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@ARTICLE{Soh:600224,
author = {Soh, Timothy K. and Pfefferle, Susanne and Wurr, Stephanie
and von Possel, Ronald and Oestereich, Lisa and Rieger, Toni
and Uetrecht, Charlotte and Rosenthal, Maria and Bosse, Jens
Bernhard},
title = {{A} validated protocol to {UV}-inactivate {SARS}-{C}o{V}-2
and herpesvirus-infected cells},
journal = {PLOS ONE},
volume = {18},
number = {5},
issn = {1932-6203},
address = {San Francisco, California, US},
publisher = {PLOS},
reportid = {PUBDB-2023-07798},
pages = {e0274065 -},
year = {2023},
abstract = {Downstream analysis of virus-infected cell samples, such as
reverse transcription polymerase chain reaction (RT PCR) or
mass spectrometry, often needs to be performed at lower
biosafety levels than their actual cultivation, and thus the
samples require inactivation before they can be transferred.
Common inactivation methods involve chemical crosslinking
with formaldehyde or denaturing samples with strong
detergents, such as sodium dodecyl sulfate. However, these
protocols destroy the protein quaternary structure and
prevent the analysis of protein complexes, albeit through
different chemical mechanisms. This often leads to studies
being performed in over-expression or surrogate model
systems. To address this problem, we generated a protocol
that achieves the inactivation of infected cells through
ultraviolet (UV) irradiation. UV irradiation damages viral
genomes and crosslinks nucleic acids to proteins but leaves
the overall structure of protein complexes mostly intact.
Protein analysis can then be performed from intact cells
without biosafety containment. While UV treatment protocols
have been established to inactivate viral solutions, a
protocol was missing to inactivate crude infected cell
lysates, which heavily absorb light. In this work, we
develop and validate a UV inactivation protocol for
SARS-CoV-2, HSV-1, and HCMV-infected cells. A fluence of
10,000 mJ/cm2 with intermittent mixing was sufficient to
completely inactivate infected cells, as demonstrated by the
absence of viral replication even after three sequential
passages of cells inoculated with the treated material. The
herein described protocol should serve as a reference for
inactivating cells infected with these or similar viruses
and allow for the analysis of protein quaternary structure
from bona fide infected cells.},
cin = {CSSB-MHH-JB / CSSB-LIV/DESY-CU / FS-CS},
ddc = {610},
cid = {I:(DE-H253)CSSB-MHH-JB-20210520 /
$I:(DE-H253)CSSB-LIV_DESY-CU-20220525$ /
I:(DE-H253)FS-CS-20210408},
pnm = {633 - Life Sciences – Building Blocks of Life: Structure
and Function (POF4-633)},
pid = {G:(DE-HGF)POF4-633},
experiment = {EXP:(DE-MLZ)NOSPEC-20140101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:37163509},
UT = {WOS:001024735700001},
doi = {10.1371/journal.pone.0274065},
url = {https://bib-pubdb1.desy.de/record/600224},
}