Optimization of the production of full‐length rCD4 in baculovirus‐infected Sf9 cells

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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 × 106 cells/mL used to multiply cells at a maximum exponential growth of 4.5 × 106; (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.