Strong buffering capacity of insect cells. Implications for the baculovirus expression system
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Strong buffering capacity of insect cells. Implications for the baculovirus expression system. / Medina, Miguel; López-Rivas, Abelardo; Zuidema, Douwe; Belsham, Graham J.; Domingo, Esteban; Vlak, Just M.
I: Cytotechnology, Bind 17, Nr. 1, 02.1995, s. 21-26.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - Strong buffering capacity of insect cells. Implications for the baculovirus expression system
AU - Medina, Miguel
AU - López-Rivas, Abelardo
AU - Zuidema, Douwe
AU - Belsham, Graham J.
AU - Domingo, Esteban
AU - Vlak, Just M.
PY - 1995/2
Y1 - 1995/2
N2 - Insect cells are widely used for expression of a variety of different proteins by using the baculovirus expression system. The applicability of this system depends on production of proteins which have biological properties similar to their native counterparts. One application has been the expression of viral capsid proteins and their assembly into empty capsid structures to provide new viral immunogens which retain complex antigenic sites. An important parameter for efficient folding and assembly of proteins into viral procapsids may be the intracellular pH, particularly for acid-labile particles such as foot-and-mouth disease virus (FMDV). Benzoic acid was used as an effective indicator of intracellular pH in insect cells and 3-O-methyl glucose to measure cell volumes. We have determined the intracellular volume of the Spodoptera frugiperda IPLB-Sf21 insect cells 0.50±0.08 pL per cell. Using the distribution of [14C]-benzoic acid, we show that the intracellular pH remains constant at pH 7.0 when the cells are grown in media with pH values ranging from 6.2 to 6.8 and, moreover, is not affected by baculovirus infection. These results suggest that insect cells are suitable to express and produce acid-labile structures via the baculovirus expression system and that assembly of proteins and viral procapsids could occur.
AB - Insect cells are widely used for expression of a variety of different proteins by using the baculovirus expression system. The applicability of this system depends on production of proteins which have biological properties similar to their native counterparts. One application has been the expression of viral capsid proteins and their assembly into empty capsid structures to provide new viral immunogens which retain complex antigenic sites. An important parameter for efficient folding and assembly of proteins into viral procapsids may be the intracellular pH, particularly for acid-labile particles such as foot-and-mouth disease virus (FMDV). Benzoic acid was used as an effective indicator of intracellular pH in insect cells and 3-O-methyl glucose to measure cell volumes. We have determined the intracellular volume of the Spodoptera frugiperda IPLB-Sf21 insect cells 0.50±0.08 pL per cell. Using the distribution of [14C]-benzoic acid, we show that the intracellular pH remains constant at pH 7.0 when the cells are grown in media with pH values ranging from 6.2 to 6.8 and, moreover, is not affected by baculovirus infection. These results suggest that insect cells are suitable to express and produce acid-labile structures via the baculovirus expression system and that assembly of proteins and viral procapsids could occur.
KW - baculovirus
KW - empty capsids
KW - foot-and-mouth disease virus
KW - Insect cells
KW - intracellular pH
UR - http://www.scopus.com/inward/record.url?scp=0028873693&partnerID=8YFLogxK
U2 - 10.1007/BF00749217
DO - 10.1007/BF00749217
M3 - Journal article
AN - SCOPUS:0028873693
VL - 17
SP - 21
EP - 26
JO - Cytotechnology
JF - Cytotechnology
SN - 0920-9069
IS - 1
ER -
ID: 379029544