Impact of recombinant Drosophila S2 cell population enrichment on expression of rabies virus glycoprotein

Recombinant Drosophila S2 cells have been used for the expression of many proteins of medical interest. However, membrane-attached glycoproteins, which commonly exhibit lower expression levels compared to soluble proteins, may require special procedures in order to attain high levels of expression....

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Bibliographic Details
Published in:Cytotechnology (Dordrecht) 2016-12, Vol.68 (6), p.2605-2611
Main Authors: Santos, Nayara G. L., Rocca, Mayra P., Pereira, Carlos A., Ventini, Daniella C., Puglia, Ana Lia P., Jorge, Soraia A. C., Lemos, Marcos A. N., Astray, Renato M.
Format: Article
Language:English
Subjects:
Antibodies
Bats
Biochemistry
Biomedicine
Biotechnology
Cell culture
Chemistry
Chemistry and Materials Science
Copy number
Cycloheximide
Drosophila
Glycoproteins
Insects
Lyssavirus
Membranes
mRNA
Original
Original Article
Population studies
Proteins
Rabies
Transformed cells
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Summary:Recombinant Drosophila S2 cells have been used for the expression of many proteins of medical interest. However, membrane-attached glycoproteins, which commonly exhibit lower expression levels compared to soluble proteins, may require special procedures in order to attain high levels of expression. In this study, two S2 cell population enrichment methods (antibiotic and immunomagnetic selection) were evaluated for their ability to enhance expression of the membrane-anchored rabies virus glycoprotein (RVGP). Quantification of RVGP production and determination of its cDNA copy number in transformed cells showed that both enrichment methods increased RVGP expression without significantly affecting its gene copy number. More interestingly, RVGP mRNA levels measured after cycloheximide treatment were poorly correlated with glycoprotein levels. Both enrichment methods enhanced expression of RVGP by recombinant S2 cells, with the highest level of expression achieved using immunomagnetic selection.
ISSN:0920-9069
1573-0778
DOI:10.1007/s10616-016-9984-z