Development of a High-Throughput Mass Spectrometry-Based SARS-CoV-2 Immunoassay.

Area of Science:

• Biomedical Diagnostics
• Analytical Chemistry
• Mass Spectrometry

Background:

• The COVID-19 pandemic highlighted the need for advanced diagnostic methods that are rapid, reliable, and high-throughput.
• Current diagnostic approaches face limitations in patient-friendliness, demand capacity, and result consistency.

Purpose of the Study:

• To develop a novel diagnostic strategy for on-demand detection of infectious agents.
• To address the limitations of existing diagnostic technologies for widespread and reliable testing.

Main Methods:

• A two-point separation strategy employing laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS).
• Utilization of a stable, laser-cleavable ionic probe as a mass reporter conjugated to antibodies.
• Quantitative analysis using a laser-cleavable internal standard for accurate measurements.

Main Results:

• Demonstrated ultrasensitive detection (amol level) of the SARS-CoV-2 Spike S1 subunit antibody without amplification.
• Achieved high-throughput, interruptible, storable, and restorable on-demand detection capabilities.
• Successfully identified positive and negative samples containing the spike protein.

Conclusions:

• The proposed LDI-TOF MS strategy with a cleavable ionic probe offers a sensitive and versatile platform for rapid diagnostics.
• This approach is applicable for detecting various disease antigens beyond SARS-CoV-2.
• The method enhances diagnostic capabilities for infectious diseases, meeting critical public health needs.