| Title |
Multiple reaction monitoring targeted LC-MS analysis of potential cell death marker proteins for increased bioprocess control
|
|---|---|
| Published in |
Analytical & Bioanalytical Chemistry, April 2018
|
| DOI | 10.1007/s00216-018-1029-3 |
| Pubmed ID | |
| Authors |
Simone Albrecht, Christian Kaisermayer, David Reinhart, Monica Ambrose, Renate Kunert, Anna Lindeberg, Jonathan Bones |
| Abstract |
The monitoring of protein biomarkers for the early prediction of cell stress and death is a valuable tool for process characterization and efficient biomanufacturing control. A representative set of six proteins, namely GPDH, PRDX1, LGALS1, CFL1, TAGLN2 and MDH, which were identified in a previous CHO-K1 cell death model using discovery LC-MSEwas translated into a targeted liquid chromatography multiple reaction monitoring mass spectrometry (LC-MRM-MS) platform and verified. The universality of the markers was confirmed in a cell growth model for which three Chinese hamster ovary host cell lines (CHO-K1, CHO-S, CHO-DG44) were grown in batch culture in two different types of basal media. LC-MRM-MS was also applied to spent media (n = 39) from four perfusion biomanufacturing series. Stable isotope-labelled peptide analogues and a stable isotope-labelled monoclonal antibody were used for improved protein quantitation and simultaneous monitoring of the workflow reproducibility. Significant increases in protein concentrations were observed for all viability marker proteins upon increased dead cell numbers and allowed for discrimination of spent media with dead cell densities below and above 1 × 106 dead cells/mL which highlights the potential of the selected viability marker proteins in bioprocess control. Graphical abstract Overview of the LC-MRM-MS workflow for the determination of proteomic markers in conditioned media from the bioreactor that correlate with CHO cell death. |
X Demographics
The data shown below were collected from the profiles of 8 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Russia | 1 | 13% |
| Germany | 1 | 13% |
| United States | 1 | 13% |
| Unknown | 5 | 63% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 4 | 50% |
| Scientists | 2 | 25% |
| Practitioners (doctors, other healthcare professionals) | 1 | 13% |
| Science communicators (journalists, bloggers, editors) | 1 | 13% |
Mendeley readers
The data shown below were compiled from readership statistics for 29 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 29 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 5 | 17% |
| Student > Ph. D. Student | 4 | 14% |
| Other | 2 | 7% |
| Student > Bachelor | 2 | 7% |
| Professor | 1 | 3% |
| Other | 3 | 10% |
| Unknown | 12 | 41% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 5 | 17% |
| Chemical Engineering | 3 | 10% |
| Chemistry | 3 | 10% |
| Agricultural and Biological Sciences | 2 | 7% |
| Unspecified | 1 | 3% |
| Other | 2 | 7% |
| Unknown | 13 | 45% |
Attention Score in Context
This research output has an Altmetric Attention Score of 12. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 08 January 2020.
All research outputs
#3,027,614
of 25,382,440 outputs
Outputs from Analytical & Bioanalytical Chemistry
#284
of 9,619 outputs
Outputs of similar age
#60,529
of 342,742 outputs
Outputs of similar age from Analytical & Bioanalytical Chemistry
#8
of 189 outputs
Altmetric has tracked 25,382,440 research outputs across all sources so far. Compared to these this one has done well and is in the 88th percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 9,619 research outputs from this source. They receive a mean Attention Score of 3.1. This one has done particularly well, scoring higher than 97% of its peers.
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We're also able to compare this research output to 189 others from the same source and published within six weeks on either side of this one. This one has done particularly well, scoring higher than 95% of its contemporaries.