Monoclonal antibodies to the amino- and carboxyl-terminal domains of ovotransferrin.

Monoclonal antibodies to the iron transport protein ovotransferrin were produced by immunizing mice simultaneously with ovotransferrin and with the proteolytically derived amino- and carboxyl-terminal half-molecule domains of ovotransferrin. Two isolated hybridoma clones (designated alpha OT + N1 and alpha OT + N2) produced antibodies (IgG1) to determinants located on holo-ovotransferrin and the amino-terminal domain; two hybridoma clones (designated alpha OT + C1 and alpha OT + C2) produced antibodies (IgG1) to determinants on holo-ovotransferrin and the carboxyl-terminal domain. One hybridoma clone (designated alpha OT-N1) produced an antibody (IgG1) that bound only the amino-terminal domain and did not bind holo-ovotransferrin. Both alpha OT + N1, and alpha OT-N1 bound to antigen less tightly after removal of iron; antibodies alpha OT + N2, alpha OT + Cl, and alpha OT + C2 were unaffected by removal of iron from holo-ovotransferrin or the isolated domains. Intact disulfide bonds in the antigens were required for binding by the antibodies. These antibodies should prove useful as probes for discrete regions of the ovotransferrin molecule, in particular, those regions involved in binding to the transferrin receptor.