Protein Science最新文献

{"title":"Conformational determinants of glucagon stability and aggregation kinetics in aqueous and commercial formulations.","authors":"Angelo Santoro, Marco Macis, Michela Buonocore, Antonio Ricci, Anna Maria D'Ursi","doi":"10.1002/pro.70750","DOIUrl":"10.1002/pro.70750","url":null,"abstract":"<p><p>Glucagon is a well-established therapeutic peptide, widely used to treat hypoglycemia. Like many peptide drugs, it offers advantages such as specificity, biocompatibility, and high affinity for receptor targets, but suffers from limited physical and chemical stability. In particular, improper conditions can promote the formation of amyloid fibrils, leading to loss of biological activity and, in some cases, cytotoxicity. Understanding the conditions that modulate the structural behavior of glucagon is therefore crucial. Currently, two injectable formulations are available on the market: a lyophilized vial of glucagon with lactose at acidic pH, and a more recent auto-injector ready-to-use (RTU) formulation containing glucagon in dimethyl sulfoxide (DMSO) with trehalose. In this study, we investigated the conformational properties of glucagon in these two marketed formulations and compared them with glucagon dissolved in aqueous solution at pH 3.5, a metastable condition prone to aggregation. Preliminary circular dichroism and fluorescence spectroscopy were used to confirm glucagon stability over time; subsequently, NMR analysis showed that structural destabilization consistently begins at the C-terminal region, while the ¹¹Ser-Leu¹⁴ segment remains structured across all environments. These findings highlight two key determinants of glucagon stability and aggregation. Strategies that preserve the integrity of the C-terminal region while stabilizing the ¹¹Ser-Leu¹⁴ motif may improve peptide solubility and extend shelf life, providing a rational basis for the design of next-generation glucagon formulations for emergency use.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70750"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13443137/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148679794","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Systemic impact of intramuscular snake envenomation and intravenous immunoglobulin-based antivenom treatment on murine plasma proteome and peptidome.","authors":"Bianca C S C de Barros, Alison F A Chaves, Mariana S L C Morone, Ana T A Sachetto, Marcelo L Santoro, Oliver Schilling, Solange M T Serrano","doi":"10.1002/pro.70740","DOIUrl":"10.1002/pro.70740","url":null,"abstract":"<p><p>Snake envenomation remains a critical global public health challenge. In Brazil, the genus Bothrops is responsible for most accidents, typically resulting in coagulopathy, hemorrhage, and renal failure. Despite its clinical prevalence, the molecular mechanisms driving plasma protein disturbances remain poorly understood. In this study, we applied mass spectrometry to characterize the plasma proteome and peptidome of mice following a sub-lethal dose of Bothrops jararaca venom and subsequent single dose antivenom treatment at timepoints 3, 6 and 24 h. From a total of 781 identified proteins, quantification of 590 revealed systemic pathological signatures such as the lower abundance of key coagulation and complement proteins, indicating coagulopathy and immune disruption. Simultaneously, an increase in tissue damage markers, innate immune response proteins, and Damage-Associated Molecular Patterns was detected. In the acute phase, the biological response was highly conserved regardless of antivenom administration, while by 24 h the group treated only with venom exhibited prolonged inflammation and metabolic stress, whereas the venom + antivenom group transitioned to cellular recovery and detoxification. The peptidomic analysis identified a large degradome of 1964 unique peptides, showing proteolysis of fibrinogen, apolipoprotein A-I, complement factors and protease inhibitors, among others. The analysis of cleavage sites implicated the activity of cysteine-, metallo-, and serine proteases in this process. Our results demonstrate that B. jararaca venom induced a rapid, dramatic systemic imbalance and proteostasis disruption that was only partially mitigated by antivenom. These findings underscore the necessity of developing complementary therapies that target both the primary toxins and secondary host-driven pathology.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70740"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13442928/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148679785","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineered antibodies preserve structural and functional recognition of Plasmodium falciparum circumsporozoite protein.","authors":"Monika Jain, Sashank Agrawal, Fabien Cannac, Gonzalo E González-Páez, Wen-Hsin Lee, Re'em Moskovitz, Randal R Ketchem, Katherine L Williams, Daniel E Emerling, Andrew B Ward, Ian A Wilson","doi":"10.1002/pro.70738","DOIUrl":"10.1002/pro.70738","url":null,"abstract":"<p><p>Long-lasting and highly effective malaria vaccines or monoclonal antibodies (mAbs) remain urgently needed, as current interventions show age-dependent benefits rather than broad protection across all ages. Recently, two highly protective human mAbs 224 (IGHV3-49/IGLV1-40) and 7088 (IGHV3-33/IGKV1-5), encoded by distinct germline genes, were re-engineered and renamed MAM01 and MS-1805, respectively, to extend serum half-life and enable low-cost, large-scale manufacturing in alignment with WHO guidelines. MAM01 completed Phase I and IIb clinical trials (safety/PK/challenge) in US healthy naive adults and is now in Phase I trials (age-de-escalation studies) in Uganda. Here, we determined crystal structures of the antigen-binding fragments (Fabs) of engineered MAM01, MS-1805, and 7088 in complex with different regions of the Plasmodium falciparum circumsporozoite protein (PfCSP), including junctional, minor, and major repeat regions. Notably, Fab 7088 features an extended CDRL3 comprising 10 amino acids (CDRL3:10) instead of the canonical 8 amino acid CDRL3 (CDRL3:8) typically observed in V_H3-33/V_K1-5-encoded mAbs, indicating unique folding within its germline context. In addition, cryo-EM structures of Fabs 7088 and MS-1805, in complex with recombinant shortened CSP (rsCSP), revealed a regular spiral configuration stabilized by homotypic Fab-Fab interactions, whereas 224 and MAM01 did not form such assemblies. Structural comparisons demonstrated that engineered antibodies retained key molecular and hydrogen-bond interactions without significant conformational changes, thereby maintaining antigen recognition and preserving binding affinity. These findings demonstrate that targeted framework engineering can enhance therapeutic potential while preserving structural integrity and function, providing insights for next-generation antibody-based interventions against malaria.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70738"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13446027/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148679812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MAXTIA: A high-throughput platform for rapid functional epitope mapping by kinetic screening of mutant libraries.","authors":"Kihoon Kim, Ryo Matsunaga, Takanori Yokoo, Makoto Nakakido, Kouhei Tsumoto","doi":"10.1002/pro.70769","DOIUrl":"10.1002/pro.70769","url":null,"abstract":"<p><p>Accurate identification of epitope residues is essential for developing biopharmaceuticals and understanding the mechanisms of immune recognition. However, experimental approaches for residue-level epitope mapping remain time-consuming and labor-intensive, while accurate computational prediction of protein-protein interfaces remains challenging. Here, we present MAXTIA, a high-throughput kinetic screening platform that integrates cell-free protein synthesis with high-throughput surface plasmon resonance and demonstrate its application to alanine scanning-based functional epitope mapping. This workflow enables the rapid preparation and kinetic characterization of up to 384 protein variants, allowing the identification of functional epitope residues within 3 days while simultaneously providing binding affinity and kinetic parameters (K_D, k_on, and k_off). We applied MAXTIA to map the epitope of the single-domain antibody (VHH) N1 against the pentraxin domain of neuronal pentraxin-2 (NP2 PTX). Alanine substitutions that cause substantial affinity losses clustered within a localized region on the AlphaFold3-predicted NP2 PTX structure, defining a functional epitope site. These residues closely matched the interface observed in the NP2 PTX-VHH N1 crystal structure, validating the accuracy of MAXTIA. Beyond epitope identification, MAXTIA provides a simple and versatile platform for the quantitative analysis of protein-protein interactions, including high-throughput screening of antibody variants for affinity optimization. This approach should accelerate biopharmaceutical development and facilitate mechanistic studies of molecular recognition.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70769"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13479966/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148763092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Viral PDZ target sequences are not inherently more promiscuous than those derived from endogenous target proteins.","authors":"Cole D Masuga, Caroline Ceravolo, Elise F Tahti, Daniel A Pollard, Jeanine F Amacher","doi":"10.1002/pro.70772","DOIUrl":"10.1002/pro.70772","url":null,"abstract":"<p><p>PDZ domains are one of the largest families of short linear motif (or peptide) binding domains in the human proteome. These scaffolding domains are important in signaling and trafficking pathways, such as in the formation of tight junctions or in the postsynaptic density of neurons. PDZ domains are also targeted by several pathogenic viral proteins, including human papillomavirus (HPV), influenza, hepatitis, rabies, coronaviruses, among others. Previously, we investigated the specificity determinants of modulator, or non-motif, residues for two PDZ binding motifs (or PBMs) from the cystic fibrosis transmembrane conductance regulator (CFTR) and HPV16 E6 proteins to better understand differences in relative promiscuity between these similar sequences. To test whether viral PDZ-binding motifs are inherently more promiscuous than endogenous targets, we measured binding affinities for 7 endogenous and 7 viral PBMs across 8 well-studied Class I PDZ domains. Fluorescence anisotropy assays revealed no significant difference in relative binding affinities between viral and endogenous PBMs, indicating that viral versus endogenous origin alone does not explain PBM promiscuity. Our results were consistent with available data from high throughput holdup assays. Taken together, our data support previously reported sequence hotspots, whereby certain PBMs are recognized by large numbers of PDZ domains, likely due to a combination of widely favorable modulator residues.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70772"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13493693/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148797401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Facile, high-throughput protein purification enabled by high-efficiency protease elution.","authors":"Edson N Cárcamo Noriega, Ian S Truebridge, Frank D Teets, James W Bowman, Nathalia Rodriguez, Tessa A Howard, Christopher D Bahl","doi":"10.1002/pro.70746","DOIUrl":"10.1002/pro.70746","url":null,"abstract":"<p><p>While advances in artificial intelligence have made protein design widely accessible, protein production and characterization remain a bottleneck. Protease-cleavable affinity tags are commonly used to improve yield and purity of recombinant proteins, but tag removal adds labor and complexity. Previous attempts to use a selective protease to elute cleaved fusion protein during affinity chromatography have suffered from poor digestion efficiency that greatly diminishes yield, or protease contamination in the eluate. Here, we describe a novel SUMO protease construct that supports rapid, high-yield protease elution. This is the keystone of an end-to-end DNA-to-protein workflow that we optimized for speed, parallelizability, and generalizability. While our workflow is intended for an automated liquid-handler, it can easily be performed manually, making it broadly accessible. We demonstrate the method by producing and characterizing 96 disparate proteins derived from mesophilic organisms. This provides a benchmark for methods development and a curated dataset useful for machine learning.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70746"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13440354/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148679744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A cofactor-promiscuous HMGR from the Lyme disease pathogen illuminates diversity in bacterial isoprenoid biosynthesis.","authors":"Isaac A Paddy, Joshua McCausland, Madelyn Frazier, Poulami Chatterjee, Mekedlawit Setegne, Oliv Eidam, Christine Jacobs-Wagner, Laura M K Dassama","doi":"10.1002/pro.70766","DOIUrl":"10.1002/pro.70766","url":null,"abstract":"<p><p>The Lyme disease pathogen Borrelia burgdorferi contains a highly reduced genome lacking many primary metabolic pathways. However, B. burgdorferi retains the mevalonate pathway that synthesizes isopentenyl pyrophosphate (IPP), the precursor to the peptidoglycan carrier lipid. While the mevalonate pathway and the enzyme that catalyzes its rate-limiting step (3-hydroxy-3-methyl glutaryl coenzyme A reductase, HMGR) are well studied in vertebrates, little is known about the pathway in B. burgdorferi and many pathogenic bacteria. In this work, we reveal that HMGR is a critical metabolic enzyme in B. burgdorferi. We demonstrate that loss of HMGR causes morphological defects and muted de novo synthesis of peptidoglycan; these defects are ameliorated by exogenous mevalonate and IPP. Biochemical characterization unveiled HMGR as a highly unusual cofactor-promiscuous oxidoreductase that functions with both nicotinamide cofactors. Bioinformatics and biochemical characterization uncovered examples of similarly promiscuous HMGRs and revealed a previously unrecognized evolutionary link to cofactor choice. Moreover, structures of the enzyme reveal a highly divergent active site architecture. Together, these findings firmly establish HMGR as an opportunity target for the development of antibacterials for a diderm pathogen while highlighting cofactor promiscuity as an evolutionary acquired feature in HMGRs.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70766"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13488460/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148797374","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Peroxynitrite-induced structural remodeling and aggregation of hemoglobin under nitroxidative stress.","authors":"Shaik Basha, Aradhika Vijeev, Darshan Chikkanayakanahalli Mukunda, Krishna Kishore Mahato","doi":"10.1002/pro.70762","DOIUrl":"10.1002/pro.70762","url":null,"abstract":"<p><p>Peroxynitrite (PN), generated by the reaction of nitric oxide with superoxide, is a potent reactive nitrogen species (RNS) implicated in nitroxidative protein damage in cardiovascular and neurodegenerative diseases. Hemoglobin (Hb), the principal oxygen-transport protein of erythrocytes, is a key intravascular scavenger and target of PN; however, how controlled PN exposure translates into structural remodeling and aggregation of Hb remains poorly understood. Purified human Hb was exposed to PN across sub-stoichiometric to moderate oxidant excess and characterized using intrinsic fluorescence, UV-visible (UV-vis) spectroscopy, 2,4-dinitrophenylhydrazine (DNPH) carbonyl assay, Thioflavin T (ThT), 8-anilino-1-naphthalenesulfonic acid (ANS), Congo Red (CR), Fourier-transform infrared (FTIR) spectroscopy with Amide I deconvolution, dynamic light scattering (DLS) with zeta potential, x-ray powder diffraction (XRPD), scanning electron microscopy (SEM), atomic force microscopy (AFM), and fluorescence microscopy. At low oxidant loads, PN caused concentration-dependent fluorescence quenching, carbonyl accumulation, and graded UV-visible absorbance increases, consistent with heme-centered oxidation and limited aromatic residue modification. At higher concentrations, Amide I deconvolution revealed collapse of α-helical content from 66.41% to 8.81% and β-sheet enrichment to 71.18%, accompanied by surface charge neutralization, increased hydrodynamic diameter, and enhanced nanoscale roughness. Despite β-sheet accumulation and ThT/CR-positive aggregate formation, XRPD, AFM, SEM, and fluorescence microscopy showed no long-range crystalline order or fibrillar morphology, demonstrating that PN drives Hb toward amorphous and oligomeric assemblies rather than canonical amyloid fibrils. Time-dependent assays at 37°C confirmed that PN modification lowers the nucleation barrier for thermally driven aggregation. These findings define physicochemical thresholds separating protective scavenging from structural destabilization and offer a framework for distinguishing nitroxidative protein aggregation from classical amyloid fibrillation.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70762"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13463319/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148713503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A protein chaperone can stabilize the intein-containing precursor to indirectly promote protein splicing.","authors":"John S Smetana, Christopher R Powell, Noy Bagdadi, Tia M Ariagno, Hazel Thomas, Joel Weinberger, Eyal Gur, Christopher W Lennon","doi":"10.1002/pro.70768","DOIUrl":"10.1002/pro.70768","url":null,"abstract":"<p><p>Intervening proteins (inteins) interrupt host protein sequences and are removed by a self-mediated protein splicing reaction. Inteins are abundant in the microbial world and are often found within essential genes involved in DNA replication, recombination, and repair. Compelling examples of conditional protein splicing, where intein removal and subsequent host protein activation are highly dependent on environmental cues, suggest that some inteins serve as post-translational regulatory elements. Within the cellular context, however, the factors influencing protein splicing are still largely unknown. In this work, we demonstrate that the splicing of an intein from Mycobacterium smegmatis DnaB, an essential helicase, is temperature sensitive. Using two artificial extein systems-a kanamycin intein splicing reporter (KISR) and an in vitro splicing reporter-we show that splicing is inhibited at elevated, yet physiological, temperatures. In accordance with our results from M. smegmatis, we find that overexpression of GroEL in Escherichia coli expressing the KISR system drastically increased antibiotic resistance in a temperature-dependent manner. Under heat stress, we find that GroEL dramatically increases the levels of unspliced precursor protein. Mechanistically, GroEL does not appear to directly promote protein splicing, but rather increases the pool of unspliced precursor, which in turn results in more overall splicing. These findings represent the first description of a chaperone promoting protein splicing, demonstrating the ability of cellular factors to influence intein excision. From a physiological perspective, we hypothesize that a M. smegmatis DnaB intein may act as a switch to inhibit DNA replication under conditions of heat stress. Extending past this, our findings have broad implications for improving the efficiency of intein-based protein engineering technologies.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70768"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13476343/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148761942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of antibody binding by adjusting thermodynamic stability during the maturation of antibody affinities.","authors":"Yoko Akazawa-Ogawa, Tomonari Matsuda, Norihiko Kiyose, Nobuo Miyazaki, Tetsuo Fukuta, Motoyasu Adachi, Yuji Ito, Yoshihisa Hagihara","doi":"10.1002/pro.70739","DOIUrl":"10.1002/pro.70739","url":null,"abstract":"<p><p>Somatic mutations and antibody clone selection occur in B-cell hyper-evolution over extremely brief periods of time. Herein, we developed a technique for antibody screening by immunizing alpacas with antigens and observing antibody sequence transitions over time, a method we named Tracking the Evolution of Antibodies over time. This technique allows the observation of sequence transitions of somatic mutations in antibody populations originating from the same genome. The accumulation of somatic mutations was significantly associated with enhanced antigen-binding capacity and reduced stability in clusters. In comparison with the first clone to emerge in this cluster, numerous clones exhibited a decline in thermal stability, with a maximum variation of 21°C. Somatic mutations within the clusters demonstrating high similarity were observed to be concentrated in CDR-1, CDR-2, and FR-3. This suggests that these mutations have a significant impact on binding capacity and stability. However, the correlation between antigen binding capacity and stability was insignificant and weak. The thermal stability of antibodies correlated with acid and alkali resistance, with clones exhibiting lower thermal stability demonstrating higher acid and alkali resistance in antigen-antibody complexes. The results indicate that in the antibody selection process, the strength of antigen binding involves optimizing stability, with antibodies with more flexible structures being selected.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70739"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13445567/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148679722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}