Oligosaccharides as Immunodeterminants of Erythropoietin for Two Sets of Anti-Carbohydrate Antibodies

Abstract

Antibodies directed against recombinant erythropoietin have been obtained by immunization of rabbits with the hormone in Freund's complete adjuvant. Two sets of antibodies are present in the serum of the immunized rabbits. The results of oxidation of the erythropoietin with periodate, inhibition of the antibodies with the structural monosaccharide residues of the hormone, and reaction of the antibodies with lectins of known carbohydrate specificity have established the antibodies to be anti-carbohydrate antibodies. These antibodies may be of value as tracking agents for some diseases and should be useful for detecting abuses of the hormone in enhancing performance in athletic competitions.

Efficient purification and preconcentration of erythropoietin in human urine by reusable immunoaffinity column

“…N-Hydroxysuccinimide (NHS), N-ethyl-N -(3-dimethylaminopropy)carbodimide (EDC), N-2-hydroxyethylpiperazine-N -2-ethanesulfonic acid (HEPES) and carboxymethyl dextran (CM5) sensorchip were purchased from Biacore Inc. (Uppsala, Sweden). rHuEPO was modified by oxidizing the carbohydrate portion with sodium periodate and keeping the polypeptide chain intact for specificity test [19].…”

“…Since EPO is a kind of glycoprotein and consists of peptide portion and carbohydrates, NaIO 4 can be used to destroy the structure of carbohydrates via oxidation and keep the peptide portion intact [19]. To determine the specific part of EPO that the antibodies selectively bound to, cross-reaction of NaIO 4 -treated rHuEPO was investigated and the results were shown in Fig.…”