Journal of Biological Engineering最新文献

{"title":"Ex vivo assay for organ-specific cancer cell invasion.","authors":"François Tyckaert, Paul Frieso Göddertz, Maria Reichhold, Bettina Sarg, Klaus Faserl, Pere Patón González, Felix Eichin, Andreas Villunger, Steffen Ormanns, Stefan Redl, Julia Hofmann, Theresa Hautz, Francesco Baschieri","doi":"10.1186/s13036-026-00719-9","DOIUrl":"10.1186/s13036-026-00719-9","url":null,"abstract":"<p><strong>Background: </strong>Metastasis is the leading cause of cancer-related mortality, yet experimental models often fail to recapitulate the tissue-specific microenvironments shaping metastatic dissemination. While in vivo systems provide physiological relevance, they remain challenging for mechanistic studies. Conversely, conventional in vitro assays lack the organ-specific extracellular matrix (ECM) that regulates invasive behavior. Accessible models that balance biological relevance with experimental feasibility are thus needed.</p><p><strong>Results: </strong>We developed an ex vivo invasion platform based on mild detergent decellularization of mouse organs followed by vibratome slicing. This approach generates optically transparent lung, liver, and intestine ECM scaffolds that preserve native matrix architecture, mechanical properties, and retain biochemical hallmarks of their tissues of origin. Organ-derived matrices were integrated into standard microfluidic channels and analyzed using conventional fluorescence microscopy to enable quantitative assessment of cancer cell invasion. Benchmarking with breast cancer cell lines of defined invasive capacity, we could demonstrate the robustness and biological relevance of the system. Non-invasive MCF7 cells failed to infiltrate any scaffold. In turn, highly invasive MDA-MB-231 cells successfully invaded permissive soils (lung/liver) but were unable to colonize the non-permissive soil (intestine). Our platform enabled quantitative assessment of invasion rates, and revealed organ-specific transcriptional programs associated with invasive adaptation by RNA-seq.</p><p><strong>Conclusions: </strong>The ex vivo organ-derived ECM framework presented here provides a scalable, cost-effective, and experimentally accessible system to study ECM-driven determinants of metastatic invasion. Preserving tissue-specific matrix cues while reducing reliance on animal models, it enables interrogation of ECM-driven metastasis mechanisms and therapeutic evaluation.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13632008/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148308108","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":"From coexistence to antagonism: nutrient- and temperature-dependent interactions between Bacillus atrophaeus JunSE1L and Pseudomonas chlororaphis O6 with implications for the wheat rhizosphere.","authors":"Anagha Wankhade, Ashlynn Clark, David W Britt","doi":"10.1186/s13036-026-00716-y","DOIUrl":"10.1186/s13036-026-00716-y","url":null,"abstract":"<p><strong>Background: </strong>Interactions between beneficial plant-associated microbes influence microbial community structure and plant health, yet antagonistic relationships among co-occurring beneficial bacteria remain poorly understood. This study examined interactions between Pseudomonas chlororaphis O6 (PcO6), a wheat root-colonizing epiphyte, and JunSE1L, a Bacillus atrophaeus seed-borne endophyte isolated from winter wheat (Triticum aestivum var. Juniper).</p><p><strong>Results: </strong>For wheat grown in a sterile sand matrix, JunSE1L emerged from seeds and colonized roots; however, when PcO6 was introduced at the bottom of the growth boxes, it migrated and became the dominant rhizoplane colonizer. JunSE1L emerged from both damaged roots and shoots, and produced biosurfactants that lowered the surface tension of water to 30 mN/m. On agar plates at 22 °C PcO6 strongly inhibited JunSE1L growth on minimal medium (MM) and to a limited extent on rich LB medium; inhibition was absent on both media at 37 °C. On MM, JunSE1L colonies adopted dendritic, highly spread morphologies, potentially from biosurfactant release that may have also facilitated PcO6 swarming over established JunSE1L colonies. Whole PcO6 supernatant and a < 3 kDa filtrate each suppressed JunSE1L growth in liquid MM at 22 ˚C. The inhibitory compound(s) was resistant to proteinase K digestion and heat deactivation, exhibiting bacteriostatic activity against JunSE1L.</p><p><strong>Conclusions: </strong>PcO6 inhibits JunSE1L in a temperature-, nutrient-, and density-dependent manner: On rich growth media they coexist, while on defined minimal media, PcO6 dominates, suggesting that resource limitation intensifies competitive interactions and favors epiphyte (PcO6) - mediated suppression of endophyte (JunSE1L). While both bacteria comprise the wheat root microbiome, under conditions recapitulating the rhizosphere (defined nutrients, moderate temperature, and PcO6 already present in the soil) PcO6 release of a bacteriostatic agent may repress JunSE1L emergence. To the contrary, JunSE1L release of biosurfactants may facilitate PcO6 migration to the rhizoplane.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13540836/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148301676","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":"Research progress and applications of functional hydrogels in nasal wound healing.","authors":"Chongyang Wang, Zhikai Wang, Shengqi Gan, Bin He, Yejingwen Tong, Yaojie Zhu, Dong Ye","doi":"10.1186/s13036-026-00721-1","DOIUrl":"10.1186/s13036-026-00721-1","url":null,"abstract":"<p><p>Nasal wound healing presents a distinct clinical challenge in otorhinolaryngology, complicated by the unique physicochemical environment of the nasal cavity-including a slightly acidic mucosal pH (≈ 6.3-6.9), shear-thinning mucus rheology (healthy viscosity ≈ 0.1-1 Pa·s, elevated to 1.6-3.2 Pa·s in chronic sinusitis), continuous mucociliary clearance (MCC) with ciliary beat frequencies of ~ 10 Hz, and enzymatic activity dominated by lysozyme. Conventional nasal packing materials-such as Vaseline gauze and polyvinyl alcohol sponges-suffer from poor biocompatibility, rigid mechanical properties, and traumatic removal, failing to satisfy these demanding physicochemical constraints. Functional hydrogels, owing to their tunable viscoelasticity, dynamic covalent or supramolecular crosslinking, and intrinsic responsiveness to pH, reactive oxygen species (ROS), temperature, and enzymes, have emerged as a next-generation solution. This review systematically profiles the physicochemical microenvironment of the nasal wound; reclassifies hydrogel systems by polymer backbone (natural polysaccharides/proteins vs. synthetic polymers) and crosslinking chemistry (dynamic covalent bonds: Schiff base, disulfide, boronic ester vs. supramolecular: host-guest, hydrogen bonding); and examines reaction pathways, degradation kinetics, and mechanical matching with nasal mucosa. Representative systems are critically evaluated, including: (i) the RADA16 self-assembling peptide hydrogel, which in randomized clinical trials reduced postoperative adhesion by 91-100% compared with gelatin-thrombin and bioresorbable controls; (ii) TSPBA/PVA ROS-responsive hydrogels that scavenge H₂O₂ via boronic ester cleavage (second-order rate constant ≈ 0.1-1 M⁻¹·s⁻¹), prolonging mucosal retention to 24 h; (iii) pNIPAAm-based thermoresponsive systems with LCST ≈ 32 °C, the critical implications of which for the physiological nasal temperature range (30-34 °C) are discussed. We further analyze cell-material interactions governing macrophage M1→M2 polarization, epithelial/fibroblast behavior, and ciliary function. Ongoing clinical trials (NCT04970966, NCT05898074) and persistent translational challenges-including sterilization, long-term in vivo safety, and standardized large-scale production-are critically addressed, offering a materials-science-focused framework for the design of next-generation nasal hydrogels.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13540062/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148301750","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":"Engineering of Corynebacterium glutamicum for biosynthesis of the pharmaceutically active N-acetyltyramine: establishing and optimizing de novo production.","authors":"Sara-Sophie Poethe, Lilli Sophie Kaimann, Kai H Schülke, Stephan C Hammer, Volker F Wendisch","doi":"10.1186/s13036-026-00713-1","DOIUrl":"10.1186/s13036-026-00713-1","url":null,"abstract":"<p><strong>Background: </strong>N-Acetyltyramine is increasingly recognized for its diverse pharmaceutically relevant properties. In particular, it exhibits antimicrobial activity against multi-drug resistant pathogens, which pose a considerable health challenge. Therefore, the demand for a sustainable and efficient N-acetyltyramine production arises.</p><p><strong>Results: </strong>Here, we engineered Corynebacterium glutamicum for de novo production of N-acetyltyramine. Heterologous overexpression of different bacterial and insect acetyltransferase genes in a tyramine overproducing strain identified the arylalkylamine N-acetyltransferase from Bombyx mori as the most promising enzyme, resulting in an N-acetyltyramine titer of 5.5 ± 0.3 mM. Design of Experiment-based optimization of the culture medium revealed urea and L-phenylalanine as media components significantly affecting N-acetyltyramine production. Combining media optimization with genetic engineering to balance the expression strength of the genes encoding the L-tyrosine decarboxylase from Levilactobacillus brevis and the arylalkylamine N-acetyltransferase from B. mori increased de novo N-acetyltyramine production to 13.8 ± 0.1 mM (2.5 ± 0.1 g/L). The optimized medium composition was also transferable to the production of other L-tyrosine derivatives, tyramine and tyrosol, increasing their titers by 24% and 44%, respectively. In addition, supplementing the established N-acetyltyramine producing strain with propionate as a second carbon source enabled the production of N-propionyltyramine. To the best of our knowledge, this represents the first report describing biotechnological production of N-propionyltyramine.</p><p><strong>Conclusion: </strong>In this study, de novo N-acetyltyramine production was established and substantially increased through the combined implementation of metabolic engineering and media optimization strategies. The successful transfer of the optimized medium to enhance the production of additional L-tyrosine derivatives underscores the potential of integrating genetic engineering with culture media refinement to improve biotechnological processes.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"20 1","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13281258/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148277439","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":"Engineering Fc heterodimerization via asymmetric β-strand reconfiguration for bispecific antibody assembly.","authors":"Arkaitz Cano, Izaskun Morillo, Ainhoa Goenaga, María Elena Laugieri, Ander de Blas, Eunate Valdaliso-Díez, June Ereño-Orbea, Beatriz Apellaniz, Edurne Rujas","doi":"10.1186/s13036-026-00717-x","DOIUrl":"10.1186/s13036-026-00717-x","url":null,"abstract":"<p><p>Bispecific antibodies (bsAbs) have unlocked new therapeutic modalities by enabling dual antigen targeting, offering transformative potential in oncology and autoimmune diseases. However, the efficient production of full-length IgG-like bsAbs remains hindered by mispairing of heavy and light chains, yielding heterogeneous, low-purity products. Here, we present an Fc engineering strategy, termed StrandLock, that enables the controlled assembly of bsAbs through asymmetric β-strand redistribution at the CH3 dimer interface, in which the β-strand segment (residues N390-S400) is duplicated in one heavy chain and deleted from the partner. This structural asymmetry, combined with targeted point mutations, promotes obligate heterodimerization while suppressing homodimer formation. Importantly, these modifications enable the correct assembly of bsAbs with purities approaching 99% and allow streamlined manufacturing via a single-step affinity purification process comparable to that used for monoclonal antibodies. The engineered bsAbs retain key Fc effector functions and exhibit comparable in vivo pharmacokinetics to native IgG. Structural analysis confirms preservation of the CH3 fold and native-like interfacial packing. Despite a modest reduction in thermal stability, the molecules remain aggregation-resistant and functionally robust under accelerated stress conditions. We further demonstrate the versatility of the StrandLock design across multiple antibody pairs underscoring its utility as a strategy for bsAb production.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13520406/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148277497","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":"Neuroprotective effects of C-phycocyanin and allophycocyanin from Spirulina purified using a newly developed method: in vitro barrier permeability, antioxidant and anti-inflammatory activities.","authors":"Antonella Girgenti, Pasquale Picone, Laura Palumbo, Rosaria Lauceri, Domenico Nuzzo","doi":"10.1186/s13036-026-00720-2","DOIUrl":"10.1186/s13036-026-00720-2","url":null,"abstract":"<p><p>Phycobiliproteins, particularly C-phycocyanin (C-PC) and allophycocyanin (APC), are major bioactive pigments in Arthrospira platensis (Spirulina) with well-documented antioxidant properties, yet their individual neuroprotective potential and bioavailability remain poorly characterized. Here, we developed an efficient, potentially scalable, two-step ultrasound-based extraction followed by membrane chromatography to obtain analytical-grade purity C-PC and APC from fresh A. platensis biomass. Both purified phycobiliproteins, together with the crude extract (CE), demonstrated relevant cell viability up to 35 µg/mL in SH-SY5Y neuronal and BV2 microglial cells. In vitro assays revealed that APC exhibited significantly higher ferric-reducing antioxidant power (FRAP) than C-PC, while both phycobiliproteins effectively reduced intracellular ROS levels and protected SH-SY5Y cells from tert-butyl hydroperoxide-induced oxidative stress, with C-PC and APC showing the strongest cytoprotective effects. All preparations also significantly decreased LPS-induced nitric oxide production in BV2 microglia, indicating anti-inflammatory activity. Using Caco-2 monolayers, we demonstrated for the first time that both purified C-PC and APC exhibit time-dependent, low but moderate transcellular permeability, while preserving monolayer integrity. Fluorescence microscopy confirmed substantial intracellular uptake in SH-SY5Y neurons (punctate cytoplasmic pattern) and lower, yet detectable, internalization in BV2 microglia. These findings highlight distinct yet complementary bioactivities of highly purified C-PC and APC and provide the first evidence of their ability to cross physiological barriers in vitro, offering new mechanistic insights into their potential as neuroprotective nutraceuticals targeting oxidative and neuroinflammatory processes in the central nervous system.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13520510/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148271554","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":"Systems-level analysis identifies IRF6 as an inhibitor of epithelial-mesenchymal transition.","authors":"Ayalur Raghu Subbalakshmi, Aditya Agrawal, Shibjyoti Debnath, Kishore Hari, Sarthak Sahoo, Jason A Somarelli, Mohit Kumar Jolly","doi":"10.1186/s13036-026-00706-0","DOIUrl":"10.1186/s13036-026-00706-0","url":null,"abstract":"<p><strong>Background: </strong>Epithelial-mesenchymal transition (EMT) and its reverse process mesenchymal-epithelial transition (MET) are crucial during metastasis and therapy resistance. While the dynamics and master regulators of EMT are well-studied, the transcription factors that can prevent EMT or promote MET are relatively less understood.</p><p><strong>Results: </strong>Here, by integrating bulk and spatial transcriptomic data analysis from cell lines and patient samples with mechanism-based dynamical modelling we identify IRF6 as a factor that strongly associates with an epithelial phenotype and is often inhibited during EMT. In vitro experiments in multiple cancer cell lines demonstrate the progression to a mesenchymal phenotype upon IRF6 knockdown, suggesting a role as an inhibitor of EMT. Finally, we observe that IRF6 expression levels correlate with worse patient survival in a subset of solid tumour types.</p><p><strong>Conclusion: </strong>Our integrated computational-experimental, systems-level analysis suggests that IRF6 is frequently downregulated during EMT and can also prevent the progression towards a complete EMT, underscoring its role as an MET stabilizing factor.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13491704/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148251723","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":"Unravelling the metabolism of itaconate and related branched C5-dicarboxylates for CO₂-based production in Cupriavidus necator H16.","authors":"Erik K R Hanko, Naglis Malys","doi":"10.1186/s13036-026-00711-3","DOIUrl":"10.1186/s13036-026-00711-3","url":null,"abstract":"<p><p>Branched C5-dicarboxylates (C5-DCAs), such as itaconate, are high-value platform chemicals with diverse applications in the synthesis of bio-based polymers. In addition to their industrial relevance, itaconate has attracted attention as a key immunomodulatory metabolite. In this study, we demonstrated that Cupriavidus necator can grow on itaconate as its sole carbon source and identified key genes involved in its degradation through targeted deletions. These include multiple functional homologs of itaconate CoA-transferase, itaconyl-CoA hydratase, and (S)-citramalyl-CoA lyase, which collectively mediate itaconate catabolism. We also identified ItcR1, a LysR-type transcriptional regulator, as a key mediator of itaconate-responsive gene expression. Transcriptomic analysis revealed broader transcriptional responses to itaconate, methylsuccinate, and mesaconate, suggesting an expanded role of C5-DCA metabolism. Finally, by expressing a heterologous cis-aconitate decarboxylase, we achieved the autotrophic production of a C5-DCA mixture of methylsuccinate, citramalate, mesaconate, and itaconate from CO₂ and hydrogen. These findings provide new insight into branched C5-DCA metabolism in C. necator and advance its development as a chassis for the sustainable production of these industrially relevant compounds from inorganic feedstocks.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13491972/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148251695","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":"MelC tyrosinase-dependent enzymatic synthesis of functional melanins from phenolic monomers.","authors":"Nimya Krishnan, Koijam Monica Devi, Chan-Seo Yeo, Kwon-Young Choi","doi":"10.1186/s13036-026-00710-4","DOIUrl":"10.1186/s13036-026-00710-4","url":null,"abstract":"<p><p>Sustainable microbial bioproduction offers a promising route to synthesize high-value melanin biopolymers, yet its scalability is fundamentally limited by the high cost of the L-tyrosine precursor. Here, we aimed to overcome this economic barrier by a highly efficient, engineered microbial platform that utilizes recombinant Escherichia coli expressing MelC tyrosinase to convert diverse, cost-effective noncanonical phenolic monomers into customizable melanin-like biopolymers. Among 13 screened substrates, p-Cresol (PC) and 4-hydroxyphenylacetic acid were the most efficient precursors, yielding 2.40 g/L and 3.92 g/L, respectively. Structural analyses (Ultraviolet-visible spectroscopy, Fourier-transform infrared, Solid-state NMR, MALDI-TOF MS, scanning electron microscopy, Energy-dispersive X-ray spectroscopy, X-ray diffraction, EPR) revealed that these biopolymers possess a native eumelanin-like amorphous architecture while exhibiting partial Dimethyl sulfoxide solubility for improved processability. The synthesized eumelanin-like pigments exhibited high thermal stability (retaining > 37% mass at 800 °C) and strong antioxidant activity, with PC-melanin outperforming conventional eumelanin by twenty-fold. Most importantly, these novel substrates are effluents of industrial and agriculture waste. Thus, this strategy frames a new method for bioremediation. These customizable eumelanin-like pigments hold strong potential for demanding industrial applications, including high-temperature coatings and advanced electronics.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13480188/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148217860","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":"Engineering a conditionally active cetuximab prodrug via affibody-based paratope masking.","authors":"Anna Mestre Borras, Hanna Mehari, Athanasios Bitzios, Ivan Zelepukin, Moeen Ud-Din, Tianqi Xu, Sihan Li, Anna Orlova, Vladimir Tolmachev, Stefan Ståhl, Anzhelika Vorobyeva, John Löfblom","doi":"10.1186/s13036-026-00705-1","DOIUrl":"10.1186/s13036-026-00705-1","url":null,"abstract":"<p><p>The application of monoclonal antibodies as targeted drugs has revolutionized cancer therapy, yet their efficacy is sometimes limited by toxicities arising from target expression in healthy tissues. Various strategies have emerged to overcome these issues, including the design of antibody-based prodrugs. Here, we report the generation of affibody-based masking domains designed to specifically bind and mask the paratope of the anti-epidermal growth factor receptor (EGFR) antibody, cetuximab. Bacterial display of affibody library and cell sorting were employed to isolate suitable candidates. Both experimental and computational analyses confirmed that several of these selected affibodies effectively blocked cetuximab from binding EGFR. A cetuximab prodrug was created by fusing the most promising masking candidate to the antibody heavy chains. The prodrug demonstrated over 400-fold difference in growth inhibition effect between its masked and unmasked states in vitro. A biodistribution study comparing cetuximab and its prodrug demonstrated the feasibility of conditional activation of the cetuximab prodrug in a xenograft mouse model. This study is the first to describe the use of an affibody masking domain to create an antibody-based prodrug, demonstrating significant potential for further development of safer and more efficacious cancer therapies.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"20 1","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13248396/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148205038","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}