Multi-step purification strategy for RANTES wild-type and mutated analogues expressed in a baculovirus system

RANTES (regulated on activation, normal T cell expressed and secreted), a C-C chemokine, is one of the major HIV-suppressive factors produced by CD8+ T cells. Wild-type RANTES and genetically modified analogues were expressed in a baculovirus system and purified from cell culture supernatants employing a multi-step strategy based on affinity and RP-HPLC. Quantification and purity control of the final proteins were carried out by capillary electrophoresis using the synthetic or the recombinant wild-type RANTES as a reference. The procedure here reported requires only three days to obtain 0.016–0.270 mg of the pure and characterised proteins, starting from 370–900 ml of culture media, and is suitable for the analysis of a large number of RANTES analogues