Expression in yeast of amino-terminal peptide fusions to hepatitis B core antigen and their immunological properties.
Hepatitis B core protein (HBcAg) is a potent antigen that gives both a T-cell-dependent and a T-cell-independent antibody response. It has been shown that a foreign epitope can be fused to the amino terminus of HBcAg without affecting particle integrity, and that the resulting chimaeric cores retain the immunogenicity of the foreign epitope. Here we describe the efficient expression in yeast of two different chimaeric cores, carrying epitopes of Foot and Mouth Disease Virus (FMDV) or human chorionic gonadotrophin (hCG), which are candidates for FMD and contraceptive vaccines, respectively. These cores could not be produced in E. coli in soluble form but were expressed to high levels in yeast. We constructed a yeast expression vector that allows rapid production of different chimaeric cores by cloning in cassettes encoding foreign epitopes. Both FMDV and hCG-cores were shown to present the epitopes at the surface of the particles. The FMDV-cores produced in yeast were efficient inducers of neutralising antibodies in guinea-pigs after one low dose.
Beesley
K M
KM
Department of Molecular Biology and Virology, Wellcome Research Laboratories, Beckenham, Kent.
Francis
M J
MJ
Clarke
B E
BE
Beesley
J E
JE
Dopping-Hepenstal
P J
PJ
Clare
J J
JJ
Brown
F
F
Romanos
M A
MA
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D005537:Aphthovirus / Q000276:immunology; D001483:Base Sequence; D002678:Chimera; D006063:Chorionic Gonadotropin / Q000235:genetics; D004271:DNA, Fungal; D000939:Epitopes / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D005822:Genetic Vectors; D006168:Guinea Pigs; D006512:Hepatitis B Core Antigens / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D006063:Chorionic Gonadotropin; D004271:DNA, Fungal; D000939:Epitopes; D006512:Hepatitis B Core Antigens; D011993:Recombinant Fusion Proteins