title: Optimization of the production of full-length rCD4 in baculovirus-infected Sf9 cells.
abstract: 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 x 10(6) cells/mL used to multiply cells at a maximum exponential growth of 4.5 x 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.
authors.affiliation: Cell Biology Section, Institute of BioSciences and Technology, Texas A&M University, College Station, Texas 77843, USA.
authors.forename: J E
authors.initials: JE
authors.lastname: Lazarte
authors.forename: P F
authors.initials: PF
authors.lastname: Tosi
authors.forename: C
authors.initials: C
authors.lastname: Nicolau