Heterologous protein expression affects the death kinetics of baculovirus-infected insect cell cultures: a quantitative study by use of n-target theory

Biotechnol Prog. 1994 Jan-Feb;10(1):55-9. doi: 10.1021/bp00025a006.

Abstract

The death of cultured insect cells after baculovirus infection is a time-dependent event. Without a quantitative model, it is difficult to characterize its kinetics. Our group has shown that the cell survival rate can be characterized by use of the n-target theory, which involves only two parameters: the number of hypothetical inactivation targets (n) and the first-order death rate (k). In this study, we used different recombinant viruses to examine the effect of heterologous protein expression on the cell survival rate. The proteins expressed were beta-galactosidase, human T-cell leukemia virus type I p40x, human interleukin-2, and human tissue plasminogen activator (tPA). The survival rate was affected by protein expression, but the n value remained constant if the protein expression level was high (above 30 mg/L). Low-level expression of secreted, glycosylated tPA resulted in a reduced n value, which was restored to the normal value when the tPA signal peptide and prosequence were deleted. In addition, if the n value was normal (10-11), the level of protein expression correlated negatively with the death rate. However, if the n value was reduced by unfavorable culture conditions or foreign protein expression, the expression level correlated positively with the death rate. A dimensionless plot with kt as the dimensionless time shows that alteration of the k value while retaining constant n is equivalent to a rescaling of time. Therefore, the survival curves with constant n reduce to a single curve on the dimensionless plot.(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Cell Death*
  • Cell Survival
  • Cells, Cultured
  • Interleukin-2 / biosynthesis*
  • Moths
  • Occlusion Body Matrix Proteins
  • Promoter Regions, Genetic
  • Recombinant Proteins / biosynthesis*
  • Tissue Plasminogen Activator / biosynthesis*
  • Viral Proteins / genetics
  • Viral Structural Proteins
  • beta-Galactosidase / biosynthesis*

Substances

  • Interleukin-2
  • Occlusion Body Matrix Proteins
  • Recombinant Proteins
  • Viral Proteins
  • Viral Structural Proteins
  • polyhedrin protein, Nucleopolyhedrovirus
  • beta-Galactosidase
  • Tissue Plasminogen Activator