HCV E2 construct compositions and methods

A method of producing a Hepatitis C virus (HCV) eE2 polypeptide comprising:

• (i) providing a construct comprising:
• a cytomegalovirus (CMV) promoter,
• a prolactin signal sequence,
• a nucleic acid sequence encoding an amino acid sequence of an ectodomain of a HCV E2 (HCV eE2) polypeptide,
• a thrombin cleavage site,
• wherein the CMV promoter is followed by the prolactin signal sequence, the prolactin signal sequence is followed by the nucleic acid sequence, the nucleic acid sequence is followed by the thrombin cleavage site, and
• the amino acid sequence is identical to the amino acid sequence consisting of amino acids 384-656 of SEQ ID NO:1 of HCV 2a genotype, isolate J6, or the corresponding sequence of a different HCV genotype or the corresponding sequence of SEQ ID NO:
• 15, SEQ ID NO:
• 16, SEQ ID NO:
• 17, SEQ ID NO:
• 18, SEQ ID NO:
• 19, SEQ ID NO:
• 20, or SEQ ID NO:
• 21, and fragments thereof that are 50 to 270 amino acids in length,
• (ii) introducing the construct into mammalian cells
• (iii) selecting cells which stably express the polypeptide,
• (iv) incubating the cells which stably express the polypeptide in a supernatant, and
• (v) recovering and purifying the polypeptide from the supernatant.