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Cytotechnology

, Volume 21, Issue 2, pp 155–163 | Cite as

The kinetics of baculovirus adsorption to insect cells in suspension culture

  • John F. Power
  • Steven Reid
  • Paul F. Greenfield
  • Lars K. Nielsen
Special Issue

Abstract

The influence of various culture parameters on the attachment of a recombinant baculovirus to suspended insect cells was examined under normal culture conditions. These parameters included cell density, multiplicity of infection, and composition of the cell growth medium. It was found that the fractional rate of virus attachment was independent of the multiplicity of infection but dependent on the cell density. A first order mathematical model was used to simulate the adsorption kinetics and predict the efficiency of virus attachment under the various culture conditions. This calculated efficiency of virus attachment was observed to decrease at high cell densities, which was attributed to cell clumping. It was also observed that virus attachment was more efficient in Sf900II serum free medium than it was in IPL-41 serum-supplemented medium. This effect was attributed to the protein in serum which may coat the cells and so inhibit adsorption. A general discussion relating the observations made in-these experiments to the kinetics of recombinant baculovirus adsorption to suspended insect cells is presented.

Key words

Autographa californica NPV baculovirus expression vector system insect cell culture Spodoptera frugiperda suspension culture virus adsorption kinetics 

Abbreviations

BEVS

Baculovirus expression vector system

FBS

Foetal bovine serum

MOI

Multiplicity of infection

NPV

Nuclear polyhedrosis virus

PFU

Plaque forming unit

Nomenclature

DB

diffusion coefficient calculated by Stokes-Einstein equation (m2s−1)

f

fraction of the initially suspended virus particles adsorbed

k

Boltzmann's constant (1.38 × 10−23 J K-−1)

R

cell radius (10−5 m)

r

virus particle radius (2 × 10−7 m)

T

absolute temperature (K)

t

adsorption time (hour)

X

cell concentration (106 ml−1)

aA

first order virus binding coefficient (hour−1)

h

medium viscosity (cp)

f

fraction of virus-cell collisions which result in attachment

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Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • John F. Power
    • 1
  • Steven Reid
    • 1
  • Paul F. Greenfield
    • 1
  • Lars K. Nielsen
    • 1
  1. 1.Department of Chemical EngineeringThe University of QueenslandQueenslandAustralia

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