Formation of immunogenic virus-like particles by inserting epitopes into surface-exposed regions of hamster polyomavirus major capsid protein.
We generated highly immunogenic virus-like particles that are based on the capsid protein VP1 of the hamster polyomavirus (HaPV-VP1) and harbor inserted foreign epitopes. The HaPV-VP1 regions spanning amino acids 81-88 (position 1), 222/223 (2), 244-246 (3), and 289-294 (4) were predicted to be surface exposed. An epitope of the pre-S1 region of the hepatitis B virus (designated S1; amino acid sequence DPAFR) was introduced into the predicted positions of VP1. All VP1/S1 fusion proteins were expressed in yeast and generated virus-like particles. Immunoassays using the S1-specific monoclonal antibody MA18/7 and immunization of C57Bl6 mice with different VP1/S1 constructs showed a pronounced reactivity and a strong S1-specific antibody response for particles carrying the insert in position 1, 2, 1+2, and 1+3. Our results suggest that HaPV-VP1 represents a highly flexible carrier moiety for the insertion of foreign sequences offering a broad range of potential uses, especially in vaccine development.
Gedvilaite
A
A
Institute of Biotechnology, Vilnius, LT-2028, Lithuania.
Frömmel
C
C
Sasnauskas
K
K
Micheel
B
B
Ozel
M
M
Behrsing
O
O
Staniulis
J
J
Jandrig
B
B
Scherneck
S
S
Ulrich
R
R
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D002213:Capsid / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D036022:Capsid Proteins; D006224:Cricetinae; D017248:Enzyme Multiplied Immunoassay Technique; D000939:Epitopes / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D016253:Microscopy, Immunoelectron; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000235:genetics*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011120:Polyomavirus / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D011487:Protein Conformation; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016415:Sequence Alignment; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D000939:Epitopes; D006514:Hepatitis B Surface Antigens; D010446:Peptide Fragments; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C062481:VP1 protein, polyomavirus; D014614:Vaccines, Synthetic; C050806:presurface protein 1, hepatitis B surface antigen