Production and characterization of monoclonal antibodies to human sclerostin.
We developed and characterized monoclonal antibodies directed against the amino-terminal and carboxy-terminal regions of human and mouse sclerostin (scl). Amino-terminal and carboxy-terminal scl peptides with limited homology to scl domain-containing protein-1 were synthesized using f-moc chemistry. The peptides were conjugated to keyhole limpet hemocyanin and the conjugates were used for immunization of mice. Monoclonal antibodies were obtained and characterized using bacterially expressed and insect cell-expressed recombinant scl. The amino-terminal (IgG 2aK) and carboxy-terminal (IgG 2bK) antibodies bound bioactive sclerostin that was expressed in an insect-cell expression system with dissociation constants in the nanomolar range. The antibodies are potentially useful agents that can be used for modulating sclerostin bioactivity.
Craig
Theodore A
TA
Nephrology Research, Department of Medicine, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, Minnesota 55905, USA.
Sommer
Stacy L
SL
Beito
Thomas G
TG
Kumar
Rajiv
R
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D001483:Base Sequence; D019485:Bone Morphogenetic Proteins / Q000276:immunology*; D002478:Cells, Cultured; D017931:DNA Primers; D005819:Genetic Markers / Q000276:immunology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D020349:Surface Plasmon Resonance
D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; D019485:Bone Morphogenetic Proteins; D017931:DNA Primers; D005819:Genetic Markers; C422903:SOST protein, human