUrl":"10.3390/antib15010016","url":null,"abstract":"<p><p><b>Background/Objectives</b>: Schlafen (SLFN) 8 and SLFN9 are mouse members of the Schlafen protein family, believed to have arisen through a gene duplication event. The physiological roles of these proteins remain poorly defined, in part due to the absence of reliable, commercially available antibodies for their detection. <b>Methods</b>: To develop specific antibodies, we performed an amino acid sequence alignment of these proteins and identified a thirteen amino acids long peptide predicted by AlphaFold modeling and hydropathicity analysis to be surface-exposed in both SLFN proteins. The SLFN8 peptide was conjugated to KLH and used to immunize mice, employing Poly(I:C) as an adjuvant. <b>Results</b>: We verified the anti-SLFN8 antibody specificity in mouse tissues, engineered human cells, and recombinant proteins by different immunodetection techniques, including Western blotting, immunoprecipitation, immunohistochemistry, and ELISA. Furthermore, splenocytes from immunized mice were used to generate hybridomas that secreted IgG antibodies with SLFN8-peptide specificity, as assumed by ELISA. <b>Conclusions</b>: Our results demonstrate that the identified peptide is highly immunogenic and capable of eliciting antibodies that distinguish between these two exceedingly similar proteins in a broad group of immunodetection techniques.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12938254/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147288769","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}

{"title":"Transplacental Antibody Transfer: Mechanisms, Pregnancy-Related Disruptions, and Emerging Experimental Models.","authors":"Qiqi Li, Zhengyuan Huang, Zainab Saeed, Orene Greer, James A Harker, Nishel M Shah","doi":"10.3390/antib15010014","DOIUrl":"10.3390/antib15010014","url":null,"abstract":"<p><p>The transplacental transfer of maternal immunoglobulin G from the mother to the foetus is central for providing immunity in early life, resulting in full-term newborns having IgG repertoires and levels similar to those of their mothers. The neonatal Fc receptor is recognised as the primary transporter of IgGs across the placental epithelium. Understanding the mechanisms of transplacental antibody transfer and factors that affect them is essential in optimising maternal vaccination strategies, ultimately protecting infants from various environmental pathogens. This review first outlines the biological mechanisms governing transplacental IgG transfer, followed by a discussion of how this process may be disrupted by physiological and pathological conditions during pregnancy, including preterm birth, hypergammaglobulinemia, maternal pathogenic IgG, maternal infections, hyperglycaemia, and exposure to biological therapies. We also summarise currently available models used to study transplacental IgG transfer, highlighting existing knowledge gaps and future directions for research in this field.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12922050/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146257111","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}

{"title":"Antibody Avidity Profiles as Diagnostic Biomarkers in Differentiating Acute and Chronic <i>Anisakis simplex</i>-Related Allergic Diseases.","authors":"Juan González-Fernández, Laura Ullate, Marta Rodero, Alvaro Daschner, Carmen Cuéllar","doi":"10.3390/antib15010013","DOIUrl":"10.3390/antib15010013","url":null,"abstract":"<p><strong>Background/objectives: </strong>Allergic features of anisakiasis, caused by ingestion of third-stage larvae of <i>Anisakis simplex</i> via raw or undercooked fish, manifest clinically as acute gastroallergic anisakiasis (GAA) or chronic urticaria with <i>Anisakis</i> sensitization (CU+). Differentiating these clinical phenotypes remains challenging. This study aimed to evaluate the maturation and avidity of specific antibodies (IgE, IgG4, IgG, and IgA) as biomarkers for discriminating between acute and chronic forms of anisakiasis.</p><p><strong>Methods: </strong>A prospective cohort of 65 patients from Madrid, Spain, was classified into three groups: GAA (n = 22), CU+ (n = 22), and chronic urticaria without sensitization (CU-, n = 21). Serum samples were analyzed for antigen-specific immunoglobulins using ELISA and Western blot. Avidity indices (AIs) were quantified through urea dissociation assays. Statistical comparisons and correlation analyses were performed to associate antibody avidity with clinical phenotype and demographic variables.</p><p><strong>Results: </strong>GAA patients exhibited significantly lower IgE avidity indices compared to CU+ individuals (mean AI: 79.9% vs. 88.5%), indicating a less mature IgE response during acute infection. Conversely, IgG4 and IgG avidity were elevated in GAA relative to CU+, reflecting an active but transient immune response. IgA antibodies were detected in both groups, although avidity differences lacked discriminatory capacity. No sex- or age-related differences in antibody avidity were observed. Longitudinal follow-up of GAA patients demonstrated an increase in IgE avidity over time.</p><p><strong>Conclusions: </strong>Quantitative assessment of antibody avidity, particularly for IgE and IgG4, enhances understanding of <i>A. simplex</i> immunopathogenesis and serves as a valuable biomarker for distinguishing acute from chronic clinical presentations. These findings support the use of avidity indices in the diagnosis, staging, and clinical management of anisakiasis.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921752/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146257024","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}

{"title":"Comparative In Vitro Evaluation of Anti-HIV Immunotoxin, Antibody-Drug Conjugate, and Radioimmunoconjugate Targeted by the Same Antibody.","authors":"Anne-Sophie Kuhlmann, Tami Peters, Donald K Hamlin, Yawen Li, Xinyi Wang, Megan Stackhouse, Frances M Cole, Jasmin Martinez-Reyes, Brenda M Sandmaier, Hans-Peter Kiem, D Scott Wilbur, Robert D Harrington, Seth H Pincus","doi":"10.3390/antib15010012","DOIUrl":"10.3390/antib15010012","url":null,"abstract":"<p><strong>Background: </strong>We are developing cytotoxic immunoconjugates (CICs) to eliminate HIV-infected cells. We investigated the efficacy and kinetics of killing by different forms of CICs targeted by the same monoclonal antibody (mAb), an immunotoxin (IT), antibody-drug conjugate (ADC), and radioimmunoconjugate (RIC).</p><p><strong>Methods: </strong>We compared in vitro effects of CICs made by conjugating anti-gp41 mAb 7B2 to deglycosylated ricin A chain (7B2-dgA), the anthracycline derivative PNU-159682 (7B2-PNU), or the α-emitting isotope actinium-225 (7B2-²²⁵Ac). Kinetic analyses of cell growth were performed measuring electrical impedance every 15 min over a 7-day period using cells stably expressing the HIV envelope and Env-negative parent cells.</p><p><strong>Results: </strong>7B2-dgA and 7B2-²²⁵Ac were more potent and acted more rapidly to kill cells than 7B2-PNU. Both the 7B2-PNU and 7B2-²²⁵Ac induced bystander-cell killing, whereas the IT did not and consequently allowed the outgrowth of Env-negative cells. Low dose or brief exposure to 7B2-PNU resulted in an increased rate of cell growth.</p><p><strong>Conclusions: </strong>An IT, ADC, and RIC showed substantial differences in the degree of specific toxicity, kinetics, and mechanisms of killing. The results of this side-by-side comparison have implications for the development of CICs to treat HIV, as well as other conditions.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921964/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146257051","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}

{"title":"Antibody Screening and Binding Prediction Analysis Targeting Stx2.","authors":"Jilei Wu, Chenghua Liu, Fenghao Peng, Zeyuan Yu, Chunxia Qiao, Guang Yang, Heng Luo, Keyi Sun, Ziyao Ning, Jing Wang, Yan Wen, Jijun Yu","doi":"10.3390/antib15010011","DOIUrl":"10.3390/antib15010011","url":null,"abstract":"<p><p><b>Background</b>: Shiga toxin (Stx), produced by enterohemorrhagic <i>Escherichia coli</i> (EHEC), is a highly potent exotoxin responsible for severe complications such as hemolytic uremic syndrome (HUS). Among its isoforms, Stx2 exhibits stronger cytotoxicity and poses greater clinical risk, yet no effective therapy currently exists. <b>Methods</b>: In this study, two human monoclonal antibodies, YG12-1 and YG12-2, were identified from a phage display library and systematically characterized using an integrated modeling-validation workflow. <b>Results</b>: Structural modeling with ImmuneBuilder and Rosetta revealed that YG12-2 possessed a longer CDRH3 topology, more short-range hydrogen bonds, and stronger electrostatic complementarity, corresponding to lower binding energy and higher apparent affinity in ELISA and SPR. Although YG12-2 had a better affinity, YG12-1 shows better protective activity in a murine model of acute peritoneal infection. This paradox highlights a non-linear relationship between structural affinity and biological efficacy, emphasizing the importance of functional epitope accessibility and pharmacokinetic behavior in determining neutralization outcomes. <b>Conlusions</b>: Overall, these results indicated that targeting Stx2 with YG12-1 and YG12-2 could serve as a promising protective strategy against <i>E. coli</i> O157:H7 infection.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921925/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146257077","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}

{"title":"Antibodies as Tools for Characterization, Isolation and Production Enhancement of Anti-Cancer Drugs and Steroidal Hormones from Ginsenoside and Solasodine Glycoside: A Review.","authors":"Yukihiro Shoyama","doi":"10.3390/antib15010010","DOIUrl":"10.3390/antib15010010","url":null,"abstract":"<p><p>There are a vast number of monoclonal antibodies (MAbs) against biological components; however, the number for natural products is less than 50. MAbs against ginsenosides, i.e., dammarane triterpene glycosides contained in ginseng, were prepared to develop an Eastern blotting method that can estimate the number of bound sugars and pharmacological activity. Meanwhile, as a method for producing ginsenoside Rg3, which is used as an anti-cancer drug, an affinity column for ginsenoside Rb1 was prepared to isolate the raw material ginsenoside Rb1 in a single step, and a method for obtaining ginsenoside Rg3 through fermentation was proposed. A unique MAb capable of detecting all solasodine glycosides contained in <i>Solanum</i> plants was created to prepare an affinity column capable of isolating solasodine glycosides from <i>S. khasianum</i> fruit in a single step. The single-chain variable fragment gene was induced from the MAb against solasodine glycoside and introduced into the hairy root system of <i>S. khasianum</i>, thereby increasing the solasodine glycoside content more than twofold. As a result, we recognized that this method can be used to breed plants with higher concentrations of plant secondary metabolites like solasodine glycosides. The above results collectively demonstrate that solasodine glycoside can be isolated from <i>S. khasianum</i> in high yields and that this compound enables the production of steroids in high yields through a one-step chemical reaction.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921994/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146257053","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}

{"title":"Robustness Evaluation of a Legacy N-Glycan Profiling Method for a Therapeutic Antibody Under ICH Q14 Lifecycle Principles.","authors":"Ming-Ching Hsieh, Chao Richard Li, Margaret A Velardo, Jingming Zhang, Babita S Parekh","doi":"10.3390/antib15010009","DOIUrl":"10.3390/antib15010009","url":null,"abstract":"<p><strong>Background: </strong>This study assesses the robustness of a legacy N-glycan profiling method for the therapeutic antibody MAB1 with different Peptide-N-glycosidase F (PNGase F) enzyme sources, solid phase extraction (SPE) cartridges, and reagent stability, aligning with ICH Q14 lifecycle management principles. Glycosylation profiling is critical for therapeutic antibodies as it influences both function and pharmacokinetics.</p><p><strong>Method: </strong>The legacy N-glycan profiling method, 2-aminobenzoic acid (2-AA) labeling combined with normal-phase HPLC, was re-evaluated to confirm consistent analytical performance in the context of evolving regulatory expectations. The evaluation focused on three key factors: PNGase F enzyme sources, solid-phase extraction (SPE) cartridges, and reagent stability.</p><p><strong>Results: </strong>Commercial PNGase F enzymes showed various performances, with some sources yielding significant differences. Several SPE cartridges were also tested, with certain formats displaying poor recovery and high variability, particularly for sialylated glycans. In addition, reagent stability studies revealed rapid degradation of the labeling reagent within a few days.</p><p><strong>Conclusions: </strong>These results underscore the importance of risk control, continual improvement, and lifecycle management to ensure reliable glycosylation analysis and the sustained robustness of legacy methods.</p>","PeriodicalId":8188,"journal":{"name":"Antibodies","volume":"15 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12821501/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146008187","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}