Tello, DanielRodríguez-Rodríguez, MarYélamos, BelénGómez-Gutiérrez, JuliánPeterson, Darrell L.Gavilanes, Francisco2023-06-182023-06-182015-03-010166-0934; 1879-098410.1016/j.jviromet.2014.11.020https://hdl.handle.net/20.500.14352/24213In this report it is described for the first time the expression and purification of large quantities of a oluble and correctly folded chimeric recombinant protein, E2661E1340, containing the permuted Hepatitis C virus (HCV) glycoprotein ectodomains E1 (amino acids 192-340) and E2 (amino acids 384-661). Using the baculovirus/insect cell expression system, 8mg of secreted protein were purified from 1L of culture media, a yield 4 times higher than the described for its counterpart E1341E2661. This permuted chimeric protein is glycosylated and possesses a high tendency to self-associate. The fluorescence emission spectrum indicates that Trp residues occupy a relatively low hydrophobic environment. The secondary structure was determined by deconvolution of the far-UV circular dichroism spectrum yielding 13% α-helix structure, 49% extended structure and 38% non-ordered structure. E2661E1340 binds to antibodies present in human sera from HCV-positive patients, a binding that is blocked at different levels by a rabbit anti-E2661 antibody. All these structural and antigenic features of E2661E1340 are very similar to those described for E1340E2661, Thus, this high-yield isolated chimeric protein may be a valuable tool to study the first steps of the HCV infection.engjournal articlehttp://www.sciencedirect.com/science/article/pii/S0166093414004509open access577.1Hepatitis C Virusenvelope proteinE1E2baculovirusglycosylationBioquímica (Química)