Analytical biochemistry最新文献

{"title":"A recombinant full-length VP2-2b-based ELISA for evaluating immunoprotection against canine parvovirus-2: Expression, development and validation","authors":"Ezgi Salmanli ,&nbsp;Taner Karaoglu","doi":"10.1016/j.ab.2026.116070","DOIUrl":"10.1016/j.ab.2026.116070","url":null,"abstract":"<div><div>Canine parvovirus-2 remains a major threat to canine health, with maternal antibody interference and limitations of traditional diagnostic methods posing challenges to effective disease control. Maternal-derived antibodies can persist at levels that interfere with the development of active immunity following vaccination. Therefore, accurate detection of existing antibody levels prior to vaccination is essential to ensure induction of protective immunity. Hemagglutination inhibition tests are labor-intensive and highly susceptible to external variability, emphasizing the need for reliable and cost-effective alternatives. In this study, we developed and optimized an indirect ELISA using a full-length, recombinant VP2-2b protein expressed in <em>E. coli</em> to detect CPV-2 antibodies. To our knowledge, this is the first ELISA developed around a soluble full-length CPV-2b protein, with comprehensive optimization and clinical validation. VP2-2b was selected for its unique bidirectional neutralization kinetics, enhancing the diagnostic accuracy of CPV-2 serology. The ELISA demonstrated excellent sensitivity and specificity, effectively distinguishing between seropositive and seronegative samples. A gray zone (OD450 = 0.26–0.30) was defined to represent borderline antibody titers, where retesting is recommended to determine accurate vaccination timing. Optimal assay conditions were established as 7.5 μg antigen coating concentration and 1:300 serum dilution, yielding a high signal-to-noise ratio (SNR) of 5.6. Validation against HI tests showed strong correlation with minimal non-specific binding. This ELISA system provides a reliable and economical alternative to traditional methods, helping overcome the limitations of the HI test in field settings and supporting informed decisions in vaccination planning.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116070"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146123466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an AI image Recognition–based lateral flow immunochromatographic test strip for higenamine detection","authors":"Rui Zhang ,&nbsp;Yiyang Tong ,&nbsp;Yumeng Shi ,&nbsp;Yinuo Cai ,&nbsp;Qi Zhao ,&nbsp;Yang Lu","doi":"10.1016/j.ab.2026.116069","DOIUrl":"10.1016/j.ab.2026.116069","url":null,"abstract":"<div><div>Higenamine (HM) is a naturally occurring benzylisoquinoline alkaloid found in plants and is classified as an S3 prohibited substance in the 2020 World Anti-Doping Agency report. To prevent doping violations in competitive sports, it is essential to develop timely and accurate detection methods. In this study, we propose, for the first time, a detection method that combines artificial intelligence (AI)-based image recognition technology with lateral flow immunoassay (LFIA). This method utilizes gold nanoparticles (AuNPs) conjugated with HM-specific antibodies for the rapid detection of HM in urine via LFIA. It integrates AI-based image recognition to quantitatively analyze color intensity of the test strip's detection line. Experimental results demonstrated that the proposed method achieves a limit of detection of 0.49 ng/mL, enabling accurate identification within the concentration range of 2–8 ng/mL. For model training, an Image Classification of HM Test strip dataset was created. To address the limitations of this dataset, namely its small size (n = 304) and limited feature diversity, an advanced solution tailored to these constraints was proposed. The use of Contrast Limited Adaptive Histogram Equalization (CLAHE), which enables finer-grained feature extraction, ensured excellent recognition accuracy of the model in this dataset. During actual urine detection, the model achieved a prediction accuracy of 96.88% on the test set. This approach provides an efficient and reliable technical solution for on-site rapid detection of HM, addressing the critical need for timely monitoring in anti-doping efforts.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116069"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146140947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of LMW chitosan-stabilized gold nanoparticle DNA biosensors functionalized with thiolated probe for Vibrio parahaemolyticus detection","authors":"Jessa D. Alonzo ,&nbsp;Kristopher Ray S. Pamintuan ,&nbsp;Khyle Glainmer N. Quiton ,&nbsp;Jyh Jian Chen","doi":"10.1016/j.ab.2026.116064","DOIUrl":"10.1016/j.ab.2026.116064","url":null,"abstract":"<div><div>Acute hepatopancreatic necrosis disease (AHPND), caused by <em>Vibrio parahaemolyticus</em> (Vp) carrying <em>pir</em>^<em>vp</em><em>A/pir</em>^<em>vp</em><em>B</em>, continues to threaten shrimp aquaculture across Asia. This study demonstrates a laboratory proof-of-concept gold nanoparticle-based (AuNPs) colorimetric biosensor targeting the <em>pir</em>^<em>vp</em><em>A</em> gene of Vp. Citrate-reduced AuNPs (∼15.54 nm; SPR ≈ 520 nm) were functionalized with thiolated ssDNA probes (AuNP-Probe) using a deoxyadenosine triphosphate-assisted immobilization strategy and subsequently stabilized with low molecular weight (LMW) chitosan (AuNP-Probe-Chit). Loop-mediated isothermal amplification (LAMP) generated the target DNA, while detection relied on salt-induced aggregation assessed by visual color/precipitate formation and UV–Vis spectral shifts (A₆₅0/A_λmax, <em>λ</em>_max ≈ 532 nm for AuNP-Probe-Chit). Optimization identified 1.0 μM probe and 0.01% (v/v) chitosan as the optimal conditions for forming stable conjugates. Upon hybridization, an 8:2 (v/v) AuNP-Probe-Chit:LAMP mixture, combined with 0.5 M NaCl, enabled clear discrimination between positive and negative samples. Positives retained a pink-red supernatant with minimal aggregation, whereas negatives formed precipitates and exhibited higher aggregation indices. The biosensor demonstrated strong specificity for <em>pir</em>^<em>vp</em><em>A</em>-positive DNA, without cross-reacting with <em>Vibrio vulnificus</em> or <em>Vibrio alginolyticus</em>, and exhibited sensitivity approaching the LAMP amplification threshold, with reliable discrimination observed at ≥10⁻¹ ng of target DNA. These findings establish a robust, equipment-lean diagnostic platform that integrates sequence-specific hybridization with polymer-mediated colloidal stabilization. The AuNP-Probe-Chit system offers rapid, visible discrimination, highlighting its potential for future AHPND surveillance in resource-limited settings. Compared with previous works, the incorporation of thiolated ssDNA probe functionalization with chitosan stabilization within a single AuNPs-based platform tailored for aquaculture pathogens, such as Vp, is currently underexplored, highlighting a significant need that this study aims to address.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116064"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146099535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design and in vitro evaluation of an Fc-fusion epitope-based vaccine candidate against Helicobacter pylori","authors":"Roghayeh Mohammadzadeh ,&nbsp;Shaho Menbari ,&nbsp;Janbibi Dorazehi ,&nbsp;Mojtaba Sankian ,&nbsp;Hadi Farsiani","doi":"10.1016/j.ab.2026.116071","DOIUrl":"10.1016/j.ab.2026.116071","url":null,"abstract":"<div><div><em>Helicobacter pylori</em>, a Group I carcinogen that infects over 50% of the global population, is a major contributor to gastric cancer and the growing crisis of antibiotic resistance. However, vaccine development has remained challenging due to the poor immunogenicity of candidate antigens and limitations in delivery strategies. To address this, we engineered a novel Fc-fusion vaccine targeting antigen-presenting cells (APCs) by fusing <em>H. pylori</em> UreB₃₁₇-₃₂₉ and UreB₄₀₉-₄₂₁ epitopes with Lpp20 lipoprotein to the murine IgG2a Fc domain (UreB: Lpp20: mFcγ2a). This study presents a preliminary, proof-of-concept evaluation of an Fc-fusion epitope-based vaccine candidate, focusing on <em>in vitro</em> characterization and APC-targeting properties. The construct was codon-optimized, expressed in <em>Pichia pastoris</em>, and purified via affinity chromatography. Expression levels in the mg/mL range, as determined by biochemical analysis, with &gt;95% purity confirmed by SDS-PAGE and Western blot. Co-localization assays demonstrated 4.2-fold higher APC uptake of the Fc-fusion protein versus a His-tagged control (<em>p</em> &lt; 0.001, unpaired <em>t</em>-test), mediated by FcγRI (CD64) binding. The protein maintained its structural stability for 30 days at 4 °C (&lt;5% batch variability), underscoring manufacturing feasibility. While <em>in vitro</em> results validate APC targeting, limitations include preclinical scope and murine Fc compatibility gaps for human translation. This study established <em>P. pastoris</em> as a scalable platform for Fc-fusion vaccines, bypassing adjuvants and enhancing immunogenicity through conserved epitopes. Future work must evaluate <em>in vivo</em> efficacy in infection models and human Fc adaptations. These findings offer a blueprint for next-generation vaccines against <em>H. pylori</em> and other intracellular pathogens, urgently needed in antimicrobial resistance containment strategies.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116071"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146117643","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aging differentially affects the uptake of methionine by select rat tissues","authors":"Juan A. Azcona,&nbsp;Anja S. Wacker,&nbsp;Thomas M. Jeitner,&nbsp;James M. Kelly","doi":"10.1016/j.ab.2026.116062","DOIUrl":"10.1016/j.ab.2026.116062","url":null,"abstract":"<div><div>Experimental and epidemiological studies indicate that significant reductions in dietary methionine intake profoundly impact both health and life spans. Methionine levels also decrease in the blood of mammals as they age. Here, we report that methionine uptake by four of the major methionine-consuming organs, the liver, kidneys, heart, and brain, is increased in old rats as compared to young adult rats, as measured by [¹¹C]methionine uptake by these organs using positron emission tomography. This increased uptake was sustained for at least 30 min and resulted in 1.6- to 1.9-fold increases in methionine uptake by the major methionine-consuming organs in old rats, suggesting an age-associated acceleration of methionine metabolism and its associated biochemical pathways. By contrast, the uptake of [¹⁸F]fluorodeoxyglucose by the liver, kidneys, heart, and brain of older rats was reduced relative to that by their younger adult counterparts, indicating a decline in glucose metabolic activity with age. Remarkably, the uptake of 2-[¹⁸F]fluoropropionic acid by these organs remained essentially unchanged between young and aged adult rats, suggesting the presence of a stable biochemical equilibrium in propionate metabolism across the aging spectrum. These findings underscore the utility of positron emission tomography in revealing significant age-related alterations in organ-specific biochemical processes.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116062"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146040338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An amplification-free and in situ detection system for miRNA-451 with single-nanoparticle LSPR biosensing on a multi-channel microfluidic chip","authors":"Kuilin Liu ,&nbsp;Xingyu Zi ,&nbsp;Jianqi Wang ,&nbsp;Zhijia Bao ,&nbsp;Haiying Chen ,&nbsp;Xinyu Gao ,&nbsp;Guohua Liu ,&nbsp;Mingqing Zhang","doi":"10.1016/j.ab.2026.116075","DOIUrl":"10.1016/j.ab.2026.116075","url":null,"abstract":"<div><div>Early screening for colorectal cancer (CRC) is crucial for reducing its mortality. This study developed an amplification-free, highly sensitive sensing system based on localized surface plasmon resonance (LSPR) technology integrated with a multi-channel microfluidic chip platform for the in situ detection of the key CRC biomarker microRNA-451 (miRNA-451) using gold nanoparticles (AuNPs), achieving a limit of detection (LOD) of 19.2 fM. The core of this system is a multi-channel microfluidic chip serving as the detection substrate, where single-stranded DNA (ssDNA) probes are immobilized on the AuNPs surfaces to construct the sensor. In the presence of the target miRNA-451, it hybridizes specifically with two types of ssDNA probes, inducing the formation of AuNPs dimers and consequently causing a red shift in their LSPR scattering spectra. Unlike conventional methods, this study employed a precise single-nanoparticle tracking strategy, quantifying the target by statistically analyzing the proportion of AuNPs exhibiting significant spectral red shifts within the same field of view (FOV). This approach effectively mitigates spatial sampling errors arising from FOV inconsistencies. Furthermore, the system decouples the “hybridization\" and “detection\" functions, significantly enhancing detection efficiency and accuracy. This work provides a robust technical platform for the amplification-free and highly sensitive detection of miRNA-451.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116075"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146140968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Versatile applications of Light-Oxygen-Voltage (LOV) domain proteins in optical microscopy","authors":"Ayushi Mishra ,&nbsp;Suneel Kateriya","doi":"10.1016/j.ab.2026.116065","DOIUrl":"10.1016/j.ab.2026.116065","url":null,"abstract":"<div><div>Various blue-light photoreceptor proteins have photo-responsive domains known as light, oxygen, voltage (LOV) domains, which are extensively distributed in plants, algae, fungi, and bacteria. When exposed to blue light, the flavin chromophore and a highly conserved cysteine residue form a covalent adduct on a microsecond time scale. LOV domains are common photosensory modules that can be applied to optogenetics, regulated synthesis of reactive oxygen species, and fluorescence microscopy. This review explores the photocycle kinetics and applications of various LOV domains, which have been explored for confocal microscopy, two-photon microscopy, and super-resolution microscopy. Many LOV domains have been derived and modulated for use in different types of microscopic applications. Molecular understanding, diversity of LOV domains, and versatile photo-physical characteristics of these proteins have immense potential for the development of useful probes for various microscopy tools. There is a great demand for perspective research on LOV domain proteins for harnessing their possible optobiotechnological applications.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116065"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146075775","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Species-specific isothermal nucleic acid amplification assay targeting Internal Transcribed Spacer (ITS) for rapid authentication of the medicinal crop Cirsium japonicum and Cirsium setosum in herbal markets","authors":"Jia-An Ling ,&nbsp;Jin-Xuan He ,&nbsp;Chia-Hsin Lin ,&nbsp;Shyang-Chwen Sheu ,&nbsp;Jai-Hong Cheng ,&nbsp;Meng-Shiou Lee","doi":"10.1016/j.ab.2026.116063","DOIUrl":"10.1016/j.ab.2026.116063","url":null,"abstract":"<div><h3>Background</h3><div><em>Cirsium japonicum</em> (CJ) and <em>Cirsium setosum</em> (CS) are two widely recognized medicinal crops in Chinese herbal medicine (CHM) that are listed as authentic herbal plants in the herbal pharmacopeia. Given their strikingly similar morphological characteristics, CJ and CS are particularly susceptible to adulteration in the herbal marketplace, raising significant concerns about product integrity and consumer safety.</div></div><div><h3>Purpose</h3><div>To ensure the safety and efficacy of CHM, proper authentication of these herbs is a critical step in maintaining high-quality pharmaceutical production.</div></div><div><h3>Methods</h3><div>In this study, we developed a species-specific loop-mediated isothermal amplification (LAMP) method for the authentication of CJ and CS.</div></div><div><h3>Results</h3><div>We designed specific LAMP primer sets for both species using the selected DNA barcode, the internal transcribed spacer (ITS) sequence, through <em>in silico</em> analysis. After validating the specificity of the primers, we successfully authenticated CS and CJ using the LAMP primer sets and visualized the detection results through gel electrophoresis. The sensitivity of the LAMP method for CJ and CS authentication was established, with a limit of detection (LOD) of 100 pg for CJ and 50 pg for CS, using direct SYBR Green staining. When we applied the LAMP method to commercial <em>Cirsium</em> products, we found that only 25% (5 out of 20) of the samples contained CJ, while none contained CS.</div></div><div><h3>Conclusion</h3><div>In conclusion, we successfully established a species-specific LAMP assay for authenticating CJ and CS. This method can be used for species verification and is applicable to commercial <em>Cirsium</em> products for authenticity testing, contributing to quality control in pharmaceutical manufacturing.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116063"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146099514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dual-strategy qMSP reference optimization for Low-DNA clinical specimens","authors":"Shan Zhang ,&nbsp;Yixuan Tang ,&nbsp;Jiahui Jiang ,&nbsp;Ling Chen","doi":"10.1016/j.ab.2026.116068","DOIUrl":"10.1016/j.ab.2026.116068","url":null,"abstract":"<div><div>Detecting tumor DNA methylation in low DNA concentration samples at high qualification rates is a significant challenge in translating and conducting regulatory trials for tumor methylation detection kits. Existing internal reference systems frequently malfunction, resulting in the mislabelling of valuable clinical specimens as unusable. We developed a two-part framework for improving reference primer-probe sets, by combining a systematic improvement of existing <em>ACTB</em> assays with new primer designs that target previously unexplored, CpG-free genomic areas. A three-stage screening process involving SYBR Green qPCR, TaqMan probe-based qPCR, and validation across different biological samples led to the identification of two superior primer sets: an optimized combination (bisACTB_103_F2+R2+P2) and a novel set (bisProAB1_F2+R2+P1). Exhibited enhanced amplification efficiency and specificity in cell lines, tissues, blood cells, and plasma were observed in both. Notably, bisProAB1_F2+R2+P1 considerably enhanced amplification efficiency in low-concentration plasma cell-free DNA, resulting in a nearly 5 % increase in the detection success rate within a large clinical study group and allowing for reliable analysis of minimal-input samples. This work offers verified, high-performance reference systems to boost the reliability of DNA methylation-based diagnostics and make the most of precious biospecimens.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116068"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146099521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reference Plasma Proteome Map of Crotalus durissus terrificus","authors":"Isabela Tokuhara Makida ,&nbsp;Amanda Almeida Resende ,&nbsp;Letícia Gomes de Pontes ,&nbsp;Laudicéia Alves de Oliveira ,&nbsp;Leonardo Melo ,&nbsp;Francilene Capel Tavares de Carvalho ,&nbsp;Ivânio Teixeira Borba-Junior ,&nbsp;Regina Kiomi Takahira ,&nbsp;Rui Seabra Ferreira Junior ,&nbsp;Lucilene Delazari dos Santos","doi":"10.1016/j.ab.2026.116051","DOIUrl":"10.1016/j.ab.2026.116051","url":null,"abstract":"<div><div>Snakebite envenoming remains a major public health concern worldwide, while the maintenance of captive snakes is essential for conservation, venom production, and biomedical research. However, little is known about the plasma proteome of snakes, a gap that limits physiological assessments and the development of species-specific reference protein profile. This study aimed to characterize the plasma proteome of healthy adult <em>Crotalus durissus terrificus</em> snakes to establish a molecular baseline that supports biological evaluation and future diagnostic research. Blood samples were collected from 19 specimens kept in capitivity, followed by hematological, biochemical, and proteomic analyses. Hematology confirmed preserved cell morphology and values within established reptilian reference ranges, indicating good health status. Biochemical analyses showed albumin, total protein, and aspartate aminotransferase levels consistent with previous reports, while uric acid values were comparatively lower, reflecting reptilian renal physiology. Proteomic profiling using 2D electrophoresis and LC-MS/MS identified 301 protein spots, of which 273 were successfully annotated, covering approximately 90% of the detectable plasma proteome. Functional classification revealed proteins associated with immune function, transport, metabolism, and anti-hemorrhagic activity. The plasma proteomic baseline generated here contributes to the understanding of snake systemic physiology and provides foundational data to support future diagnostic, veterinary and biotechnological investigations, with direct benefits for animal welfare, biosafety, and reproducibility in venom-based biotechnological applications.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"712 ","pages":"Article 116051"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145987600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}