Inactivation of Coronaviruses during Sample Preparation for Proteomics Experiments

Area of Science:

• Virology
• Proteomics
• Biochemistry

Background:

• Mass spectrometry-based proteomics is crucial for SARS-CoV-2 research and diagnostics.
• Safe inactivation of coronaviruses in lysis buffers is essential for biosafety but not well-established.
• Current safety protocols often include a heating step, which could be omitted if buffers ensure inactivation.

Purpose of the Study:

• To systematically evaluate the coronavirus inactivation capacity of five common proteomics lysis buffers.
• To determine if lysis buffers can safely inactivate coronaviruses at room temperature, potentially eliminating the need for a heating step.

Main Methods:

• Tested five lysis buffers: 4% SDS, 1% SDC, TFA, 6 M GdmCl, and 8 M urea.
• Used human coronavirus 229E (HCoV-229E) and HCoV-OC43 as surrogates for SARS-CoV-2.
• Performed lysis at room temperature and 95°C for 5 minutes, confirming inactivation via cell culture (cytopathic effect) and quantitative real-time PCR.

Main Results:

• At room temperature, SDS, SDC, and TFA effectively inactivated both HCoV-229E and HCoV-OC43.
• GdmCl and urea showed incomplete inactivation at room temperature.
• Heating to 95°C likely enhances inactivation for all tested buffers.

Conclusions:

• The choice of lysis buffer significantly impacts coronavirus inactivation and biosafety in proteomics workflows.
• SDS, SDC, and TFA offer safe inactivation at room temperature, potentially streamlining sample preparation for SARS-CoV-2 analysis.
• Careful consideration of lysis buffer composition is critical to ensure biosafety during coronavirus proteomics studies and diagnostics.