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Improved microscale cultivation of Pichia pastoris for clonal screening

Overview of attention for article published in Fungal Biology and Biotechnology, May 2018
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2 tweeters

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Title
Improved microscale cultivation of Pichia pastoris for clonal screening
Published in
Fungal Biology and Biotechnology, May 2018
DOI 10.1186/s40694-018-0053-6
Pubmed ID
Authors

Alexander Eck, Matthias Schmidt, Stefanie Hamer, Anna Joelle Ruff, Jan Förster, Ulrich Schwaneberg, Lars M. Blank, Wolfgang Wiechert, Marco Oldiges

Abstract

Expanding the application of technical enzymes, e.g., in industry and agriculture, commands the acceleration and cost-reduction of bioprocess development. Microplates and shake flasks are massively employed during screenings and early phases of bioprocess development, although major drawbacks such as low oxygen transfer rates are well documented. In recent years, miniaturization and parallelization of stirred and shaken bioreactor concepts have led to the development of novel microbioreactor concepts. They combine high cultivation throughput with reproducibility and scalability, and represent promising tools for bioprocess development. Parallelized microplate cultivation of the eukaryotic protein production host Pichia pastoris was applied effectively to support miniaturized phenotyping of clonal libraries in batch as well as fed-batch mode. By tailoring a chemically defined growth medium, we show that growth conditions are scalable from microliter to 0.8 L lab-scale bioreactor batch cultivation with different carbon sources. Thus, the set-up allows for a rapid physiological comparison and preselection of promising clones based on online data and simple offline analytics. This is exemplified by screening a clonal library of P. pastoris constitutively expressing AppA phytase from Escherichia coli. The protocol was further modified to establish carbon-limited conditions by employing enzymatic substrate-release to achieve screening conditions relevant for later protein production processes in fed-batch mode. The comparison of clonal rankings under batch and fed-batch-like conditions emphasizes the necessity to perform screenings under process-relevant conditions. Increased biomass and product concentrations achieved after fed-batch microscale cultivation facilitates the selection of top producers. By reducing the demand to conduct laborious and cost-intensive lab-scale bioreactor cultivations during process development, this study will contribute to an accelerated development of protein production processes.

Twitter Demographics

The data shown below were collected from the profiles of 2 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 41 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 41 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 12 29%
Student > Ph. D. Student 7 17%
Student > Master 6 15%
Student > Bachelor 3 7%
Professor 1 2%
Other 3 7%
Unknown 9 22%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 13 32%
Agricultural and Biological Sciences 10 24%
Chemical Engineering 2 5%
Immunology and Microbiology 2 5%
Engineering 2 5%
Other 1 2%
Unknown 11 27%

Attention Score in Context

This research output has an Altmetric Attention Score of 2. 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 04 May 2018.
All research outputs
#7,691,268
of 12,889,535 outputs
Outputs from Fungal Biology and Biotechnology
#46
of 61 outputs
Outputs of similar age
#147,050
of 269,062 outputs
Outputs of similar age from Fungal Biology and Biotechnology
#3
of 4 outputs
Altmetric has tracked 12,889,535 research outputs across all sources so far. This one is in the 39th percentile – i.e., 39% of other outputs scored the same or lower than it.
So far Altmetric has tracked 61 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 5.4. This one is in the 18th percentile – i.e., 18% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 269,062 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 44th percentile – i.e., 44% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 4 others from the same source and published within six weeks on either side of this one.