Identification of the insulin-like growth factor I (IGF I) epitopes recognized by monoclonal and polyclonal antibodies to IGF I.
We have characterized the binding epitopes of human insulin-like growth factor I (IGF I) for a polyclonal (UB286) and a monoclonal (SM 1.2) antibody using IGF analogs obtained by site-directed mutagenesis. The polyclonal antibody, UB286, which was obtained from the National Hormone and Pituitary Program, recognizes determinants surrounding residues 15 and 16 in the B-region and residues 49-51, 55 and 56 in the A-region. These residues are predicted to be within helical segments which are accessible for surface binding. The monoclonal antibody SM 1.2 selectively recognizes the region surrounding residues 15 and 16. Antibodies UB286 and SM 1.2 are both neutralizing antibodies as judged by their ability to inhibit binding of 125I-IGF I to type 1 receptors on human placental membranes. In addition, SM 1.2 inhibits the ability of IGF I and IGF analogs for which it has high affinity to stimulate DNA synthesis in murine fibroblasts. In contrast, analogs with substitutions at residues 15 and 16, which have poor affinity for SM 1.2, stimulate DNA synthesis with equal potency in the presence and absence of SM 1.2. These antibodies bind normally to analogs which we have previously shown have drastically reduced binding to type 1 IGF receptors, indicating that the antibodies and the receptors recognize distinct domains of IGF I.
Department of Biochemical Endocrinology, Merck Sharp and Dohme Research Laboratories, Rahway, New Jersey 07065.
M A
MA
Cascieri
M L
ML
Bayne
E
E
Ber
B G
BG
Green
G W
GW
Men
G G
GG
Chicchi
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology* / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000494:pharmacology; D000915:Antibody Affinity; D000918:Antibody Specificity; D001667:Binding, Competitive; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000031:analogs & derivatives / Q000276:immunology* / Q000378:metabolism; D008969:Molecular Sequence Data; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D011956:Receptors, Cell Surface / Q000378:metabolism; D017451:Receptors, Somatomedin; D013002:Somatomedins / Q000276:immunology*; D013329:Structure-Activity Relationship
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D011956:Receptors, Cell Surface; D017451:Receptors, Somatomedin; D013002:Somatomedins; D007334:Insulin-Like Growth Factor I