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Secretion of Fc-amidated peptide fusion proteins by Chinese hamster ovary cells

Overview of attention for article published in BMC Biotechnology, June 2015
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  • Good Attention Score compared to outputs of the same age (73rd percentile)

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Title
Secretion of Fc-amidated peptide fusion proteins by Chinese hamster ovary cells
Published in
BMC Biotechnology, June 2015
DOI 10.1186/s12896-015-0173-5
Pubmed ID
Authors

Kristina R. Carlson, Steven C. Pomerantz, Jiali Li, Omid Vafa, Michael Naso, William Strohl, Richard E. Mains, Betty A. Eipper

Abstract

The therapeutic use of α-amidated peptides (e.g. calcitonin, glucagon-like peptide) has increased dramatically, but there are major impediments to wider use of such peptides. Larger peptides are expensive to synthesize, and short plasma half-lives frequently limit the clinical circumstances in which the peptides would be useful. Both problems are potentially solved by producing peptides as fusions with the Fc region of human immunoglobulin. Glucagon-like peptide 1 (GLP1), peptide YY (PYY) and neuromedin U (NMU) were expressed and purified from stable CHO lines; since the α-amide group is essential for full biological potency of many peptides, Fc-fusion peptides were expressed in CHO lines stably expressing the α-amidating enzyme, peptidylglycine α-amidating monooxygenase (PAM: EC 1.14.17.3). Purified fusion proteins were analyzed intact and after HRV3C rhinovirus protease cleavage, at a site in the linker separating the Fc region from the peptide, by mass spectrometry and amide-specific immunoassays. The Fc fusions were expressed at 1-2.5 μg/mg cell protein and secreted at 5-20 % of cell content per hour, in a peptide-specific manner. CHO cells express measurable endogenous PAM activity, amidating 25 % of Fc-PYY and almost 90 % of Fc-GLP1. Expression of exogenous PAM increased the level of peptide amidation to 50 % of Fc-PYY and 95 % of Fc-NMU. The Fc-GLP1 fusions were 10,000-fold less active than synthetic GLP1 in a cell-receptor cyclic AMP-based assay, as expected since the amino terminal of GLP1 is essential for full biological activity. The Fc-PYY fusions were 100-fold less active than PYY-NH2 but 10-fold more active than non-amidated PYY-Gly. This type of approach can be used for the production of stabilized α-amidated peptides aimed at clinical trials.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 8 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 8 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 3 38%
Professor > Associate Professor 1 13%
Other 1 13%
Unknown 3 38%
Readers by discipline Count As %
Agricultural and Biological Sciences 2 25%
Biochemistry, Genetics and Molecular Biology 2 25%
Pharmacology, Toxicology and Pharmaceutical Science 1 13%
Unknown 3 38%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 5. 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 25 August 2022.
All research outputs
#6,153,850
of 23,164,913 outputs
Outputs from BMC Biotechnology
#346
of 941 outputs
Outputs of similar age
#69,836
of 263,955 outputs
Outputs of similar age from BMC Biotechnology
#30
of 39 outputs
Altmetric has tracked 23,164,913 research outputs across all sources so far. This one has received more attention than most of these and is in the 73rd percentile.
So far Altmetric has tracked 941 research outputs from this source. They typically receive more attention than average, with a mean Attention Score of 7.8. This one has gotten more attention than average, scoring higher than 63% of its peers.
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 263,955 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 73% of its contemporaries.
We're also able to compare this research output to 39 others from the same source and published within six weeks on either side of this one. This one is in the 23rd percentile – i.e., 23% of its contemporaries scored the same or lower than it.