ACS Bio & Med Chem Au最新文献_第2页

Innovative Dual Combination Cospray-Dried Rock Inhibitor/l-Carnitine Inhalable Dry Powder Aerosols

IF 3.8

ACS Bio & Med Chem Au Pub Date : 2024-10-28 DOI: 10.1021/acsbiomedchemau.4c0006310.1021/acsbiomedchemau.4c00063

Maria F. Acosta, David Encinas-Basurto, Michael D. Abrahamson, Basanth Babu Eedara, Don Hayes Jr., Jeffrey R. Fineman, Stephen M. Black and Heidi M. Mansour*,

{"title":"Innovative Dual Combination Cospray-Dried Rock Inhibitor/l-Carnitine Inhalable Dry Powder Aerosols","authors":"Maria F. Acosta, David Encinas-Basurto, Michael D. Abrahamson, Basanth Babu Eedara, Don Hayes Jr., Jeffrey R. Fineman, Stephen M. Black and Heidi M. Mansour*, ","doi":"10.1021/acsbiomedchemau.4c0006310.1021/acsbiomedchemau.4c00063","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00063https://doi.org/10.1021/acsbiomedchemau.4c00063","url":null,"abstract":"This study introduces novel cospray-dried (Co-SD) formulations of simvastatin, a Nrf2 activator ROCK inhibitor, with l-carnitine as molecular mixtures in various molar ratios for targeted pulmonary inhalation aerosol delivery in pulmonary hypertension, optimized for excipient-free dry powder inhalers (DPIs). The two components were spray-dried at various molar ratios by using different starting feed solution concentrations and process parameters. In addition to comprehensive physicochemical characterization, in vitro aerosol dispersion performance as DPIs using two FDA-approved DPI devices with different shear stress properties, in vitro viability as a function of dose on 2D human pulmonary cellular monolayers and on 3D small airway epithelia human primary cultures at the air–liquid interface (ALI), and in vitro transepithelial electrical resistance (TEER) at the ALI were conducted. Solid-state physicochemical characterization confirmed homogeneous molecular mixtures and the crystalline nature of the Co-SD formulations. In vitro aerosolization dispersion performance demonstrated that all Co-SD dual combination molecular mixtures aerosolized successfully with both human FDA-approved DPI devices, had ∼100% emitted dose, and good fine particle fraction values. The in vitro viability and TEER assays demonstrated that all formulations were safe to the human pulmonary cell as 2D and 3D cultures as a function of dose.","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"300–318 300–318"},"PeriodicalIF":3.8,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00063","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142844190","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}

引用次数: 0

Innovative Dual Combination Cospray-Dried Rock Inhibitor/l-Carnitine Inhalable Dry Powder Aerosols.

IF 3.8

ACS Bio & Med Chem Au Pub Date : 2024-10-28 eCollection Date: 2024-12-18 DOI: 10.1021/acsbiomedchemau.4c00063

Maria F Acosta, David Encinas-Basurto, Michael D Abrahamson, Basanth Babu Eedara, Don Hayes, Jeffrey R Fineman, Stephen M Black, Heidi M Mansour

{"title":"Innovative Dual Combination Cospray-Dried Rock Inhibitor/l-Carnitine Inhalable Dry Powder Aerosols.","authors":"Maria F Acosta, David Encinas-Basurto, Michael D Abrahamson, Basanth Babu Eedara, Don Hayes, Jeffrey R Fineman, Stephen M Black, Heidi M Mansour","doi":"10.1021/acsbiomedchemau.4c00063","DOIUrl":"10.1021/acsbiomedchemau.4c00063","url":null,"abstract":"This study introduces novel cospray-dried (Co-SD) formulations of simvastatin, a Nrf2 activator ROCK inhibitor, with l-carnitine as molecular mixtures in various molar ratios for targeted pulmonary inhalation aerosol delivery in pulmonary hypertension, optimized for excipient-free dry powder inhalers (DPIs). The two components were spray-dried at various molar ratios by using different starting feed solution concentrations and process parameters. In addition to comprehensive physicochemical characterization, in vitro aerosol dispersion performance as DPIs using two FDA-approved DPI devices with different shear stress properties, in vitro viability as a function of dose on 2D human pulmonary cellular monolayers and on 3D small airway epithelia human primary cultures at the air-liquid interface (ALI), and in vitro transepithelial electrical resistance (TEER) at the ALI were conducted. Solid-state physicochemical characterization confirmed homogeneous molecular mixtures and the crystalline nature of the Co-SD formulations. In vitro aerosolization dispersion performance demonstrated that all Co-SD dual combination molecular mixtures aerosolized successfully with both human FDA-approved DPI devices, had ∼100% emitted dose, and good fine particle fraction values. The in vitro viability and TEER assays demonstrated that all formulations were safe to the human pulmonary cell as 2D and 3D cultures as a function of dose.","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"300-318"},"PeriodicalIF":3.8,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659894/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142877683","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}

引用次数: 0

Structural Evidence for DUF512 as a Radical S-Adenosylmethionine Cobalamin-Binding Domain

IF 3.8

ACS Bio & Med Chem Au Pub Date : 2024-10-23 DOI: 10.1021/acsbiomedchemau.4c0006710.1021/acsbiomedchemau.4c00067

Bo Wang, Amy E. Solinski, Matthew I. Radle, Olivia M. Peduzzi, Hayley L. Knox, Jiayuan Cui, Ravi K. Maurya, Neela H. Yennawar and Squire J. Booker*,

{"title":"Structural Evidence for DUF512 as a Radical S-Adenosylmethionine Cobalamin-Binding Domain","authors":"Bo Wang, Amy E. Solinski, Matthew I. Radle, Olivia M. Peduzzi, Hayley L. Knox, Jiayuan Cui, Ravi K. Maurya, Neela H. Yennawar and Squire J. Booker*, ","doi":"10.1021/acsbiomedchemau.4c0006710.1021/acsbiomedchemau.4c00067","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00067https://doi.org/10.1021/acsbiomedchemau.4c00067","url":null,"abstract":"Cobalamin (Cbl)-dependent radical S-adenosylmethionine (SAM) enzymes constitute a large subclass of radical SAM (RS) enzymes that use Cbl to catalyze various types of reactions, the most common of which are methylations. Most Cbl-dependent RS enzymes contain an N-terminal Rossmann fold that aids Cbl binding. Recently, it has been demonstrated that the methanogenesis marker protein 10 (Mmp10) requires Cbl to methylate an arginine residue in the α-subunit of methyl coenzyme M reductase. However, Mmp10 contains a Cbl-binding domain in the C-terminal region of its primary structure that does not share significant sequence similarity with canonical RS Cbl-binding domains. Bioinformatic analysis of Mmp10 identified DUF512 (Domain of Unknown Function 512) as a potential Cbl-binding domain in RS enzymes. In this paper, four randomly selected DUF512-containing proteins from various organisms were overexpressed, purified, and shown to bind Cbl. X-ray crystal structures of DUF512-containing proteins from Clostridium sporogenes and Pyrococcus furiosus were determined, confirming their C-terminal Cbl-binding domains. The structure of the DUF512-containing protein from C. sporogenes is the first of an RS enzyme containing a PDZ domain. Its RS domain has an unprecedented β3α4 core, whereas most RS enzymes adopt a (βα)6 core. The DUF512-containing protein from P. furiosus has no PDZ domain, but its RS domain also has an uncommon (βα)5 core.","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"319–330 319–330"},"PeriodicalIF":3.8,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00067","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142844037","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}

引用次数: 0

Structural Evidence for DUF512 as a Radical S-Adenosylmethionine Cobalamin-Binding Domain.

IF 3.8

ACS Bio & Med Chem Au Pub Date : 2024-10-23 eCollection Date: 2024-12-18 DOI: 10.1021/acsbiomedchemau.4c00067

Bo Wang, Amy E Solinski, Matthew I Radle, Olivia M Peduzzi, Hayley L Knox, Jiayuan Cui, Ravi K Maurya, Neela H Yennawar, Squire J Booker

{"title":"Structural Evidence for DUF512 as a Radical S-Adenosylmethionine Cobalamin-Binding Domain.","authors":"Bo Wang, Amy E Solinski, Matthew I Radle, Olivia M Peduzzi, Hayley L Knox, Jiayuan Cui, Ravi K Maurya, Neela H Yennawar, Squire J Booker","doi":"10.1021/acsbiomedchemau.4c00067","DOIUrl":"10.1021/acsbiomedchemau.4c00067","url":null,"abstract":"Cobalamin (Cbl)-dependent radical S-adenosylmethionine (SAM) enzymes constitute a large subclass of radical SAM (RS) enzymes that use Cbl to catalyze various types of reactions, the most common of which are methylations. Most Cbl-dependent RS enzymes contain an N-terminal Rossmann fold that aids Cbl binding. Recently, it has been demonstrated that the methanogenesis marker protein 10 (Mmp10) requires Cbl to methylate an arginine residue in the α-subunit of methyl coenzyme M reductase. However, Mmp10 contains a Cbl-binding domain in the C-terminal region of its primary structure that does not share significant sequence similarity with canonical RS Cbl-binding domains. Bioinformatic analysis of Mmp10 identified DUF512 (Domain of Unknown Function 512) as a potential Cbl-binding domain in RS enzymes. In this paper, four randomly selected DUF512-containing proteins from various organisms were overexpressed, purified, and shown to bind Cbl. X-ray crystal structures of DUF512-containing proteins from Clostridium sporogenes and Pyrococcus furiosus were determined, confirming their C-terminal Cbl-binding domains. The structure of the DUF512-containing protein from C. sporogenes is the first of an RS enzyme containing a PDZ domain. Its RS domain has an unprecedented β3α4 core, whereas most RS enzymes adopt a (βα)6 core. The DUF512-containing protein from P. furiosus has no PDZ domain, but its RS domain also has an uncommon (βα)5 core.","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"319-330"},"PeriodicalIF":3.8,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659888/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142877684","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}

引用次数: 0

An Investigation of Nirmatrelvir (Paxlovid) Resistance in SARS-CoV-2 Mpro

IF 3.8

ACS Bio & Med Chem Au Pub Date : 2024-10-08 DOI: 10.1021/acsbiomedchemau.4c0004510.1021/acsbiomedchemau.4c00045

Rasha M. Yaghi, Dennis C. Wylie, Collin L. Andrews, Olivia H. Dickert, Anjana Ram and Brent L. Iverson*,

{"title":"An Investigation of Nirmatrelvir (Paxlovid) Resistance in SARS-CoV-2 Mpro","authors":"Rasha M. Yaghi, Dennis C. Wylie, Collin L. Andrews, Olivia H. Dickert, Anjana Ram and Brent L. Iverson*, ","doi":"10.1021/acsbiomedchemau.4c0004510.1021/acsbiomedchemau.4c00045","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00045https://doi.org/10.1021/acsbiomedchemau.4c00045","url":null,"abstract":"The high throughput YESS 2.0 platform was used to screen a large library of SARS-CoV-2 Mpro variants in the presence of nirmatrelvir. Of the 100 individual most prevalent mutations identified in the screen and reported here, the most common were E166V, L27V, N142S, A173V, and Y154N, along with their various combinations. In vitro analysis revealed that resistance to nirmatrelvir for these individual mutations, as well as all of the combinations we analyzed, was accompanied by decreased catalytic activity with the native substrate. Importantly, the mutations we identified have not appeared as significantly enriched in SARS-CoV-2 Mpro sequences isolated from COVID-19 patients following the introduction of nirmatrelvir. We also analyzed three of the most common SARS-CoV-2 Mpro mutations that have been seen in patients recently, and only a measured increase in nirmatrelvir resistance was seen when the more recently appearing A285V is added to both P132H and K90R. Taken together, our results predict that resistance to nirmatrelvir will be slower to develop than expected based on experience with other viral protease inhibitors, perhaps due in part to the close structural correspondence between nirmatrelvir and SARS-CoV-2 Mpro’s preferred substrates.","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"280–290 280–290"},"PeriodicalIF":3.8,"publicationDate":"2024-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00045","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142844146","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}

引用次数: 0

An Investigation of Nirmatrelvir (Paxlovid) Resistance in SARS-CoV-2 M^pro.

IF 3.8

ACS Bio & Med Chem Au Pub Date : 2024-10-08 eCollection Date: 2024-12-18 DOI: 10.1021/acsbiomedchemau.4c00045

Rasha M Yaghi, Dennis C Wylie, Collin L Andrews, Olivia H Dickert, Anjana Ram, Brent L Iverson

{"title":"An Investigation of Nirmatrelvir (Paxlovid) Resistance in SARS-CoV-2 Mpro.","authors":"Rasha M Yaghi, Dennis C Wylie, Collin L Andrews, Olivia H Dickert, Anjana Ram, Brent L Iverson","doi":"10.1021/acsbiomedchemau.4c00045","DOIUrl":"10.1021/acsbiomedchemau.4c00045","url":null,"abstract":"The high throughput YESS 2.0 platform was used to screen a large library of SARS-CoV-2 Mpro variants in the presence of nirmatrelvir. Of the 100 individual most prevalent mutations identified in the screen and reported here, the most common were E166V, L27V, N142S, A173V, and Y154N, along with their various combinations. In vitro analysis revealed that resistance to nirmatrelvir for these individual mutations, as well as all of the combinations we analyzed, was accompanied by decreased catalytic activity with the native substrate. Importantly, the mutations we identified have not appeared as significantly enriched in SARS-CoV-2 Mpro sequences isolated from COVID-19 patients following the introduction of nirmatrelvir. We also analyzed three of the most common SARS-CoV-2 Mpro mutations that have been seen in patients recently, and only a measured increase in nirmatrelvir resistance was seen when the more recently appearing A285V is added to both P132H and K90R. Taken together, our results predict that resistance to nirmatrelvir will be slower to develop than expected based on experience with other viral protease inhibitors, perhaps due in part to the close structural correspondence between nirmatrelvir and SARS-CoV-2 Mpro's preferred substrates.","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"280-290"},"PeriodicalIF":3.8,"publicationDate":"2024-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659887/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142878252","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}

引用次数: 0

New Catalytic Residues and Catalytic Mechanism of the RNase T1 Family RNase T1 家族的新催化残基和催化机理

ACS Bio & Med Chem Au Pub Date : 2024-09-20 DOI: 10.1021/acsbiomedchemau.4c00046

Katsuki Takebe, Mamoru Suzuki, Yumiko Hara, Takuya Katsutani, Naomi Motoyoshi, Tadashi Itagaki, Shuhei Miyakawa, Kuniaki Okamoto, Kaori Fukuzawa, Hiroko Kobayashi

引用次数: 0

Design, Synthesis, and Biological Evaluation of Darunavir Analogs as HIV-1 Protease Inhibitors 作为 HIV-1 蛋白酶抑制剂的 Darunavir 类似物的设计、合成和生物学评价

ACS Bio & Med Chem Au Pub Date : 2024-09-19 DOI: 10.1021/acsbiomedchemau.4c00040

Muhammad Asad Ur Rehman, Hathaichanok Chuntakaruk, Soraat Amphan, Aphinya Suroengrit, Kowit Hengphasatporn, Yasuteru Shigeta, Thanyada Rungrotmongkol, Kuakarun Krusong, Siwaporn Boonyasuppayakorn, Chanat Aonbangkhen, Tanatorn Khotavivattana

{"title":"Design, Synthesis, and Biological Evaluation of Darunavir Analogs as HIV-1 Protease Inhibitors","authors":"Muhammad Asad Ur Rehman, Hathaichanok Chuntakaruk, Soraat Amphan, Aphinya Suroengrit, Kowit Hengphasatporn, Yasuteru Shigeta, Thanyada Rungrotmongkol, Kuakarun Krusong, Siwaporn Boonyasuppayakorn, Chanat Aonbangkhen, Tanatorn Khotavivattana","doi":"10.1021/acsbiomedchemau.4c00040","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00040","url":null,"abstract":"Darunavir, a frontline treatment for HIV infection, faces limitations due to emerging multidrug resistant (MDR) HIV strains, necessitating the development of analogs with improved activity. In this study, a combinatorial in silico approach was used to initially design a series of HIV-1 PI analogs with modifications at key sites, P1′ and P2′, to enhance interactions with HIV-1 PR. Fifteen analogs with promising binding scores were selected for synthesis and evaluated for the HIV-1 PR inhibition activity. The variation of P2′ substitution was found to be effective, as seen in 5aa (1.54 nM), 5ad (0.71 nM), 5ac (0.31 nM), 5ae (0.28 nM), and 5af (1.12 nM), featuring halogen, aliphatic, and alkoxy functionalities on the phenyl sulfoxide motif exhibited superior inhibition against HIV-1 PR compared to DRV, with minimal cytotoxicity observed in Vero and 293T cell lines. Moreover, computational studies demonstrated the potential of selected analogs to inhibit various HIV-1 PR mutations, including I54M and I84V. Further structural dynamics and energetic analyses confirmed the stability and binding affinity of promising analogs, particularly 5ae, which showed strong interactions with key residues in HIV-1 PR. Overall, this study underscores the importance of flexible moieties and interaction enhancement at the S2′ subsite of HIV-1 PR in developing effective DRV analogs to combat HIV and other global health issues.","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"22 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142257770","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}

引用次数: 0

Directed Evolution of Candidatus Methanomethylophilus alvus Pyrrolysyl-tRNA Synthetase for the Genetic Incorporation of Two Different Noncanonical Amino Acids in One Protein 嗜甲烷甲酵母菌（Candidatus Methanomethylophilus alvus）吡咯氨酰-tRNA 合成酶的定向进化，在一个蛋白质中遗传性地加入两种不同的非顺式氨基酸

ACS Bio & Med Chem Au Pub Date : 2024-08-22 DOI: 10.1021/acsbiomedchemau.4c00028

Chia-Chuan D. Cho, Waye Michelle Leeuwon, Wenshe Ray Liu

{"title":"Directed Evolution of Candidatus Methanomethylophilus alvus Pyrrolysyl-tRNA Synthetase for the Genetic Incorporation of Two Different Noncanonical Amino Acids in One Protein","authors":"Chia-Chuan D. Cho, Waye Michelle Leeuwon, Wenshe Ray Liu","doi":"10.1021/acsbiomedchemau.4c00028","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00028","url":null,"abstract":"The genetic code expansion technique is a powerful chemical biology tool to install noncanonical amino acids (ncAAs) in proteins. As a key enzyme for this technique, pyrrolysyl-tRNA synthetase (PylRS), coupled with its cognate amber suppressor tRNAPyl, has been engineered for the genetic incorporation of more than 200 ncAAs. Using PylRS clones from different archaeal origins, two ncAAs have also been genetically encoded in one protein. In this work, we show that the C41AU mutant of tRNAPyl from Candidatus Methanomethylophilus alvus (CmatRNAPyl) is catalytically inert toward PylRS from Methanosarcina mazei (MmPylRS) but has weak activity toward PylRS from Ca. M. alvus (CmaPylRS). To improve the catalytic efficiency of CmaPylRS toward CmatRNAPyl-C41AU, we conducted a directed evolution of CMaPylRS by randomizing its coding sequence, followed by the screening of active mutant clones. After three rounds of randomization and screening, we identified 4 mutations, Y16F/N57D/E161G/N182I, that improve the catalytic efficiency of CMaPylRS toward CMatRNAPyl-C41AU. This new clone, named R3–14, coupling with CmatRNAPyl-C41AU to recognize an amber codon, has been successfully used together with an evolved MmPylRS clone, coupling with a mutant M. mazei tRNAPyl to recognize an ochre codon, to genetically incorporate two different ncAAs, Nε-(t-butoxycarbonyl)-lysine and Nε-acetyl-lysine, into one model protein.","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"2674 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142204333","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}

引用次数: 0

Development of a Polymersome Blood Ammonia Assay Coupled with a Portable Near-Infrared Fluorometer 开发与便携式近红外荧光测定仪相结合的聚合体血氨测定法

ACS Bio & Med Chem Au Pub Date : 2024-07-30 DOI: 10.1021/acsbiomedchemau.4c00013

Marie-Lynn Al-Hawat, Justine Caron, Sarah Djebbar, Simon Matoori

{"title":"Development of a Polymersome Blood Ammonia Assay Coupled with a Portable Near-Infrared Fluorometer","authors":"Marie-Lynn Al-Hawat, Justine Caron, Sarah Djebbar, Simon Matoori","doi":"10.1021/acsbiomedchemau.4c00013","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00013","url":null,"abstract":"Ammonia is a key biomarker in inborn and acquired liver disease. As clinical point-of-care blood ammonia assays are lacking, we developed a polymersome formulation for point-of-care blood ammonia sensing combined with a portable fluorometer. A pH-sensitive near-infrared (NIR) fluorescent dye was identified, which showed a strong fluorescence increase at acidic pH values. Building on reports on ammonia-selective PS-b-PEG polymersomes, these polymersomes were loaded with the NIR dye. These NIR fluorescent polymersomes sensed ammonia in a clinically relevant range in ammonia-spiked fresh whole blood with high linearity (R2 = 0.9948) after 5 min using a conventional tabletop plate reader. Subsequently, the assay was tested with a portable fluorometer. An ammonia-dependent fluorescence increase was detected in ammonia-spiked fresh mouse blood after 5 min using the portable fluorometer. The NIR dye-loaded PS-b-PEG polymersomes rapidly sensed ammonia with high linearity in whole blood. This assay was successfully combined with a portable fluorometer and only required 3 μL of blood. These findings motivate a further development and clinical translation of this point-of-care blood ammonia assay.","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"16 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141870566","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}

引用次数: 0