Generation and characterization of monoclonal antibodies to human keratinocyte growth factor receptor.
Keratinocyte growth factor receptor (KGFR) and fibroblast growth factor receptor (FGFR) 2c share identical amino acid sequences, except for a 46-amino acid domain in the extracellular region. Monoclonal antibodies (MAbs) specific to KGFR have not been reported nor are commercially available. In this study, we generated murine MAbs specific to KGFR in non-obese diabetic (NOD) mice using a modified Repeated Immunizations at Multiple Sites (RIMMS) technology. Stable cell lines expressing the full-length human KGFR or FGFR2c were produced to facilitate the identification of KGFR-specific MAbs. Following the initial screening of hybridoma clones with a fluorescence-based, confocal cell detection method and ELISA, KGFR-specific MAbs were selected and confirmed by flow cytometry and Western blot analyses. Antagonistic MAbs were identified using a cell-based functional assay. These KGFR MAbs will be important reagents for studying the biological function and tissue distribution of this receptor in normal and pathological conditions.
Wei
Ping
P
Department of Hematology, Amgen Inc., Thousand Oaks, California 91320-1789, USA. pwei@amgen.com
Zhan
Jinghui
J
Liu
Shuying
S
Chang
David
D
Haldankar
Raj
R
Burkhardt
Kim
K
Crouse
Jill
J
Hui
John
J
Juan
Todd
T
Talvenheimo
Jane
J
Kim
Helen
H
Li
Luke
L
Boone
Tom
T
Borges
Luis
L
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry*; D000918:Antibody Specificity; D015153:Blotting, Western; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D005434:Flow Cytometry; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D051497:Receptor, Fibroblast Growth Factor, Type 2 / Q000276:immunology*
D000911:Antibodies, Monoclonal; D051497:Receptor, Fibroblast Growth Factor, Type 2; C065104:keratinocyte growth factor receptor