RT Journal Article
T1 High-yield production of a chimeric glycoprotein based on permuted E1 and E2 HCV envelope ectodomains
A1 Tello, Daniel
A1 Rodríguez-Rodríguez, Mar
A1 Yélamos, Belén
A1 Gómez-Gutiérrez, Julián
A1 Peterson, Darrell L.
A1 Gavilanes, Francisco
AB 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.
PB Elsevier Science BV
SN 0166-0934; 1879-0984
YR 2015
FD 2015-03-01
LK https://hdl.handle.net/20.500.14352/24213
UL https://hdl.handle.net/20.500.14352/24213
LA eng
DS Docta Complutense
RD 10 abr 2025