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Harnessing sulfur-binding domains to separate Sp and Rp isomers of phosphorothioate oligonucleotides.
Appl Microbiol Biotechnol
August 2024
State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, 200240, People's Republic of China.
Chemical synthesis of phosphoromonothioate oligonucleotides (PS-ONs) is not stereo-specific and produces a mixture of Rp and Sp diastereomers, whose disparate reactivity can complicate applications. Although the current methods to separate these diastereomers which rely on chromatography are constantly improving, many Rp and Sp diastereomers are still co-eluted. Here, based on sulfur-binding domains that specifically recognize phosphorothioated DNA and RNA in Rp configuration, we developed a universal separation system for phosphorothioate oligonucleotide isomers using immobilized SBD (SPOIS).
View Article and Find Full Text PDFLatest Developments in Direct and Non-Direct LC-MS Methods Based on Liquid Electron Ionization (LEI).
Crit Rev Anal Chem
July 2024
Department of Pure and Applied Sciences, University of Urbino Carlo Bo, Urbino, Italy.
Mass spectrometry (MS) enables precise identification and quantification of molecules, particularly when combined with chromatography. The advent of atmospheric pressure ionization (API) techniques allowed the efficient coupling of liquid chromatography with MS (LC-MS), extending analyses to nonvolatile and thermolabile compounds. API techniques present limitations such as low informative capacity and reproducibility of mass spectra, increasing instrument complexity and costs.
View Article and Find Full Text PDFA Fast and Sensitive One-Tube SARS-CoV-2 Detection Platform Based on RTX-PCR and Argonaute.
Biosensors (Basel)
May 2024
State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei University, Wuhan 430062, China.
Since SARS-CoV-2 is a highly transmissible virus, alternative reliable, fast, and cost-effective methods are still needed to prevent virus spread that can be applied in the laboratory and for point-of-care testing. Reverse transcription real-time fluorescence quantitative PCR (RT-qPCR) is currently the gold criteria for detecting RNA viruses, which requires reverse transcriptase to reverse transcribe viral RNA into cDNA, and fluorescence quantitative PCR detection was subsequently performed. The frequently used reverse transcriptase is thermolabile; the detection process is composed of two steps: the reverse transcription reaction at a relatively low temperature, and the qPCR performed at a relatively high temperature, moreover, the RNA to be detected needs to pretreated if they had advanced structure.
View Article and Find Full Text PDFEcotoxicological assessments of atmospheric biomonitors exposed to urban pollution in a Brazilian metropolis.
Ecotoxicol Environ Saf
June 2024
Chemistry Department, Pontifical Catholic University of Rio de Janeiro, Marquês de São Vicente, 225, Gávea, Rio de Janeiro, Brazil. Electronic address:
Subcellular metal distribution assessments are the most adequate biomonitoring approach to evaluate metal toxicity, instead of total metal assessments This study aimed to assess subcellular metal distributions and associations to the main metal exposure biomarker, metallothionein (MT), in two bromeliad species (Tillandsia usneoides and Tillandsia stricta) exposed established in industrial, urban, and port areas in the metropolitan region of Rio de Janeiro, southeastern Brazil, through an active biomonitoring approach conducted one year. Metals and metalloids in three subcellular fractions (insoluble, thermolabile and thermostable) obtained from the MT purification process were determined by inductively coupled plasma mass spectrometry (ICP-MS). Lower MT concentrations were observed both during the dry sampling periods, associated to the crassulacean acid metabolism (CAM) and during the COVID-19 pandemic, due to reduced urban mobility, decreasing pollutant emissions.
View Article and Find Full Text PDFMicrowave-Dependent Thermo-Release Approach for Oligonucleotides 5'-Phosphorylation.
Org Lett
February 2024
Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznań, Poland.
A method for phosphorylating oligonucleotides using a thermosensitive "trigger" is hereby presented. The recovery of the phosphate specifically takes place under neutral conditions when subjected to an elevated temperature. Two identical thermolabile protecting groups are differentially removed with the initial release occurring swiftly and the second at a more gradual pace.
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