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.

Multi-step purification strategy for RANTES wild-type and mutated analogues expressed in a baculovirus system / B. Comuzzi, C. Arcelloni, S. Polo, V. Nardese, P. Lusso, R. Paroni. - In: JOURNAL OF CHROMATOGRAPHY B. BIOMEDICAL SCIENCES AND APPLICATIONS. - ISSN 1387-2273. - 737:1-2(2000 Jan), pp. 47-54.

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

S. Polo;R. Paroni
2000

Abstract

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.
Baculovirus; Chemokines; Purification; RANTES mutagenesis
Settore BIO/10 - Biochimica
Settore MED/04 - Patologia Generale
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/19752
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