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Cytotechnology

, Volume 17, Issue 1, pp 21–26 | Cite as

Strong buffering capacity of insect cells. Implications for the baculovirus expression system

  • Miguel Medina
  • Abelardo López-Rivas
  • Douwe Zuidema
  • Graham J. Belsham
  • Esteban Domingo
  • Just M. Vlak
Article

Abstract

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 theSpodoptera 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.

Key words

Insect cells intracellular pH baculovirus foot-and-mouth disease virus empty capsids 

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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Miguel Medina
    • 1
  • Abelardo López-Rivas
    • 2
  • Douwe Zuidema
    • 3
  • Graham J. Belsham
    • 4
  • Esteban Domingo
    • 1
  • Just M. Vlak
    • 3
  1. 1.Centro de Biología Molecular ‘Severo Ochoa’CSICMadridSpain
  2. 2.Institute de Parasitología López-NeyraCSICGranadaSpain
  3. 3.Department of VirologyAgricultural UniversityWageningenThe Netherlands
  4. 4.Pirbright LaboratoryAFRC Institute for Animal HealthPirbrightUK

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