Cell-Engineered Recombinant α-Synuclein: A Gage R&R Validated Protocol
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14110%2F24%3A00139081" target="_blank" >RIV/00216224:14110/24:00139081 - isvavai.cz</a>
Result on the web
<a href="https://pubs.acs.org/doi/10.1021/acs.jproteome.3c00190" target="_blank" >https://pubs.acs.org/doi/10.1021/acs.jproteome.3c00190</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1021/acs.jproteome.3c00190" target="_blank" >10.1021/acs.jproteome.3c00190</a>
Alternative languages
Result language
angličtina
Original language name
Cell-Engineered Recombinant α-Synuclein: A Gage R&R Validated Protocol
Original language description
alpha-Synuclein (alpha-Syn) misfolding and its presence in Lewy bodies are observed in almost all Parkinson's disease (PD) patients. Basic biomedical research would benefit from a quick, low-cost approach to purifying alpha-Syn and developing in vitro and in vivo models for PD. Several research groups utilize PFF-based models, yet the production of alpha-Syn PFFs is inconsistent, resulting in nonconclusive findings. Some research laboratories prepare recombinant alpha-Syn (r alpha-Syn) by molecular cloning to overexpress alpha-Syn with various purifying techniques. Laboratory-to-laboratory protocols cause considerable variability and sometimes contradictory findings. PD researchers spend more on protein than solving alpha-Syn's riddles. This article uncovered a novel method for expressing and purifying r alpha-Syn validated through gage reproducibility and repeatability (Gage R&R). For the production of r alpha-Syn, we have employed the ability of a high-cell-density-based expression system to overexpress protein in BL21(DE3). A simple, high-throughput, nonchromatographical purification protocol has been devised to facilitate research with higher reproducibility, which was validated through Gage R&R. A crossover experimental design was utilized, and the purified protein was characterized using orthogonal high-end analytical methods, which displayed higher similarity between the isolated r alpha-Syn. Batch-to-batch variability was the least for produced protein and hence can be utilized for exploring the iceberg of PD.
Czech name
—
Czech description
—
Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
—
OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
—
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2024
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Proteome Research
ISSN
1535-3893
e-ISSN
1535-3907
Volume of the periodical
23
Issue of the periodical within the volume
1
Country of publishing house
US - UNITED STATES
Number of pages
9
Pages from-to
16-24
UT code for WoS article
001137570700001
EID of the result in the Scopus database
2-s2.0-85179151181