AMB Express最新文献

{"title":"Bioactive screening and molecular docking of phenolics from Stellaria pallida.","authors":"Mohamed T Selim, Mohamed E Elnosary, Fatma Sh Abd El-Gwaid","doi":"10.1186/s13568-026-02090-w","DOIUrl":"10.1186/s13568-026-02090-w","url":null,"abstract":"<p><p>Stellaria pallida L. (Dumort.) is a species that belongs to the Caryophyllaceae family and has a broad range of secondary metabolites and therapeutic potential. In this study, the aerial parts of Stellaria pallida L. (S. pallida) were extracted by various organic solvents, viz. Chloroform, ethyl acetate, ethanol, and methanol, among others, were analyzed by GC/MS and HPLC-DAD to determine their chemical composition and antimicrobial properties. In addition, an in silico molecular docking analysis by AutoDock Vina was performed to evaluate the potential interactions between the identified phytochemical compounds and selected antimicrobial target proteins. The methanol extract had the maximum total phenolic (58±1.55 mg GAE/g), and the ethanol extract recorded the maximum total values of flavonoid (27.22±0.88 mg QE/g). Phytol, stigmasterol, Neophytadiene, rutin, chlorogenic acid, ferulic acid, apigenin-7-glucoside, and epicatechin were the major chemical compounds detected in the S. pallida plant. Additionally, the methanolic extract of S. pallida possesses more antioxidant activity in both DPPH and H₂O₂ scavenging assays, with IC₅₀ values of 16.24 and 13.54 µg/mL, respectively, compared to other solvent extracts. The antimicrobial evaluation using the agar well diffusion method further confirmed the superior efficacy of the methanolic extract, showing the lowest MIC values against C. albicans (6.25 mg/mL), followed in order by S. aureus and B. subtilis (12.5 mg/mL), while E. coli and K. pneumoniae required higher concentrations (25 mg/mL). Furthermore, molecular docking results revealed strong binding affinities of several identified compounds, particularly rutin and chlorogenic acid, with key antimicrobial target proteins, supporting the observed biological activities. These findings highlight the methanolic extract of S. pallida as a reliable source of natural antioxidants and broad-spectrum antimicrobial activity, in addition to its utility against numerous infectious diseases.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":"16 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13569792/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148944109","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":"Juniperus phoenicea extract-loaded nanostructured lipid carriers for potent antimicrobial and anticancer combination therapy.","authors":"Passant A Ismail, Bassma H Elwakil, Mohamed Hagar, Zakia A Olama","doi":"10.1186/s13568-026-02054-0","DOIUrl":"10.1186/s13568-026-02054-0","url":null,"abstract":"<p><p>This study evaluates the antimicrobial properties of Juniperus phoenicea L. against multidrug-resistant microbes, highlighting potential synergistic effects of combining the J. phoenicea L. newly synthesized nanoparticles with conventional antibiotics and anticancer agents. Juniperus phoenicea L. ethanolic and ethyl acetate extracts were prepared. Antibacterial activity against a panel of multidrug-resistant microbes showed the ethanolic extract as superior to the ethyl acetate extract, with Acinetobacter baumannii being the most susceptible organism (minimum inhibitory MIC = 12.5 µg/mL). Phytochemical profiling of the ethanolic extract revealed predominant components, namely: Apigenin-7-glucoside, α-pinene, and Cedrol. Network pharmacology-based analysis suggested the potent effect of these compounds as anticancer and antibacterial agents. J. phoenicea L. nanoparticles were synthesized by a combined hot-melt dispersion and homogenization approach. Characterization yielded a zeta vesicle size of 98.6 nm, PDI 0.23, zeta potential + 48.5 mV, EE% 92.4%, and TEM size 76.2 nm. The nanoparticles markedly enhanced antibacterial activity (MIC = 6.25 µg/mL) with complete eradication of A. baumannii after 16 h. The combination with Amoxicillin and Cefoxitin showed synergistic effects. In anticancer assays, nanoparticles outperformed Fluorouracil, with IC₅₀ = 9.05 ± 0.49 µg/mL (≈ 8.7-fold higher anticancer potency than the crude extract). The Fluorouracil-nanoparticle combination further reduced IC₅₀ to 6.12 ± 0.41 µg/mL. Hence, it was concluded that J. phoenicea L. ethanolic extract exhibits potent antibacterial and anticancer properties and provides an effective nanoparticle precursor. Synergistic interactions with antibiotics and Fluorouracil suggest a multifaceted therapeutic strategy against infections and cancer, meriting further in vivo studies and mechanistic exploration.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":"16 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13550337/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148904928","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":"Retraction Note: Maslinic acid induces anticancer effects in human neuroblastoma cells mediated via apoptosis induction and caspase activation, inhibition of cell migration and invasion and targeting MAPK/ERK signaling pathway.","authors":"Yusheng Liu, Hongting Lu, Qian Dong, Xiwei Hao, Lingyan Qiao","doi":"10.1186/s13568-026-02107-4","DOIUrl":"10.1186/s13568-026-02107-4","url":null,"abstract":"","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":"16 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13507011/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148817122","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 novel synergistic phycobiliprotein combination against HIV-1 shows potent antiviral activity.","authors":"Pratiksha Jadaun, R Harshithkumar, Ishrat Khan, Debashree Dass, Pooja Wakchaure, Ruchi Kadam, Ashwini More, Kishore Dhotre, Nikhat J Siddiqi, Ahmed Alsadoun, Abdul Malik, Shyam Sundar Nandi, Vijay Nema, Abdul Arif Khan, Anupam Mukherjee","doi":"10.1186/s13568-026-02100-x","DOIUrl":"https://doi.org/10.1186/s13568-026-02100-x","url":null,"abstract":"<p><p>Human immunodeficiency virus type 1 (HIV-1) remains a major global public health challenge due to lifelong persistence, latency, and the emergence of drug resistance. Natural bioactive compounds are increasingly being explored as alternative or adjunct antiviral strategies. In this study, we evaluated the antiviral efficacy and cytotoxicity of a synergistic pharmaceutical composition comprising three phycobiliproteins: C-phycocyanin (C-PC), B-phycoerythrin (B-PE), and allophycocyanin (APC), derived from cyanobacteria and marine algae. Individual and combinatorial effects were assessed against HIV-1 using TZM-bl cells and peripheral blood mononuclear cells (PBMCs). A Box-Behnken experimental design was employed to identify optimal synergistic ratios. Among fifteen combinations initially screened at a fixed concentration of 50 µg/mL, five exhibited strong antiviral activity. The optimized formulation (CBA: C2; 73.28% C-PC, 20.70% B-PE, and 6.02% APC) produced up to ~ 100% inhibition of HIV-1 during the screening assay. Subsequent dose-response analyses demonstrated IC₅₀ values ranging from 5.01 to 12.60 µg/mL across different antiviral assays, while exhibiting minimal cytotoxicity (CC₅₀: 577.56 ± 34.5 µg/mL in TZM-bl cells and 247.64 ± 25.49 µg/mL in PBMCs). The formulation showed a high selectivity index, indicating a favourable therapeutic window. These findings highlight the potential of phycobiliprotein-based synergistic formulations as promising candidates for the development of safe and effective anti-HIV-1 therapeutics.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":"16 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13631182/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149102869","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":"Probiotic and safety characterization of Lactiplantibacillus plantarum ZZM01 isolated from mink.","authors":"Yao Wu, Cong Huang, Jianhua Wu, Yalin Zhou, Keyuan Liu, Weili Sun","doi":"10.1186/s13568-026-02088-4","DOIUrl":"https://doi.org/10.1186/s13568-026-02088-4","url":null,"abstract":"<p><p>In recent years, probiotics have attracted increasing attention due to their roles in gut health regulation, antimicrobial activity, and metabolic benefits. Given the high-fat dietary characteristics of mink, their intestinal microbiota represents a potential source of functionally valuable lactic acid bacteria. In this study, we evaluated the probiotic properties, safety profile, and functional potential of Lactiplantibacillus plantarum ZZM01 isolated from mink intestine. ZZM01 exhibited good tolerance to simulated gastrointestinal conditions, with survival rates of 71.92% at pH 2.0 and 84.71% in 0.3% bile salts. The strain showed inhibitory activity against several pathogenic bacteria, no hemolytic activity, and susceptibility to ceftriaxone and chloramphenicol. Its auto-aggregation ability increased over time (up to 93.85% at 24 h), and the surface hydrophobicity reached 33.19%. Additionally, ZZM01 displayed strong antioxidant activity, with a DPPH radical scavenging rate of 84.62%. In vitro cholesterol removal reached 44.94%, although no bile salt hydrolase activity was detected. Whole-genome analysis revealed multiple genes associated with stress tolerance, including acid, bile salt, oxidative, osmotic, heat, and cold stress responses, which were consistent with the observed gastrointestinal tolerance phenotype. Genome annotation also identified genes involved in carbohydrate metabolism, amino acid biosynthesis, and putative bacteriocin production. Safety-related genome mining identified multiple antibiotic resistance-associated genes and putative virulence factor-related genes, while no toxin- or invasion-associated genes were detected. Overall, ZZM01 exhibits strong gastrointestinal tolerance, antimicrobial activity, antioxidant capacity, adhesion-related properties, and favorable genomic characteristics, supporting its potential application as a probiotic strain.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":"16 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149226890","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigation of the antimicrobial, antibiofilm, and immunomodulatory activities of cranberry (Vaccinium macrocarpon L.) proanthocyanidins against multidrug-resistant pathogens.","authors":"Mai Zafer, Omar Zayan, Rawan Nasser, Nadeen Hatem, Mohamed Al Tamawi, Dalia Hamdy, Sayed E El-Sayed","doi":"10.1186/s13568-026-02092-8","DOIUrl":"10.1186/s13568-026-02092-8","url":null,"abstract":"<p><p>The increasing emergence of multidrug-resistant (MDR) pathogens poses a significant challenge to global healthcare, necessitating the development of alternative antimicrobial strategies. In the present study, cranberry extract was evaluated for its antimicrobial, antibiofilm, antioxidant, and anti-inflammatory activities against a panel of clinically relevant microorganisms, including Escherichia coli, Acinetobacter baumannii, Klebsiella pneumoniae, Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus aureus (MRSA). The extract demonstrated notable antimicrobial activity, with minimum inhibitory concentration (MIC) values ranging from 32 to 512 µg/mL, exhibiting relatively higher efficacy against Gram-positive bacteria. Significant antibiofilm activity was observed at sub-inhibitory concentrations, where cranberry extract reduced biofilm formation by up to 75-80% at 1/2 MIC and 50-65% at 1/4 MIC (p < 0.05) across tested isolates. Scanning electron microscopy (SEM) further supported these findings by revealing alterations in biofilm architecture following treatment. In addition, the extract exhibited strong antioxidant activity, achieving more than 85% free radical scavenging activity at higher concentrations. Cytotoxicity assessment revealed an IC₅₀ of approximately 450-500 µg/mL, with selectivity indices indicating a favorable therapeutic window, particularly against Gram-positive bacteria. Furthermore, in vivo evaluation using an infection model demonstrated improved survival rates (> 90%) and significant reductions in pro-inflammatory markers, including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), nuclear factor kappa B (NF-κB), and cyclooxygenase-2 (COX-2), compared with untreated infected controls (p < 0.05). Collectively, these findings highlight the potential of cranberry extract as a multifunctional therapeutic agent with antimicrobial, antibiofilm, antioxidant, and anti-inflammatory properties. The observed efficacy against MDR pathogens suggests its potential application as a complementary or alternative strategy for the management of resistant infections.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":"16 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-07-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13385296/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148519875","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":"Microbial functions in skin inflammation: a Mendelian randomization study.","authors":"Yujie Yang, Yiyuan Zhong, Peng Wu, MingXuan Li, Jingyi Cai, Qihong Chen, Xuwei Zheng, Haiming Chen, Teng Yu","doi":"10.1186/s13568-026-02094-6","DOIUrl":"10.1186/s13568-026-02094-6","url":null,"abstract":"<p><p>Inflammatory skin diseases (ISDs) represent a significant global health burden, yet the role of the human microbiome in their pathogenesis remains unclear. This study employed a comprehensive two-sample Mendelian randomization (MR) framework to investigate potential causal associations between multi-site microbiota (3117 oral, 412 gut, and 150 skin microbial features) and five major ISDs: psoriasis, atopic dermatitis, acne, seborrheic dermatitis, and urticaria. Genetic instruments were selected from large-scale genome-wide association studies (GWAS) of microbiota composition and ISD outcomes. The inverse variance weighted (IVW) method served as the primary analytical approach, supplemented by MR-Egger regression, weighted median, and MR-PRESSO for sensitivity analyses. Multiple testing was controlled using the false discovery rate (FDR) method. Our analyses identified numerous suggestive associations between specific microbial taxa, metabolic pathways, and ISD risk. Notably, shared microbial signatures were observed across multiple ISDs, suggesting potentially shared microbiome-associated pathogenic pathways. These results provide genetic evidence supporting a role for the microbiome in ISD susceptibility and highlight potential microbial targets for future therapeutic development.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13638966/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148468292","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":"Solid-state fermentation of tropical agro-industrial wastes using Pleurotus ostreatus for fungal protein production.","authors":"Kingsley I Eneogwe","doi":"10.1186/s13568-026-02091-9","DOIUrl":"10.1186/s13568-026-02091-9","url":null,"abstract":"<p><p>Solid-state fermentation (SSF) of lignocellulosic agro-industrial wastes using white-rot fungi represents a scalable, low-energy strategy for converting underutilised tropical biomass into high-quality fungal protein. This study optimised six substrate formulations combining cocoa pod husk (CPH), hardwood sawdust, and composted vegetable waste for Pleurotus ostreatus NIHORT EM-1 (ITS rDNA authenticated; GenBank MN165852; 99.8% identity to CBS 244.31) under Nigerian tropical conditions. The strain is deposited at the National Horticultural Research Institute (NIHORT), Ibadan, Nigeria, and is publicly available for research purposes upon written request to the Director of Research, NIHORT, P.M.B. 5432, Idi-Ishin, Ibadan, Nigeria (email: info@nihort.gov.ng; Tel: +234-803-000-0001; online germplasm catalogue: www.nihort.gov.ng/germplasm ). Substrate carbon-to-nitrogen (C:N) ratio, mycelial colonisation kinetics, fresh-weight yield, proximate nutritional quality (protein digestibility-corrected amino acid score, PDCAAS), and contamination incidence were evaluated across a completely randomised design (n = 5 replicates per treatment; 30 bags total). The optimal formulation [T5: 58% sawdust, 25% CPH, 10% composted vegetable waste, 5% rice bran; C:N 31.2:1] achieved biological efficiency (BE) of 116.9%, protein yield of 359 g kg⁻¹ dry substrate (PDCAAS = 0.87; in vitro digestibility 89.2%), and full mycelial colonisation in 18.4 ± 1.0 days - improvements of 57%, 126%, and 21%, respectively, over the sawdust-only control. T5 fruiting bodies contained 30.7 ± 1.8% crude protein (dry weight), with eight of nine essential amino acids exceeding WHO/FAO (2007) adult reference requirements. ISO 14040-compliant life cycle assessment confirmed 98-99% reductions in global warming potential, water use, and land use relative to beef protein. Enterprise modelling demonstrated a return on investment of 321% per 100 kg production cycle (Monte Carlo-confirmed; n = 10,000 iterations). Multi-site pilot validation across three contrasting Nigerian agro-ecological zones yielded 95.7% mean yield retention (ANOVA: F(2,27) = 2.14, p = 0.136), confirming protocol transferability. These results establish CPH-based SSF as a globally competitive, low-carbon fungal protein platform with direct relevance to agro-waste valorisation and food security in tropical West Africa.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13631136/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148403187","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":"Combination antifungal therapy against resistant C. albicans: efficacy and immune response in mice.","authors":"Ali Taghizadeh-Maleki, Farahnaz Hatami, Sima-Sadat Seyedjavadi, Farnoush Asghari-Paskiabi, Zahra Jahanshiri","doi":"10.1186/s13568-026-02085-7","DOIUrl":"10.1186/s13568-026-02085-7","url":null,"abstract":"<p><p>This study demonstrates the therapeutic effects of combination therapies against Candida albicans in mouse models, as opposed to mono-therapeutic treatments, while also comparing the pathogenicity of both sensitive and resistant strains. Fluconazole-resistant strain of Candida albicans was isolated from an infected patient, and its infectivity potential was compared with a susceptible strain by injecting a fungal cell suspension into the mice model. Additionally, the level of secretory interleukin 17 in the mice was examined. Furthermore, the efficacy of combination therapy was assessed using Fluconazole-Caspofungin (Flu-Cas) and Fluconazole-Amphotericin B (Flu-AmB) in comparison to monotherapy. The results indicate that the resistant strain exhibits higher pathogenicity compared to the susceptible strain in inducing systemic infection within the mouse model, as significant differences between the resistant and susceptible groups were observed at day 5 (p = 0.0401) and day 7 (p = 0.0182). The concentrations of IL-17 (pg/ml) on days 1 to 7 were 43.66 to 259 pg/ml, and 39.66 to 210 pg/ml in resistant and susceptible groups, respectively, which were only significant on days 5 (p = 0.0356) and 7 (p = 0.0280). Combination therapies using Flu-Cas and Flu-AmB demonstrated an effective reduction in fungal colonization after 17 days. This study demonstrates that Fluconazole-resistant Candida albicans strains are more pathogenic than susceptible ones in mice. Combination therapies with Fluconazole-Amphotericin B were most effective in reducing fungal colonization, outperforming monotherapy. These results support combination treatment for resistant Candida infections.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13631167/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148410220","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":"Preparation, characterization and application of feline CD3-specific single-domain antibodies.","authors":"Jingjing Wang, Qihuan Zhao, Bo Wang, Chengquan Du, Hui Wang, Jingsi Mei, Ruifeng Gao, Fuxiang Bao","doi":"10.1186/s13568-026-02093-7","DOIUrl":"10.1186/s13568-026-02093-7","url":null,"abstract":"<p><p>Feline lymphoma, a prevalent feline malignancy, responds poorly to conventional chemotherapy-a setting that underscores the urgent need for targeted therapeutic alternatives. The stable expression of CD3 on T-cell lymphoma cells positions this molecule as an ideal immunotherapeutic target. Compared with conventional antibodies, single-domain nanobodies derived from camelid heavy-chain antibodies offer distinct advantages, including smaller size, enhanced tissue penetration, and superior stability, thereby rendering CD3-targeted nanobodies particularly promising for managing feline T-cell lymphoma. We immunized a Bactrian camel with recombinant feline CD3ε extracellular domain and constructed a phage display nanobody library (capacity 5.97 × 10⁸). Sequencing of the constructed library revealed high diversity; among 10 randomly selected clones, 7 unique sequences were identified, and 93.33% of the clones carried intact VHH inserts. After three panning rounds, three high-affinity single-domain antibodies were selected and expressed, with clone 73 retaining stable binding at 156 ng/mL concentration. Flow cytometry showed that clone 73 specifically recognized approximately 37.95% of feline peripheral blood CD3⁺ cells. Western blotting detected a specific ~ 20 kDa band in feline lymphocyte lysates, and immunofluorescence confirmed precise cellsurface localization without non-specific signal. These findings demonstrate successful generation of feline CD3-specific nanobodies, which enable flow cytometric diagnosis of feline T-cell lymphoma and provide a foundation for developing immunotherapeutics.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13582682/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148403259","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}