About: Serum‐free transient protein production system based on adenoviral vector and PER.C6 technology: High yield and preserved bioactivity

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About: Serum‐free transient protein production system based on adenoviral vector and PER.C6 technology: High yield and preserved bioactivity Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Serum‐free transient protein production system based on adenoviral vector and PER.C6 technology: High yield and preserved bioactivity
Creator	Smits, S Goudsmit, J Brouwer, K Hateboer, G Havenga, M Heemskerk, E Holterman, L Kaspers, J Koldijk, M Melis, I Schouten, G Van Der Vlugt, R Vogels, R
Source	Medline; PMC
abstract	Stable E1 transformed cells, like PER.C6, are able to grow at scale and to high cell densities. E1‐deleted adenoviruses replicate to high titer in PER.C6 cells whereas subsequent deletion of E2A from the vector results in absence of replication in PER.C6 cells and drastically lowers the expression of adenovirus proteins in such cells. We therefore considered the use of an ΔE1/ΔE2 type 5 vector (Ad5) to deliver genes to PER.C6 cells growing in suspension with the aim to achieve high protein yield. To evaluate the utility of this system we constructed ΔE1/ΔE2 vector carrying different classes of protein, that is, the gene coding for spike protein derived from the Coronavirus causing severe acute respiratory syndrome (SARS‐CoV), a gene coding for the SARS‐CoV receptor or the genes coding for an antibody shown to bind and neutralize SARS‐CoV (SARS‐AB). The ΔE1/ΔE2A‐vector backbones were rescued on a PER.C6 cell line engineered to constitutively over express the Ad5 E2A protein. Exposure of PER.C6 cells to low amounts (30 vp/cell) of ΔE1/ΔE2 vectors resulted in highly efficient (>80%) transduction of PER.C6 cells growing in suspension. The efficient cell transduction resulted in high protein yield (up to 60 picogram/cell/day) in a 4 day batch production protocol. FACS and ELISA assays demonstrated the biological activity of the transiently produced proteins. We therefore conclude that ΔE1/ΔE2 vectors in combination with the PER.C6 technology may provide a viable answer to the increasing demand for high quality, high yield recombinant protein. Biotechnol. Bioeng. 2008;100: 273–283. © 2007 Wiley Periodicals, Inc.
has issue date	2007-12-13(xsd:dateTime)
bibo:doi	10.1002/bit.21757
bibo:pmid	18512821
has license	no-cc
sha1sum (hex)	83263b1c3aa6f156aef5edc952a10b1d8e8546e0
schema:url	https://doi.org/10.1002/bit.21757
resource representing a document's title	Serum‐free transient protein production system based on adenoviral vector and PER.C6 technology: High yield and preserved bioactivity
has PubMed Central identifier	PMC7161845
has PubMed identifier	18512821
schema:publication	Biotechnol Bioeng
resource representing a document's body	covid:83263b1c3aa6f156aef5edc952a10b1d8e8546e0#body_text
is schema:about of	named entity 'SUSPENSION' named entity 'BATCH' named entity 'STABLE' named entity 'spike protein' named entity 'vector' named entity 'gene coding' named entity 'severe acute respiratory syndrome' named entity 'DE2' named entity 'Coronavirus' named entity 'titer' named entity 'antibody' named entity 'protein' named entity 'protein' named entity 'Protein Production' named entity 'High Yield' named entity 'SARS' named entity 'transformed cells' named entity 'Adenoviral Vector' named entity 'vector' named entity 'gene coding' named entity 'vector' named entity 'SARS-CoV' named entity 'DE1' named entity 'cell line' named entity 'protein' named entity 'adenovirus' named entity 'DERIVED' named entity 'A GENE' named entity 'SCALE' named entity 'CELL DENSITIES' named entity 'TYPE 5' named entity 'CONSTRUCTED' named entity 'BIOACTIVITY' named entity 'TECHNOLOGY' named entity 'PROTEIN PRODUCTION' named entity 'SARS-CoV' named entity 'vector' named entity 'vectors' named entity 'VIABLE' named entity 'CARRYING' named entity 'SARS-COV' named entity 'EXPRESS' named entity 'AD5' named entity 'TRANSFORMED' named entity 'DEMAND' named entity 'GROWING' named entity 'CORONAVIRUS' named entity 'TRANSDUCTION' named entity 'EFFICIENT' named entity 'BIOLOGICAL ACTIVITY' named entity 'HIGH YIELD' named entity 'TO DELIVER' named entity 'INCREASING' named entity 'REPLICATE' named entity 'C6 CELL' named entity 'Ad5' named entity 'FACS' named entity 'receptor' named entity 'adenoviruses' named entity 'Bioactivity' covid:arg/83263b1c3aa6f156aef5edc952a10b1d8e8546e0 named entity 'VECTOR' named entity 'AMOUNTS' named entity 'ABSENCE OF' named entity 'DIFFERENT' named entity 'E2A' named entity 'CONCLUDE' named entity 'PROTEINS' named entity 'ABLE TO'

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