title: Impact of recombinant Drosophila S2 cell population enrichment on expression of rabies virus glycoprotein.
abstract: 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.
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil.
authors.forename: Nayara G L
authors.initials: NG
authors.lastname: Santos
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil.
authors.forename: Mayra P
authors.initials: MP
authors.lastname: Rocca
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Pereira
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil.
authors.forename: Daniella C
authors.initials: DC
authors.lastname: Ventini
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil.
authors.forename: Ana Lia P
authors.initials: AL
authors.lastname: Puglia
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil.
authors.forename: Soraia A C
authors.initials: SA
authors.lastname: Jorge
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil.
authors.forename: Marcos A N
authors.initials: MA
authors.lastname: Lemos
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil. renato.astray@butantan.gov.br.
authors.forename: Renato M
authors.initials: RM
authors.lastname: Astray
keywords: Cycloheximide; Immunomagnetic selection; RT-qPCR; Rabies virus glycoprotein; S2 cells; mRNA