Analytical biochemistry最新文献

{"title":"Enhanced performance of a plate-based TEV protease assay using linker-optimized, site-specifically biotinylated NanoLuc substrates.","authors":"Mitsuki Nakamura, Yoshiki Miya, Masafumi Sakono","doi":"10.1016/j.ab.2026.116279","DOIUrl":"https://doi.org/10.1016/j.ab.2026.116279","url":null,"abstract":"<p><p>We developed a plate-based luminescence assay for TEV protease using NanoLuc substrates site-specifically biotinylated through BirA co-expression in Escherichia coli. Substrates containing a TEV recognition sequence and AviTag, with or without a G4S linker, were immobilized on streptavidin-coated plates. Protease activity was quantified from the luminescence of NanoLuc released after cleavage. G4S insertion improved biotinylation efficiency and the luminescence response of immobilized substrates. Four-parameter logistic analysis showed lower EC20 and EC50 values and improved limits of detection and quantification. These findings demonstrate that linker insertion enhances substrate preparation and analytical sensitivity in this assay.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"116279"},"PeriodicalIF":3.1,"publicationDate":"2026-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149219808","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":"Corrigendum to \"Validation of SARS-CoV-2 neutralization assay using VSV-based pseudovirus system\" [Anal. Biochem. (2026), 719, 116239].","authors":"Anita Artarini, Marselina Irasonia Tan, Ernawati Arifin Giri-Rachman, Dessy Natalia, David Virya Chen, Chikako Ono, Yoshiharu Matsuura","doi":"10.1016/j.ab.2026.116270","DOIUrl":"https://doi.org/10.1016/j.ab.2026.116270","url":null,"abstract":"","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"116270"},"PeriodicalIF":3.1,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149164667","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":"Expression of concern: \"Specific detection of Mycobacterium sp. genomic DNA using dual labeled gold nanoparticle based electrochemical biosensor\" [Anal. Biochem. 417 (1) (2011) 73-79].","authors":"","doi":"10.1016/j.ab.2026.116186","DOIUrl":"https://doi.org/10.1016/j.ab.2026.116186","url":null,"abstract":"","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"717 ","pages":"116186"},"PeriodicalIF":3.1,"publicationDate":"2026-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148403295","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":"Corrigendum to \"One-step purification of a bioactive PAK1-derived peptide\" [Anal. Biochem. (2026), 709, 116014].","authors":"Djamali Muhoza, Emily P Esquivel, Stacy R Hunter, Patience S Okoto, Thallapuranam K S Kumar, Paul D Adams","doi":"10.1016/j.ab.2026.116258","DOIUrl":"https://doi.org/10.1016/j.ab.2026.116258","url":null,"abstract":"","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"116258"},"PeriodicalIF":3.1,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149057066","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":"Erratum to \"Development and malidation of a mechanism-based Ba/F3-hGHR bioassay for the potency assessment of recombinant human growth hormone\" [Anal. Biochem. 720 116237].","authors":"Tao Luo, Chen Guo, Xue Feng, Jianguang Lu, Yuanzhen Dong, Jun Feng","doi":"10.1016/j.ab.2026.116257","DOIUrl":"https://doi.org/10.1016/j.ab.2026.116257","url":null,"abstract":"","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"116257"},"PeriodicalIF":3.1,"publicationDate":"2026-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148982165","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 indole-oxadiazole-derived fluorescent probe for the detection of the human serum albumin level in thoracic surgery","authors":"Hanzhang Sun ,&nbsp;Guanglei Zuo ,&nbsp;Yueming Wu ,&nbsp;Yangyong Lou ,&nbsp;Jiang Feng ,&nbsp;Yulong Zheng","doi":"10.1016/j.ab.2026.116090","DOIUrl":"10.1016/j.ab.2026.116090","url":null,"abstract":"<div><div>In this work, the fluorescent probe <strong>InOx-HSA</strong> was developed from an indole-oxadiazole derivative for the detection of the HSA level in thoracic surgery. The probe reported a ratio fluorescence response from 405 nm to 505 nm under the excitation at 310 nm. <strong>InOx-HSA</strong> occupied the binding site of phenylbutazone in HSA and generated the fluorescence enhancement via Förster resonance energy transfer/twisted intramolecular charge transfer mechanisms. The response between <strong>InOx-HSA</strong> and HSA completed within 30 min. The probe-HSA complex kept stable within the range of 7.0-9.0. The ratio response was in a dosage-dependent manner with the limit of detection as 0.0002 mg/mL. <strong>InOx-HSA</strong> exhibited high specificity and low cytotoxicity. The site-binding strategy was confirmed by the drug competition, trypsin digestion, and protein denaturation by high temperature. <strong>InOx-HSA</strong> achieved visualizing HSA in living pulmonary cells, and revealed the ventilation-related consumption of HSA during the thoracic surgery.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"713 ","pages":"Article 116090"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147282090","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":"Imidazole stabilizes NAD(P)H and improves reductase assays under challenging conditions","authors":"Shengkai Yin ,&nbsp;Litao Wang ,&nbsp;Xian'ai Shi ,&nbsp;Guozeng Wang","doi":"10.1016/j.ab.2026.116094","DOIUrl":"10.1016/j.ab.2026.116094","url":null,"abstract":"<div><div>The reduced form of nicotinamide adenine dinucleotide (NAD(P)H) is prone to hydrolysis and ring-opening degradation under high-temperature and acidic conditions, resulting in a loss of reducing power and substantial inaccuracies in enzymatic assays. In this study, we systematically investigated the stabilizing effect of imidazole on NADH and NADPH under elevated temperature (30–60 °C) and acidic (pH 3–5) conditions, and further evaluated its applicability in reductase activity assays. Incubation experiments with varying imidazole concentrations (0–300 mM) demonstrated a pronounced concentration-dependent stabilization of NAD(P)H. Enzymatic assays revealed that imidazole enhanced the apparent activities of lactate dehydrogenase (LDH) and imine reductase (IRED) under both acidic and high-temperature conditions. Moreover, imidazole addition broadened the optimal pH and temperature range of these enzymes, markedly improving the accuracy and reproducibility of activity measurements. Collectively, these findings identify imidazole as an efficient stabilizing additive for NAD(P)H, enabling more reliable assessment of NAD(P)H-dependent reductase activities under high-temperature or acidic conditions.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"713 ","pages":"Article 116094"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147324467","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":"Physiological role of gamma-glutamyl transpeptidase: Demise of the gamma-glutamyl cycle","authors":"Marie H. Hanigan","doi":"10.1016/j.ab.2026.116087","DOIUrl":"10.1016/j.ab.2026.116087","url":null,"abstract":"<div><div>Gamma-glutamyl transpeptidase (GGT) is an enzyme with a plethora of names and purported functions. The enzyme has long been identified with an amino acid transport system called the gamma-glutamyl cycle which was proposed in the 1970s. The foundation of this cycle held that GGT catalyzed a transpeptidation reaction and in the process transported an extracellular amino acid into the cell as a gamma-glutamyl amino acid. However, subsequent studies showed that under physiological conditions the enzyme is a hydrolase. It cleaves the gamma-glutamyl bond of glutathione, glutathione <em>S</em>-conjugates and other gamma-glutamyl compounds releasing glutamate. The release of glutamate from glutathione, renders the other product of the reaction, cysteinyl-glycine, susceptible to cleavage by cell surface dipeptidases. The development of the GGT-deficient mouse showed definitively that the primary role of GGT <em>in vivo</em> is cysteine homeostasis, not amino acid transport. GGT also plays a role in oxidative stress, drug metabolism, and inflammation. Despite numerous studies showing that the gamma-glutamyl cycle is incorrect, it nonetheless persists in the literature, in textbooks, and currently on the GGT Wikipedia page. Confusion regarding the physiological role of GGT can lead to misinterpretation of data and blunt progress in many important areas of investigation. This review focuses on the physiological functions of GGT and summarizes the data demonstrating the gamma-glutamyl cycle does not exist.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"713 ","pages":"Article 116087"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146200181","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":"A new paradigm in tuberculosis diagnostics: Biosensing advances for early detection of Mycobacterium tuberculosis","authors":"Pooja Khandelwal,&nbsp;Neelam Yadav,&nbsp;Arzoo Saini,&nbsp;Neelam Singh Sangwan","doi":"10.1016/j.ab.2026.116088","DOIUrl":"10.1016/j.ab.2026.116088","url":null,"abstract":"<div><div>Tuberculosis (TB) remains one of the most serious and deadly infectious diseases worldwide. Despite being preventable and curable, it is responsible for increasing the annual fatality by millions. The situation is even more alarming in the developing nations, where TB ranked in the top 10 in terms of global mortality. <em>Mycobacterium tuberculosis</em> is the causal pathogen for tuberculosis. It is an extremely resilient bacterium that eludes the immunity of the host and prevails in latent forms. Several biomarkers, including antigens (CFP-10, ESAT-6), antibodies, and nucleic acids, have been identified for their rapid and accurate detection. Conventional screening approaches like culture, chest X-ray, sputum smear microscopy, and PCR-based assays are widely used. However, these approaches suffer from various shortcomings, <em>viz.,</em> less sensitivity, delayed diagnosis, potential false negatives, requirement of biosafety infrastructure, and skilled personnel. These diagnostic challenges are resolved by biosensors as they are quick, sensitive, and resource-efficient alternatives to conventional methods. This review comprehensively discusses the pathogenicity of TB as well as conventional approaches for its diagnosis. Further, we have also provided an in-depth analysis of recently reported biosensors for TB detection and critically evaluated their analytical performance.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"713 ","pages":"Article 116088"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146218505","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":"Untargeted metabolomic and lipidomic profiling in a hyperuricemic rabbit model: a pilot study","authors":"Junyoung Ahn ,&nbsp;Bo Young Hwang ,&nbsp;Kyo Bin Kang ,&nbsp;Nak-Hoon Son ,&nbsp;Hyang-Ja Lee ,&nbsp;Jongwon Oh ,&nbsp;Mi-Young Kim ,&nbsp;Sanghoo Lee ,&nbsp;Mikyeong Lee ,&nbsp;Kyoung-Ryul Lee ,&nbsp;Sung Kweon Cho ,&nbsp;Jeong-Ho Kim","doi":"10.1016/j.ab.2026.116072","DOIUrl":"10.1016/j.ab.2026.116072","url":null,"abstract":"<div><div>The incidence of hyperuricemia is rising globally; however, its molecular mechanisms and reliable progression biomarkers remain unclear. This study characterized temporal changes in serum metabolites and lipids in hyperuricemic rabbit models using untargeted metabolomic and lipidomic profiling with ultra-high-performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF-MS/MS). Serum samples collected at baseline (0 h), 48 h, and 84 h (n = 3 per time point) were analyzed using both hydrophilic interaction chromatography (HILIC) and reversed-phase C18 columns to achieve broad metabolite and lipid coverage. HILIC analysis yielded 9813 peaks with 65 annotated compounds, whereas C18 analysis identified 7333 peaks with 122 annotated compounds across both positive and negative ion modes. Significant monotonic increases were observed in the normalized feature intensities of 4-aminohippuric acid, L-alanine, N-isobutyrylglycine, and uric acid, whereas the normalized feature intensity of inosine decreased relative to baseline. These metabolites satisfied two selection criteria, [|log₂(fold change, FC)| ≥ 0.58, Nemenyi <em>p</em> &lt; 0.05] and variable importance in projection (VIP) &gt; 1.0 using orthogonal partial least squares-discriminant analysis (OPLS-DA). Among them, inosine and 4-aminohippuric acid, which showed a strong correlations with uric acid, emerged as the most exploratory candidate biomarkers for hyperuricemia progression. The C18-based lipidomic profiling identified the phosphatidylinositols, PI(16:0_20:4) and PI(18:1_18:2), as the only lipid species that met the two selection criteria. In particular, PI(16:0_20:4) was strongly correlate with uric acid. Collectively, our study reveals the metabolomic and lipidomic changes for hyperuricemia and suggests exploratory candidate biomarkers for its early detection and monitoring.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"713 ","pages":"Article 116072"},"PeriodicalIF":2.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146123408","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}