Development of human-human hybridoma from anti-Her-2 peptide-producing B cells in immunized NOG mouse.
OBJECTIVE
Numerous monoclonal antibodies have been developed for the purpose of medical treatments, including cancer treatment. For clinical application, the most useful are human-derived antibodies. In this study, we tried to prepare designed antigen-specific antibodies of completely human origin using immunodeficient mouse.


METHODS
Nonobese diabetic/severe combined immunodeficient/IL-2 receptor gamma null mouse (NOG) mouse was used to reconstitute the human immune system with umbilical cord blood hematopoietic stem cells (CB-NOG mouse) and to prepare human-derived Her-2-epitope-specific antibodies. Hybridoma lines were prepared by fusing the human myeloma cell line Karpas707H.


RESULTS
Serum of immunized NOG mouse contained human-derived immunoglobulin M (IgM) antibodies specific for a short peptide sequence of 20 amino acids, including the epitope peptide of apoptotic Her-2 antibody CH401. Hybridoma lines were successfully prepared with spleen B cells obtained from the immunized CB-NOG mouse. One of these cell lines produced human IgM against the epitope peptide that can recognize surface Her-2 molecule.


CONCLUSIONS
We could produce human-derived IgM antibody against Her-2 epitope peptide in CB-NOG mouse, succeeding in generation of human hybridoma-secreting IgM against a given peptide.
Kametani
Yoshie
Y
Department of Immunology, Tokai University School of Medicine, Kanagawa, Japan.
Shiina
Masashi
M
Katano
Ikumi
I
Ito
Ryoji
R
Ando
Kiyoshi
K
Toyama
Kanae
K
Tsukamoto
Hideo
H
Matsumura
Takuya
T
Saito
Yuki
Y
Ishikawa
Dai
D
Taki
Takao
T
Ito
Mamoru
M
Imai
Kohzoh
K
Tokuda
Yutaka
Y
Kato
Shunichi
S
Tamaoki
Norikazu
N
Habu
Sonoko
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000627:therapeutic use; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology*; D036101:Cord Blood Stem Cell Transplantation / Q000379:methods; D018985:Epitopes, B-Lymphocyte / Q000276:immunology* / Q000494:pharmacology; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology*; D007114:Immunization; D007075:Immunoglobulin M / Q000276:immunology* / Q000627:therapeutic use; D051379:Mice; D008817:Mice, Mutant Strains; D009369:Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000276:immunology; D010455:Peptides / Q000276:immunology* / Q000494:pharmacology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*; D018183:Transplantation Chimera / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018985:Epitopes, B-Lymphocyte; D007075:Immunoglobulin M; D010455:Peptides; D018719:Erb-b2 Receptor Tyrosine Kinases