Engineered Receptor Capture Combined with Mass Spectrometry Enables High-Throughput Detection and Quantitation of SARS-CoV-2 Spike Protein

JACS Au, Journal Year: 2025, Volume and Issue: 5(2), P. 747 - 755

Published: Feb. 4, 2025

Mass spectrometry (MS) is a potentially powerful approach for the diagnostic detection of SARS-CoV-2 and other viruses. However, MS compromised when viral antigens are present at low concentrations, especially in complex biological media. We hypothesized that receptors could be used target capture to enable by under such conditions. This was tested using extracellular domain receptor ACE2. To maximize recovery protein, directed protein evolution first increase affinity ACE2 spike protein. generated an evolved with increased binding receptor-binding (RBD). as affinity-enhanced forms ACE2, sensitive mutations variant RBDs. As alternative strategy capture, native engineered addition oligomerization scaffold create pentameric bound extremely tightly RBD, apparent several thousand-fold over monomeric RBD retention more than 8 h. Immobilization multimeric enabled quantitative enrichment from saliva sensitivity MS. These data show combined can effective, rapid method quantitation A similar attachment proteins viruses or any which there suitable receptors.

Language: Английский

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Mass spectrometry-based ligand binding assays in biomedical research

Expert Review of Proteomics, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 18, 2025

Ligand binding assays combining immunoaffinity enrichment steps with mass spectrometry (MS) readout have gained attention as a highly specific and sensitive tool for protein quantification. These techniques typically combine enzymatic fragmentation of the sample or enriched capture on peptide-level Antibodies ensure target recognition, while MS offers quantitative accuracy isotopically labeled internal standards. This dual approach supports broad dynamic range, enabling measurements from picomolar to nanomolar levels. methods diverse applications, quantifying signaling proteins in basic research biomarker monitoring clinical trials analyzing pharmacokinetics therapeutic proteins. review delves into workflows immunoaffinity-MS, shedding light innovative strategies employed, their practical efficacy, inherent limitations realm Immunoaffinity-MS has transformed analysis, but widespread adoption is hindered by complex workflows, high instrument costs, limited molecule availability. Efforts enhance automation, standardize advance technological innovation aim overcome these barriers. Improvements spectrometer sensitivity, advances recombinant technologies, support public initiatives are poised further improve reliability accessibility this method.

Language: Английский

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SARS-CoV-2 is here to stay: do not lower our guard

Clinical Chemistry and Laboratory Medicine (CCLM), Journal Year: 2024, Volume and Issue: 62(6), P. 1017 - 1018

Published: March 26, 2024

Language: Английский

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Immunocapture mass spectrometry: macroscopic history, recent trends and future prospects

Bioanalysis, Journal Year: 2024, Volume and Issue: unknown, P. 1 - 10

Published: Oct. 17, 2024

Literature on immunological pretreatment and mass spectrometry was searched in the Web of Science database. The collected data were utilized for bibliometric analysis literature review to identify trends this field. Immunological initially began chemical toxicology using gas chromatography research interests later shifted biologics pharmacokinetics, biomarkers, anti-drug antibodies diagnosis. key advantage ability distinguish molecular subtypes. Future are expected focus a broader range analyte types (such as intact protein spectrometry) methodological improvements, including better digestion microfluidics, aiming faster, less labor-intensive more reliable through multiplexing automation.

Language: Английский

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