Rationale: The Q Orbitrap permits multiplexed targeted selected ion monitoring (SIM) or tandem mass spectrometry (MS/MS) scans. Such scans provide a significantly higher sensitivity than conventional full scan acquisition. However, due to the multiplexing, a monitored product ion extracted from a MS/MS scan can no longer be linked to the precursor ion from which it was derived. Furthermore, due to the automatic gain control, quantification based on targeted product ions acquired in the multiplexed MS/MS mode can become questionable.
Methods: The application programming interface (API) of the Q Exactive has been used to program a multiplexed targeted MS/MS mode that permits the establishment of a link (based on digital coding) between the product ion and the responsible precursor ion. Furthermore, switching off the automatic gain control feature and the definition of appropriate C-trap settings were tested to improve the quantification performance.
Results: The use of dedicated decoding scans permitted the clear assignment of all monitored product ions to the responsible precursor ion. Furthermore, the customized multiplexed targeted scan used for quantification showed good sensitivity and linearity for a maximum number of eight co-eluting analytes.
Conclusions: Multiplexed targeted MS/MS scans can be optimized to provide better selectivity (correct linking of an observed product ion to the responsible precursor ion) as well as improved quantitative performance (enforcement of an identical ion injection time for all targeted precursor ions). These two improvements are relevant for quantitative residue analysis.
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http://dx.doi.org/10.1002/rcm.7532 | DOI Listing |
Tertiary lymphoid structures (TLS) are organized immune cell aggregates that arise in chronic inflammatory conditions. In cancer, TLS are associated with better prognosis and enhanced response to immunotherapy, making these structures attractive therapeutic targets. However, the mechanisms regulating TLS formation and maintenance in cancer are incompletely understood.
View Article and Find Full Text PDFClin Genet
January 2025
Prenatal Diagnosis and Fetal Medicine Department, Human Genetics and Genome Research Institute, National Research Centre (NRC), Cairo, Egypt.
SUMOylation involves covalent attachment of small ubiquitin-like modifier (SUMO) proteins to specific lysine residues on target proteins and regulates various aspects of their function. Sentrin-specific proteases (SENPs) are key players in both the conjugation reaction of SUMO proteins to their targets and the subsequent deconjugation of SUMO-conjugated substrates. Here, we provide the first comprehensive prenatal description of a lethal syndrome linked to a novel homozygous stop-gain variant in SENP7 c.
View Article and Find Full Text PDFBMC Ophthalmol
January 2025
Department of Tuberculosis, New District Branch of Northern Jiangsu People's Hospital of Jiangsu Province, Yangzhou, 225001, Jiangsu Province, China.
Background: This study aims to detect Mycobacterium tuberculosis complex (MTBC) DNA in intraocular fluid from clinically suspected tuberculous uveitis patients using multiplex polymerase chain reaction (PCR) and investigate the diagnostic utility of multiplex PCR for tuberculous uveitis.
Methods: Primers targeting three specific genes (MPB64, CYP141, and IS6110) within the MTBC genome were designed. Multiplex PCR was conducted using DNA from the H37Rv strain as well as DNA extracted from fluids of confirmed tuberculosis patients to assess primer specificity and method feasibility.
Mol Syst Biol
January 2025
Department of Pathology and Cell Biology, Columbia University Vagelos College of Physicians and Surgeons, New York, NY, 10032, USA.
With current treatments addressing only a fraction of pathogens and new viral threats constantly evolving, there is a critical need to expand our existing therapeutic arsenal. To speed the rate of discovery and better prepare against future threats, we establish a high-throughput platform capable of screening compounds against 40 diverse viral proteases simultaneously. This multiplex approach is enabled by using cellular biosensors of viral protease activity combined with DNA-barcoding technology, as well as several design innovations that increase assay sensitivity and correct for plate-to-plate variation.
View Article and Find Full Text PDFACS Infect Dis
January 2025
Department of Microbiology, Genetics, and Immunology, Michigan State University, East Lansing, Michigan 48824, United States.
Group B (GBS) is a major cause of fetal and neonatal mortality worldwide. Many of the adverse effects of invasive GBS are associated with inflammation; therefore, understanding bacterial factors that promote inflammation is of critical importance. Membrane vesicles (MVs), which are produced by many bacteria, may modulate host inflammatory responses.
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