Mohd Imran Shah, Kavitha Ramaswamy, Sangita Venkataraman, Sankaranarayanan Meenakshisundaram
{"title":"Cloning, expression and molecular evolution of <i>Komagataella phaffii</i> ketoreductase with enhanced catalytic activity.","authors":"Mohd Imran Shah, Kavitha Ramaswamy, Sangita Venkataraman, Sankaranarayanan Meenakshisundaram","doi":"10.1080/10826068.2025.2525210","DOIUrl":"https://doi.org/10.1080/10826068.2025.2525210","url":null,"abstract":"<p><p>The synthesis of optically active alcohols is vital for the pharmaceutical and fine chemical industries, yet traditional chemical methods generally lack efficiency and stereoselectivity. Although ketoreductases are a more environmentally friendly option, many of them need to have their catalytic performance improved to satisfy industrial demands. This study focused on engineering <i>Komagataella phaffii</i> ketoreductase (KpKR), the first characterized ketoreductase from this yeast, by applying an innovative combination of error-prone PCR and ribosome display for rapid directed evolution. Through the use of kinetic analysis, molecular docking, homology modeling, and substrate specificity assays, five evolved variants (M1-M5) were identified and characterized. Compared to a range of microbial ketoreductases, such as Bacillus sp. ECU0013 and yeast-derived enzymes, the KpKR variants demonstrated efficient catalytic performance; notably, M5 showed enhanced catalytic efficiency (k<sub>cat</sub>/k<sub>m</sub> = 199.58 s<sup>-1</sup>mM<sup>-1</sup>) toward ketoesters, while M1 demonstrated remarkable activity for halogenated substrates, outperforming Bacillus enzymes by thousands-fold. Structural investigation identified key C-terminal changes that likely contributed to improved active site accessibility. Furthermore, sequence analysis confirmed KpKR as a member of the short-chain dehydrogenase/reductase (SDR) family. These results underscore the innovative utility of ribosome display in enzyme engineering and establish the evolved KpKR variants as powerful, highly efficient biocatalysts suitable for challenging pharmaceutical synthesis.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-14"},"PeriodicalIF":2.0,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144565103","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}
Nidhi Mallya, Stefanie Rudolph, Sowmya R Holla, Subbalaxmi Selvaraj
{"title":"Electrochemical system for detection of fermentation-derived bioethanol via cyclic voltammetry using nickel oxide modified glassy carbon electrode.","authors":"Nidhi Mallya, Stefanie Rudolph, Sowmya R Holla, Subbalaxmi Selvaraj","doi":"10.1080/10826068.2025.2525205","DOIUrl":"https://doi.org/10.1080/10826068.2025.2525205","url":null,"abstract":"<p><p>Accurate detection of bioethanol is essential for optimizing biofuel production. Although precise, current industrial methods such as Gas chromatography (GC) and High-performance liquid chromatography (HPLC) are expensive, time-consuming, and unsuitable for on-site monitoring. This study explores the development of a cost-effective, rapid electrochemical method using nickel oxide (NiO)-modified glassy carbon electrodes for detecting bioethanol produced via fermentation using <i>Saccharomyces cerevisiae</i>. Fermentation was optimized with varying sucrose concentrations (20-100 g/L), achieving a maximum ethanol yield of 15.69 g/L at 40 g/L sucrose with an agitation of 100 rpm at 48 h. Distilled ethanol was characterized by GC and HPLC. Cyclic voltammetry (CV) was employed for electrochemical detection, with NiO nanoparticles synthesized and characterized using X-ray diffraction (XRD), confirming their crystalline structure and electroactive surface area. The CV analysis demonstrated clear anodic and cathodic peaks for standard and fermented bioethanol, with a cathodic peak at 0.43 V and an anodic peak at 0.52 V. The oxidation peak current showed a linear relationship with the scan rate, confirming a diffusion-controlled process. These findings confirm NiO-modified electrodes as a dependable ethanol detection method. As a scalable and sustainable substitute for traditional methods, the technique exhibits potential for non-enzymatic bioethanol monitoring in industrial settings.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-14"},"PeriodicalIF":2.0,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144529406","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}
Ashitha Jose, Maya Mathew, Aswani R, Bipinbal Parambath Kanoth, Sebastian Ks, Radhakrishnan E K
{"title":"Zinc oxide nanoparticle mediated modulation of antimicrobial and physico-chemical properties of essential oil containing PVA nanocomposites.","authors":"Ashitha Jose, Maya Mathew, Aswani R, Bipinbal Parambath Kanoth, Sebastian Ks, Radhakrishnan E K","doi":"10.1080/10826068.2025.2522461","DOIUrl":"https://doi.org/10.1080/10826068.2025.2522461","url":null,"abstract":"<p><p>Microbial infestation related food loss poses a major threat to the global food sector. Both bacterial and fungal infestations play a crucial role in the food deterioration and various health issues. This has led to the increased demand for the development of active packaging materials. In the current study, selected essential oils were subjected to MIC, MBC and MFC analysis against both bacterial and fungal agents. Further to this, PVA based bionanocomposites were developed by incorporating the selected essential oils along with ZnONPs. By the FTIR analysis, the interactions among individual components of the developed thin films could be confirmed. Interestingly, the incorporation of nanoparticles was found to have modulatory effect on the release of essential oil components from the films as evidenced by the GC-MS analysis. Further to this, the films were also demonstrated to have enhanced mechanical properties and low moisture content which favor its application as promising packaging material. The incorporation of ZnONPs was also observed to positively modulate the antibacterial and antifungal activity of the films without compromising its microbial barrier efficacy. The findings of the current study thus indicate the potential of the developed bionanocomposite films as antimicrobial packaging systems.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-20"},"PeriodicalIF":2.0,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144529407","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":"Evaluation of brilliant green decolorization by LiP purified from a novel <i>Penicillium chrysogenum</i> isolated from Similipal forest soil, Odisha.","authors":"Subhashree Rath, Hrudayanath Thatoi","doi":"10.1080/10826068.2025.2515949","DOIUrl":"https://doi.org/10.1080/10826068.2025.2515949","url":null,"abstract":"<p><p>Lignin peroxidase (LiP) is a haem-containing microbial ligninolytic enzyme that has gained much attention for its potential application to degrade industrial effluents. In the current study, seven fungi (SLF1-7) showing ligninolytic activity were isolated from Similipal forest soil, and screened for LiP activity. SLF3 showed the highest LiP activity (3.351 U/mL.min) at un-optimized conditions and was identified as <i>P. chrysogenum</i> using 18S rRNA sequencing. The production of enzyme was optimized using Response Surface Methodology. LiP activity of <i>P. chrysogenum</i> was enhanced to 9.38 U.mL<sup>-1</sup>.min (2.79 folds) at pH 6, temperature 37.5 °C, substrate concentration 1.75%, and time 108 h. The LiP enzyme was purified by ammonium sulfate precipitation followed by dialysis and Sephadex G-100 column chromatography. The purified protein showed a single band of approximately 40 kDa by SDS-PAGE analysis. The purified LiP also showed the highest catalytic efficiency with K<sub>m</sub> of 0.316 M and V<sub>max</sub> of 147.059 U/mL.min. The purified LiP reduced brilliant green color (69.53 ± 0.003%) after 80 min of incubation at 37 °C and pH 6.0. It is apparent from the study that the strain <i>P. chrysogenum</i> has significant potential in degrading brilliant green, which can be explored for bioremediation applications of industrial dyes.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-13"},"PeriodicalIF":2.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144317769","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}
Mohammad Harun Ur Rashid, Chiu Hock Ing, Nor Datiakma Mat Amin, Mohd Nazri Ismail
{"title":"Optimizing the extraction of high-potency antioxidants from <i>Solanum lycopersicum L.</i> fruit: a comparative study of microwave-assisted, ultrasound-assisted and accelerated solvent extractions.","authors":"Mohammad Harun Ur Rashid, Chiu Hock Ing, Nor Datiakma Mat Amin, Mohd Nazri Ismail","doi":"10.1080/10826068.2025.2518250","DOIUrl":"https://doi.org/10.1080/10826068.2025.2518250","url":null,"abstract":"<p><p>Tomatoes, rich in phenolics and antioxidants, offer a rich source of dietary antioxidants with potential health-promoting properties. Owing to the involvement of toxic solvents, high energy consumption, and hazardous waste, a safer and more sustainable extraction method has been preferred in the food and pharmaceutical industries. This study systematically investigated three advanced extraction techniques-microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), and accelerated solvent extraction (ASE)-to maximize phenolic yield and antioxidant capacity while concurrently minimizing the use of hazardous solvents, focusing Baccarat tomato variant. Among the tested methods, ASE demonstrated superior DPPH radical scavenging activity (87.480 ± 0.856%) with an IC50 of 4.65 mg/mL, while UAE achieved the highest total phenolic content (TPC, 9.060 ± 0.028 mg GAE/g DW). A strong correlation between TPC and antioxidant activities (DPPH and FRAP) was observed, emphasizing interdependence between phenolic content and bioactivity. Hierarchical cluster analysis (HCA) revealed 66.66% of ASE samples grouped into a cluster based on enhanced antioxidant properties. While UAE is superior for maximizing phenolic yield, ASE is the optimal method for producing extracts with the highest antioxidant potency, highlighting the critical role of technology selection in developing targeted ingredients for functional foods and therapeutic agents toward good health and well-being.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-10"},"PeriodicalIF":2.0,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144294811","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}
Pelin Koseoglu-Yılmaz, Mehmet Veysi Caglayan, Abdulselam Ertas, Ufuk Kolak
{"title":"Response surface methodology for extraction of chia seed phenolics and their quantification by LC-MS/MS.","authors":"Pelin Koseoglu-Yılmaz, Mehmet Veysi Caglayan, Abdulselam Ertas, Ufuk Kolak","doi":"10.1080/10826068.2025.2517275","DOIUrl":"https://doi.org/10.1080/10826068.2025.2517275","url":null,"abstract":"<p><p><i>Salvia hispanica</i> L. species, also known as \"Chia,\" is an annual plant belonging to the Lamiaceae family. Chia seeds are of great importance in the daily diet due to their high quality components, such as proteins, carbohydrates, fibers, minerals, antioxidants and fats. In the present study, optimum maceration conditions were determined for preparation of chia seed extracts with high total phenolic compound content by experimental design approach. For this purpose, a response surface methodology type known as Box-Behnken design was applied. Ethanol concentration of the extraction solvent, extraction time and solvent volume were examined as the independent variables, whereas the total phenolic content was the response. The developed experimental model was evaluated statistically and found to fit well, considering the lack of fit <i>p</i>-value of 0.184 and coefficient of determination as 0.9731 at the 95% confidence level. The extract prepared by optimized conditions as 56.1 minutes of extraction time and 49.5 mL of solvent volume containing 61.4% ethanol had a total phenolic content of 53.5 ± 1.9 μg of pyrocatechol equivalent/mg extract. The phenolic compounds of the prepared extract were quantified by liquid chromatography-tandem mass spectrometry. Rosmarinic acid was found to be the major component with a concentration of 22.702 mg/g extract.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-6"},"PeriodicalIF":2.0,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144267173","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":"Extraction kinetics and thermodynamic evaluation of functional components in thermosonicated Sohshang juice optimized by ANN-GA.","authors":"Puja Das, Prakash Kumar Nayak, Radha Krishnan Kesavan","doi":"10.1080/10826068.2025.2515946","DOIUrl":"https://doi.org/10.1080/10826068.2025.2515946","url":null,"abstract":"<p><p>This study investigates the underlying extraction kinetics and thermodynamic principles driving the process, building on earlier research that enhanced thermosonication (TS) settings to boost the bioactive components of Sohshang (<i>Elaeagnus latifolia</i>) fruit juice. Responses were assessed over a temperature range of 30 °C to 50 °C after the ideal thermosonicated sohshang fruit juice (TSSJ) processing parameters (50% amplitude, 40 °C, and 60 minutes) were determined using the integrated ANN-GA model. The study uses kinetic modeling to determine the best mathematical fit by analyzing the release patterns of functional components like ascorbic acid (AA), total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity (AOA). With the highest R<sup>2</sup> (0.99) and lowest χ<sup>2</sup> (0.64) values for each of the five responses of various kinetic modeling, the pseudo second-order model was determined to be the best appropriate model. To comprehend the energy requirements and viability of thermosonication-assisted extraction, important thermodynamic parameters were also measured, such as activation energy (Ea: 23.29 to 29.23), enthalpy (ΔH: 15.91 to 58.05 kJ/mol), entropy (ΔS: 42.87 to 112.24 J/mol K), and Gibbs free energy (ΔG: -12.97 to -40.00 kJ/mol).</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-13"},"PeriodicalIF":2.0,"publicationDate":"2025-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144249342","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":"Sequential extraction, structural characterization, and biological activities of polysaccharides from olive (<i>Olea europaea</i> L.) pomace.","authors":"Xiao-Liang Zhao, Kai-Li Qi, Hui Han, Hai-Dong Wen, Yan-Mei Liu, Qing-Song Zhou, Yuan-Yuan Ma, Cun-Jin Wang, Jing Zhang, Ji Zhang, Wei-Jie Zhang","doi":"10.1080/10826068.2025.2512946","DOIUrl":"https://doi.org/10.1080/10826068.2025.2512946","url":null,"abstract":"<p><p>Olive pomace, a by-product of olive oil production, remains underexplored despite its potential environmental and economic benefits. This study sequentially extracted three polysaccharides (OERC, OERH, and OERA) from olive pomace using water at room temperature, hot water (80 °C), and 2% Na<sub>2</sub>CO<sub>3</sub> solution (60 °C). Characterization through UV-Vis spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, thermogravimetric analysis, high-performance liquid chromatography, and Congo red experiment revealed that these polysaccharides are heteropolysaccharides primarily composed of glucose, mannose, rhamnose, and galactose, each with distinct molar ratios. The polysaccharides exhibited significant antioxidant activity by scavenging hydroxyl, DPPH, and superoxide radicals, with OERC showing the highest potency. They also repaired H<sub>2</sub>O<sub>2</sub>-induced oxidative damage in HepG2 cells, with OERC at 400 μg/mL and OERH/OERA at 200 μg/mL displaying optimal efficacy. Additionally, these polysaccharides effectively inhibited α-glucosidase activity, enhanced glucose consumption, and increased glycogen content in insulin-resistant models, thereby exerting hypoglycemic effects. Furthermore, they promoted proliferation, phagocytosis, and nitric oxide release in RAW264.7 macrophages, demonstrating immunomodulatory effects. These results indicate that sequential extraction under varying conditions is an effective method for preparing polysaccharides. The three polysaccharides isolated in this study show great potential for development as functional products with antioxidant, hypoglycemic, and immunomodulatory applications.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-17"},"PeriodicalIF":2.0,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144226460","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}
Zainitkumar A Dhameliya, Vilas P Chaudhari, Vishwa K Desai, Debesh R Roy
{"title":"Green synthesis of ZnO-NiO nanocomposite using <i>citrus sinensis</i> peel extract and their significant antibacterial properties against series of pathogenic strains.","authors":"Zainitkumar A Dhameliya, Vilas P Chaudhari, Vishwa K Desai, Debesh R Roy","doi":"10.1080/10826068.2025.2509894","DOIUrl":"https://doi.org/10.1080/10826068.2025.2509894","url":null,"abstract":"<p><p>This paper discusses the green synthesis of a ZnO-NiO nanocomposite using an extract of <i>citrus sinensis</i> peel as a capping and reducing agent. The green synthesized nanocomposite was characterized by X-ray Diffraction (XRD), Energy Dispersive X-ray Spectroscopy (EDX), Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR), and Ultraviolet-Visible Spectroscopy (UV-Vis) techniques. XRD is explored for crystallinity, and the crystalline size is found to be 36 nm for the nanocomposite. EDX is utilized for purity, and FTIR is used to establish the composition of the nanocomposites. SEM revealed that ZnO-NiO nanocomposites have a spherical-like surface morphology. The bandgap of the green synthesized nanocomposite is found to be 3.08 eV from reflectance data. By calculating the Zone of inhibition (ZOI) using the agar well diffusion method, the antibacterial activity (ABA) was investigated against three Gram-positive strains and five Gram-negative bacterial strains. The ZOI values for different bacterial strains ranges from 3 ± 0.5 mm to 16 ± 0.5 mm for a 75 µL nanocomposite concentration. The findings in the present work demonstrate that with an increase of the concentration of ZnO-NiO nanocomposite, their antibacterial activity (ABA) also increases, suggesting its merit in showing broad applicability for various uses in medicine.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-10"},"PeriodicalIF":2.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144199899","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}
Shabarni Gaffar, Rachelia Salsabila Zulfa, Fadhil Adikoesoemo, Syifa Tazkia, Korry Novitriani, Dani Permana, Sri Agung Fitri Kusuma, Hesti Lina Wiraswati, Mia Tria Novianti, Safri Ishmayana, Muhammad Yusuf, Toto Subroto
{"title":"Response surface methodology for optimization of scFv-BAD anti-E2 CHIKV expression in <i>Escherichia coli</i> Origami B (DE3) for the detection of Chikungunya virus.","authors":"Shabarni Gaffar, Rachelia Salsabila Zulfa, Fadhil Adikoesoemo, Syifa Tazkia, Korry Novitriani, Dani Permana, Sri Agung Fitri Kusuma, Hesti Lina Wiraswati, Mia Tria Novianti, Safri Ishmayana, Muhammad Yusuf, Toto Subroto","doi":"10.1080/10826068.2025.2512944","DOIUrl":"https://doi.org/10.1080/10826068.2025.2512944","url":null,"abstract":"<p><p>Chikungunya virus (CHIKV) is a mosquito-borne pathogen responsible for recurring outbreaks, highlighting the need for rapid and accurate diagnostic tools. The E2 glycoprotein of CHIKV is a promising target for antibody-based detection. This study aims to determine the optimal expression conditions of recombinant single-chain variable fragment (scFv) fused with biotin acceptor domain (BAD) specific to the CHIKV E2 glycoprotein in <i>Escherichia coli</i> Origami B (DE3). The optimization was performed using the Box-Behnken design of response surface methodology (RSM), with the IPTG inducer concentration, induction time, and induction temperature as the independent variables. The optimal conditions were identified as 0.2 mM IPTG, 2 hours induction at 37 °C, resulting in a total protein concentration of 0.658 mg/mL. The soluble fraction of scFv-BAD was successfully purified using Ni<sup>2+</sup>-NTA affinity chromatography, with a purity of 91.11%. ELISA confirmed that the recombinant scFv-BAD was biotinylated and retained its ability to bind the CHIKV E2 antigen. The optimized scFv-BAD construct demonstrates potential for use in various immunoassay platforms, including rapid diagnostic tests for CHIKV detection.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-11"},"PeriodicalIF":2.0,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144182015","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}