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Optimization of the production of full‐length rCD4 in baculovirus‐infected Sf9 cells
Author(s) -
Lazarte Jaime E.,
Tosi PierreFrancois,
Nicolau Claude
Publication year - 1992
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260400204
Subject(s) - sf9 , production (economics) , baculoviridae , biology , recombinant dna , spodoptera , genetics , economics , gene , macroeconomics
The baculovirus‐insect cell system is reliable in expressing a variety of recombinant proteins. A recombinant baculovirus encoding the full length human CD4 has been used to infect Spodoptera frugiperda 9 cells in 6‐L‐airlift fermentors. The procedured described in this report permitted a 6.5‐fold enhancement of rCD4 expression as compared to standard procedures previously published. The increase of rCD4 expression on the cell surface was achieved by using the following steps: (1) Optimal seeding density of 0.8 × 10 6 cells/mL used to multiply cells at a maximum exponential growth of 4.5 × 10 6 ; (2) high multiplicity of infection (MOI) of 580 PFU/cell; (3) addition of medium at time of infection. In addition to full‐length rCD4, a “short” rCD4 with largely deleted cytoplasmic sequence (last 31 C‐terminal amino acids) was also efficiently expressed.