Mouse monoclonal antibodies to hepatitis B virus preS1 produced after immunization with recombinant preS1 peptide.
We have efficiently generated mouse monoclonal antibodies (MAbs), which bind specifically to amino acids 21-47 of the preS1 domain of hepatitis B virus (HBV) by immunizing mice with the preS1 peptide (amino acids, aa 1-56) conjugated to keyhole limpet hemocyanin. Hybridomas were screened by an indirect enzyme-linked immunosorbent assay (ELISA) using the purified preS1 peptide as a coated antigen. Eighteen positive hybridomas were selected and subjected to isotyping. Of these, 5 clones secreted immunoglobulin G (IgG) and 13 clones secreted IgM. Four (KR1, KR2, KR3, and KR4) of the 5 IgG MAbs bound to preS1 peptide (aa 21-47). Epitope mapping using bacterially expressed GST fusion proteins revealed that three clones (KR2, KR3, KR4) (IgG1, K) recognize aa 21-35, while KR1 (IgG2a, K) recognizes aa 35-47 of the preS1. These MAbs immunoprecipitated HBV particles, demonstrating that they bind to native HBV particles.
Ryu
C J
CJ
Antibody Engineering Research Unit, Korea Research Institute of Bioscience and Biotechnology, Yuseong, Taejon.
Kim
Y K
YK
Hur
H
H
Kim
H S
HS
Oh
J M
JM
Kang
Y J
YJ
Hong
H J
HJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D018604:Epitope Mapping; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000276:immunology*; D006825:Hybridomas / Q000737:chemistry; D016056:Immunodominant Epitopes / Q000737:chemistry; D007132:Immunoglobulin Isotypes / Q000032:analysis; D051379:Mice / Q000276:immunology; D010455:Peptides / Q000276:immunology; D011233:Precipitin Tests; D011498:Protein Precursors / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014771:Virion / Q000276:immunology
D000911:Antibodies, Monoclonal; D006514:Hepatitis B Surface Antigens; D016056:Immunodominant Epitopes; D007132:Immunoglobulin Isotypes; D010455:Peptides; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C050806:presurface protein 1, hepatitis B surface antigen; D005982:Glutathione Transferase