%0 Journal Article
%A Tello, Daniel
%A Rodríguez-Rodríguez, Mar
%A Yélamos, Belén
%A Gómez-Gutiérrez, Julián
%A Peterson, Darrell L.
%A Gavilanes, Francisco
%T High-yield production of a chimeric glycoprotein based on permuted E1 and E2 HCV envelope ectodomains
%D 2015
%@ 0166-0934; 1879-0984
%U https://hdl.handle.net/20.500.14352/24213
%X In 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.
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