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.
|Titolo:||Multi-step purification strategy for RANTES wild-type and mutated analogues expressed in a baculovirus system|
|Parole Chiave:||Baculovirus; Chemokines; Purification; RANTES mutagenesis|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
Settore MED/04 - Patologia Generale
|Data di pubblicazione:||gen-2000|
|Digital Object Identifier (DOI):||10.1016/S0378-4347(99)00361-8|
|Appare nelle tipologie:||01 - Articolo su periodico|