title: Generation of a human anti-tumor necrosis factor-α monoclonal antibody by in vitro immunization with a multiple antigen peptide.
abstract: We developed the in vitro immunization method to induce antigen-specific immune responses in human peripheral blood mononuclear cells (PBMCs). However, when we used a peptide as sensitizing antigen, the antigen-specific immune response was found to be weak, and hence, we could not effectively obtain the antigen-specific antibody gene. In the present study, we attempted to improve the in vitro immunization method by augmenting the immune response to the peptide antigen. We used a multiple antigen peptide for sensitization. In vitro immunization of the multivalent antigen elicited a strong antigen-specific immune response in the PBMCs, and we succeeded in obtaining antigen-specific antibody genes by the phage-display method. Further, by combining the variable-region genes and constant-region genes of human IgG, we obtained four independent human monoclonal antibodies specific for tumor necrosis factor-α. This might be a good strategy for generating antigen-specific human monoclonal antibodies using a peptide antigen.
authors.affiliation: Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Fujiki
authors.forename: Atsuji
authors.initials: A
authors.lastname: Tsuji
authors.forename: Shin-ei
authors.initials: SE
authors.lastname: Matsumoto
authors.forename: Makiko
authors.initials: M
authors.lastname: Yamashita
authors.forename: Kiichiro
authors.initials: K
authors.lastname: Teruya
authors.forename: Sanetaka
authors.initials: S
authors.lastname: Shirahata
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Katakura
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D005818:Genetic Engineering / Q000379:methods; D006801:Humans; D007114:Immunization / Q000379:methods*; D007127:Immunoglobulin Constant Regions / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000276:immunology; D010455:Peptides / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D007127:Immunoglobulin Constant Regions; D007135:Immunoglobulin Variable Region; D010455:Peptides; D014409:Tumor Necrosis Factor-alpha