A novel antibody AA98 V(H)/L directed against CD146 efficiently inhibits angiogenesis.
BACKGROUND
An anti-CD146 monoclonal antibody, AA98, has been identified as an inhibitor of tumor angiogenesis. To overcome the inherent immunogenicity of murine antibody as well as to facilitate immunotoxin construction, a single chain AA98 V(H)/L with three-domain fragments was constructed and expressed in mammalian cells.


METHODS
The genes of the AA98 heavy chain variable region and the light chain were linked with a modified 12 amino acid sequence that was derived from the heavy chain C(H)1 region, thus constituting the three-domain antibody V(H)/L. Soluble AA98 V(H)/L was produced by mammalian cells and purified by affinity chromatography. The specificity of AA98 V(H)/L for the CD146 molecule was detected by ELISA, immunofluorescence staining and flow cytometry.


RESULTS
AA98 V(H)/L alone showed anti-angiogenic properties in a chicken chorioallantoic membrane (CAM) assay as the parent mAb AA98 did.


CONCLUSIONS
This newly generated AA98 V(H)/L antibody displays a therapeutic potential for tumor and other angiogenesis disorders, as well as providing a new strategy for antibody engineering for clinical applications.
Lin
Yun
Y
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, PR China.
Wu
Xiaoping
X
Shen
Yi
Y
Bu
Pengcheng
P
Yang
Dongling
D
Yan
Xiyun
X
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D051929:CD146 Antigen / Q000276:immunology; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply; D042783:Endothelial Cells / Q000166:cytology / Q000276:immunology*; D000939:Epitopes; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology*; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D018919:Neovascularization, Physiologic / Q000276:immunology
D000911:Antibodies, Monoclonal; D051929:CD146 Antigen; D000939:Epitopes; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; C497902:MCAM protein, human