Generation and characterization of new monoclonal antibodies against human chymase.
We have succeeded in producing monoclonal antibodies directed against a wide variety of epitopes of human chymase by using two different immunogens: a recombinant human chymase-heparin mixture, and chymase alone. Hybridomas were screened by ELISA, and 7 clones were selected based on antibody titers. Epitopes were localized by Western blotting with a C-terminal-deletion series of chymase-GST fusion proteins, and it was possible to use the antibodies for Western blotting and immunohistochemistry. Dot-blot analysis for species specificity revealed that the MAbs bound canine chymase as well as human chymase, and that two of them also bound rodent chymases. These results indicate that the antibodies can be used for various immunological analyses in further investigations of chymase.
Kunori
Yuichi
Y
Pharmaceutical Discovery Research Laboratories, TEIJIN Institute for Biomedical Research, Hino, Tokyo, Japan. y.kunori@teijin.co.jp
Hase
Naoki
N
Kawamura
Takashi
T
Sato
Hiroaki
H
Kasai
Hidenori
H
Muroga
Yumiko
Y
Kamimura
Takashi
T
Fukamizu
Akiyoshi
A
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western; D053818:Chymases; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006801:Humans; D007150:Immunohistochemistry; D012697:Serine Endopeptidases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D012697:Serine Endopeptidases; D053818:Chymases