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Related Concept Videos

Conjugated Proteins02:50

Conjugated Proteins

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Simple proteins and protein complexes contain only amino acids. In contrast, many other proteins, called conjugated proteins, covalently bond with non-protein moieties.
Nucleoproteins are protein complexes that contain nucleic acids, categorized as deoxyribonucleoproteins (DNPs) or ribonucleoproteins (RNPs) respectively. The nucleosome is a typical example of a DNP where nuclear DNA is associated with histone proteins. The major antigen for the Covid-19 virus SARS-CoV is an RNP that is critical...
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The function of proteins depends on their native three-dimensional structure, which is dictated by the amino acid sequence of the specific protein. Folding of the polypeptide chain takes place under specific conditions that energetically favor the folded conformation. In contrast, protein denaturation occurs spontaneously under unfavorable conditions that disrupt the integrity of the folded conformation. Thus, the chemical and physical environment of a protein, such as significant changes in pH...
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Viral Structure00:56

Viral Structure

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Viruses are extraordinarily diverse in shape and size, but they all have several structural features in common. All viruses have a core that contains a DNA- or RNA-based genome. The core is surrounded by a protective coat of proteins called the capsid. The capsid is composed of subunits called capsomeres. The capsid and genome-containing core are together known as the nucleocapsid.
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Alternative In Vitro Methods for the Determination of Viral Capsid Structural Integrity
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Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure.

Emma K Loveday1, Kyle S Hain2, Irina Kochetkova2

  • 1Department of Chemical & Biological Engineering, Montana State University, Bozeman, MT 59717, USA.

Viruses
|April 3, 2021
PubMed
Summary

Two methods, heat treatment and UV irradiation, were validated for inactivating SARS-CoV-2. UV irradiation preserved viral RNA better than heat, impacting downstream assays for Severe Acute Respiratory Syndrome Coronavirus -2 research.

Keywords:
ELISASARS-CoV-2UV inactivationelectron microscopyheat inactivationwestern blot

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Area of Science:

  • Virology
  • Biosafety
  • Molecular Biology

Background:

  • Research on Severe Acute Respiratory Syndrome Coronavirus -2 (SARS-CoV-2) typically requires a biosafety level 3 (BSL3) facility.
  • Inactivating SARS-CoV-2 allows for research at lower biosafety levels, expediting scientific discovery.
  • Effective inactivation protocols are crucial to ensure safety while preserving viral integrity for downstream applications.

Purpose of the Study:

  • To validate and compare heat treatment and ultraviolet (UV) irradiation for SARS-CoV-2 inactivation.
  • To optimize inactivation methods to ensure complete loss of infectivity while minimizing damage to viral components.
  • To assess the impact of each inactivation method on viral RNA, protein yield, and virion quality for various research assays.

Main Methods:

  • SARS-CoV-2 inactivation using optimized heat treatment (65 °C for 15 min) and UV irradiation (70,000 µJ/cm²).
  • Validation of inactivation efficacy through infectivity assays.
  • Comparison of methods using viral RNA quantification (RT-qPCR), virion purification, and protein analysis.

Main Results:

  • Both 15 min of 65 °C heat treatment and 70,000 µJ/cm² UV irradiation achieved complete SARS-CoV-2 inactivation.
  • UV irradiation caused a 2-log reduction in detectable viral RNA genomes compared to heat inactivation.
  • Protein yield was comparable, but viral protein and virion quality varied depending on the inactivation method and duration.

Conclusions:

  • Both heat and UV methods effectively inactivate SARS-CoV-2, enabling safer research.
  • UV irradiation is preferable for assays requiring intact viral RNA, while heat treatment may be suitable when RNA integrity is less critical.
  • Choosing the appropriate inactivation method depends on specific research objectives and downstream assay requirements.