A novel radioimmunoassay for human erythropoietin using a synthetic NH2-terminal polypeptide and anti-peptide antibodies.
Antibodies prepared by immunizing rabbits with a synthetic polypeptide representing the first 26 amino acids of the proposed NH2-terminal sequence of human erythropoietin cross-react with peptide, highly purified 125I-labeled erythropoietin and biologically active erythropoietin. We have now developed a rapid and convenient radioimmunoassay for the hormone using 125I-labeled peptide and anti-peptide antibodies. This peptide/anti-peptide approach completely circumvents the relative lack of availability of highly purified 125I-labeled erythropoietin and of erythropoietin for immunization purposes and makes a radioimmunoassay for the hormone universally available to investigators. This concept may be applied as a general method for designing radioimmunoassays of molecules available in severely restricted quantities.
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D000936:Antigen-Antibody Complex / Q000276:immunology; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D004921:Erythropoietin / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D011863:Radioimmunoassay / Q000379:methods*
D000906:Antibodies; D000936:Antigen-Antibody Complex; D010455:Peptides; D004921:Erythropoietin