Behavior of Wild-type and Transfected S2 Cells Cultured
in Two Different Media
Fabiana R. X. Batista
&
Kátia N. Greco
&
Renato M. Astray
&
Soraia A. C. Jorge
&
Elisabeth F. P. Augusto
&
Carlos A. Pereira
&
Ronaldo Z. Mendonça
&
Ângela M. Moraes
Received: 29 October 2009 /Accepted: 17 January 2010 /
Published online: 15 December 2010
#
Springer Science+Business Media, LLC 2010
Abstract An animal protein-free medium composed of IPL-41 containing 6 g L
−1
yeastolate ultrafiltrate, 10 g L
−1
glucose, 2 g L
−1
lactose, 5 g L
−1
glutamine, 1% lipid
emulsion, and 0.1% Pluronic F-68 was used for producing recombinant proteins in batch
mode employing two cell lines, S2AcRVGP2k expressing the G glycoprotein from rabies
virus (RVGP) and S2AcHBsAgHy-9C expressing the surface antigen of hepatitis B virus
(HBsAg), both obtained from Drosophila melanogaster S2 cells. Growth of wild-type S2
cells was also evaluated in the same medium. Cell behavior in the tested medium was
compared to that verified in Sf900 II®. The results show that in shake flasks, S2AcRVGP2k
and S2AcHBsAgHy-9C cells reached around 2×10
7
cells mL
−1
in both media. In
supplemented IPL-41 and Sf900 II® media, S2AcRVGP2k cells produced
367 ng RVGP mL
−1
and 638 ng RVGP mL
−1
, respectively, while S2AcHBsAgHy-9C
cells correspondently produced 573 ng HBsAg mL
−1
and 322 ng HBsAg mL
−1
in the
mentioned media. In stirred tanks, S2AcRVGP2k cells reached 3×10
7
cells mL
−1
and
produced up to 758 ng RVGP mL
−1
. In general, glucose was consumed by cells, while
lactate and ammonia were produced.
Keywords S2 cells
.
Metabolism
.
RVGP
.
HBsAg
.
Animal protein-free medium
Appl Biochem Biotechnol (2011) 163:1–13
DOI 10.1007/s12010-010-8918-z
F. R. X. Batista
:
Â. M. Moraes (*)
Departamento de Processos Biotecnológicos, Faculdade de Engenharia Química, Universidade Estadual
de Campinas, Av. Albert Einstein, 500, 13083-852, Campinas, SP, Brazil
e-mail: ammoraes@feq.unicamp.br
K. N. Greco
:
R. M. Astray
:
S. A. C. Jorge
:
C. A. Pereira
:
R. Z. Mendonça
Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brazil 1500, 05503-900, São Paulo-SP,
Brazil
E. F. P. Augusto
Laboratório de Biotecnologia Industrial, Instituto de Pesquisas Tecnológicas do Estado de São Paulo,
Av Prof. Almeida Prado 532, Prédio 31, 05508-901, São Paulo-SP, Brazil