{"title":"Gold Nanoparticle-Assisted CRISPR-Cas12a-Based Activity Assay for Highly Sensitive Detection of Trypsin.","authors":"Sathishkumar Munusamy, Haiyan Zheng, Jun Chen, Shuo Zhou, Juanhua Kong, Rana Jahani, Yuan Zhao, Xiyun Guan","doi":"10.1021/acsabm.5c00854","DOIUrl":"https://doi.org/10.1021/acsabm.5c00854","url":null,"abstract":"<p><p>Proteases play important roles in diverse physiological processes, and their malfunction has been implicated in various conditions and diseases. Therefore, development of sensitive methods for protease detection in clinical samples is highly desired for disease diagnosis. Herein, we report an ultrasensitive and selective CRISPR-Cas12a based fluorescent assay for trypsin activity measurement. By taking advantage of the signal amplification brought by a unique magnetic bead-gold nanoparticle assembly, which carries the peptide substrate and tens to hundreds of DNA molecules per peptide molecule, trypsin can be detected with a limit of detection reaching as low as 0.13 ng/mL. Furthermore, the sensor selectivity study was performed by examining several biomolecules commonly present in biological samples, including bovine serum albumin (BSA), human serum albumin (HSA), DNase, RNase, chymotrypsin, elastase, and thrombin. Moreover, trypsin inhibition and serum sample analysis were successfully carried out. Given the ultrahigh sensitivity, the CRISPR-based trypsin activity assay developed in this work can be used as a generic platform for developing sensors for other proteases, offering the potential as a noninvasive/minimally invasive tool for clinical diagnosis.</p>","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":" ","pages":""},"PeriodicalIF":4.6,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144309235","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"","authors":"Archana, Anil Kumar and Pratima R Solanki*, ","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":"8 6","pages":"XXX-XXX XXX-XXX"},"PeriodicalIF":4.6,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsabm.5c00329","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144357234","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Eskil André Karlsen, Mattias Berglin, Adam Hansson, Anders Oskar Lundgren and John S. M. Svendsen*,
{"title":"","authors":"Eskil André Karlsen, Mattias Berglin, Adam Hansson, Anders Oskar Lundgren and John S. M. Svendsen*, ","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":"8 6","pages":"XXX-XXX XXX-XXX"},"PeriodicalIF":4.6,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsabm.5c00253","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144357241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kameel Zuniga*, Marc Thompson, Preeti J. Muire, Nicholas Clay, S. L. Rajasekhar Karna, Logan Leatherman, Rebecca Lopez, Tao You, Krystle Harm, Jerod Brammer, Joseph Wenke, Robert Christy and Christine Kowalczewski,
{"title":"","authors":"Kameel Zuniga*, Marc Thompson, Preeti J. Muire, Nicholas Clay, S. L. Rajasekhar Karna, Logan Leatherman, Rebecca Lopez, Tao You, Krystle Harm, Jerod Brammer, Joseph Wenke, Robert Christy and Christine Kowalczewski, ","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":"8 6","pages":"XXX-XXX XXX-XXX"},"PeriodicalIF":4.6,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsabm.5c00100","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144425882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Soumya Saroj, Akshaya Kana Veedu, Chandrasekhar Reddy U, Nalini Venkatesan, Abhishek K. Verma and Muraleedharan Kannoth Manheri*,
{"title":"","authors":"Soumya Saroj, Akshaya Kana Veedu, Chandrasekhar Reddy U, Nalini Venkatesan, Abhishek K. Verma and Muraleedharan Kannoth Manheri*, ","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":"8 6","pages":"XXX-XXX XXX-XXX"},"PeriodicalIF":4.6,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsabm.4c01932","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144425898","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Qiuling Deng, Ziyu Huang, Minjin Wu, Xiaoping Wei, Jianping Li
{"title":"Ultrasensitive Electrochemiluminescence Immunosensor of Procalcitonin Based on Synergistic Enhancement by Hf-MOF/g-C<sub>3</sub>N<sub>4</sub> via Integration of Dual Luminophores, a Coreactant Accelerator, and Energy Transfer.","authors":"Qiuling Deng, Ziyu Huang, Minjin Wu, Xiaoping Wei, Jianping Li","doi":"10.1021/acsabm.5c00315","DOIUrl":"10.1021/acsabm.5c00315","url":null,"abstract":"<p><p>A synergistic enhanced electrochemiluminescence (ECL) method was proposed. Synergistic enhancement was achieved by incorporating a coreactant accelerator through energy transfer between dual luminophores, g-C<sub>3</sub>N<sub>4</sub> and Hf-MOF. In addition, a sandwich-type ECL immunosensor was developed for the ultrasensitive detection of procalcitonin (PCT), in which the primary antibody was labeled with the g-C<sub>3</sub>N<sub>4</sub>@Au nanoparticles that immobilized on an electrode surface and the secondary antibody was labeled with Hf-MOF. Apart from a luminophore, Hf-MOF also acted as a coreactant accelerator, catalyzing the generation of sulfate radical anions (SO<sub>4</sub><sup>•-</sup>) by reactive oxygen and potassium persulfate. Furthermore, Hf-MOF acted as an acceptor of energy transfer of the luminophore intermediate g-C<sub>3</sub>N<sub>4</sub>, which greatly enhances the ECL signal. The ECL intensities of Hf-MOF/g-C<sub>3</sub>N<sub>4</sub>/S<sub>2</sub>O<sub>8</sub><sup>2-</sup> were 2.3 and 4.4 times higher than the ECL intensities of the Hf-MOF/S<sub>2</sub>O<sub>8</sub><sup>2-</sup> and g-C<sub>3</sub>N<sub>4</sub>/S<sub>2</sub>O<sub>8</sub><sup>2-</sup> systems, respectively. The response signal of the ECL immunosensor showed a good linear relationship with 10 fg/mL-100 ng/mL PCT, showing a detection limit of 3.3 fg/mL, which was much lower than of the reported method. The proposed method is feasible for the rapid identification and detection of systemic bacterial infections and for the prevention of abuse of antibiotics.</p>","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":" ","pages":"4937-4945"},"PeriodicalIF":4.6,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144100958","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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