Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form

Optimization of environmental factors for the production and handling of recombinant retrovirus

Academic Article
uri icon
  • Overview
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

authors

  • Lee, S. G.
  • Kim, S.
  • Robbins, Paul D.
  • Kim, B. G.

publication date

  • May 1996

journal

  • Applied Microbiology and Biotechnology  Journal

abstract

  • Certain steps from the production to infection of the amphotropic retroviral vector, MFG- LacZ, were optimized and the factors that affect retroviral titers were analyzed. Retroviral vector titers were highest when the culture supernatant was harvested 3 days after the producer cells had reached confluence. About a 2-fold increase in vector production was achieved at 32 degrees C compared to that at 37 degrees C. Low serum concentrations had no significant effect on the titers of virus produced by the CRIP cell line. Retroviral vectors were stable at 4 degrees C but very unstable at 37 degrees C and were quite sensitive to freezing and thawing. About 30%-50% of viral infectivity was lost during the thawing step and the loss was not recovered by the addition of commonly used cryoprotectants. Increase in viral exposure time for infection to target NIH3T3 cells was linearly proportional to the retroviral titer for up to 15 h. In addition, using DEAE-dextran in place of polybrene as a polycation during infection enhanced infection efficiency about 3-fold. The retrovirus was robust to simple ultrafiltration and its titer could be easily concentrated 16-fold. Taken together, our data suggest that at least a 100-fold increase in titer can be achieved with simple optimization.

subject areas

  • 3T3 Cells
  • Animals
  • Blood Physiological Phenomena
  • Cattle
  • Culture Media
  • Genetic Vectors
  • Mice
  • Retroviridae
  • Temperature
  • Ultrafiltration
  • Virus Cultivation
  • Virus Replication
scroll to property group menus

Identity

International Standard Serial Number (ISSN)

  • 0175-7598

PubMed ID

  • 8737571
scroll to property group menus

Additional Document Info

start page

  • 477

end page

  • 483

volume

  • 45

issue

  • 4

©2021 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support