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 express ed in a baculovirus system and purified from cell culture supernatants empl oying a multi-step strategy based on affinity and RP-HPLC. Quantification a nd purity control of the final proteins were carried out by capillary elect rophoresis using the synthetic or the recombinant wild-type RANTES as a ref erence. The procedure here reported requires only three days to obtain 0.01 6-0.270 mg of the pure and characterised proteins, starting from 370-900 mi of culture media, and is suitable for the analysis of a large number of RA NTES analogues (C) 2000 Elsevier Science B.V. All rights reserved.