Screening of carbohydrate-specific phage antibodies against recombinant human erythropoietin (rhuEPO) using a phage display antibody library: preliminary study.
This paper is a preliminary report on development of a screening method for carbohydrate-specific phage antibodies against recombinant human erythropoietin (rHuEPO), using a phage display antibody library. rHuEPO was oxidized with sodium periodate or treated with 1,4-dithiothreitol and guanidine hydrochloride for detecting the specificity of obtained phage antibodies. Of 100 phage clones, three initially showed higher carbohydrate-related specificity. One of them (No. 62) bound specifically to the carbohydrate chains of rHuEPO, while the other two (Nos. 63 and 83) might recognize the steric conformation related to both the carbohydrate and the polypeptide chain of rHuEPO. These phage antibodies may serve as useful capture ligands for future development of efficient analytical methods for rHuEPO.
Mi
Jiebo
J
Institute of Analytical Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing 100 871, PR China.
Ding
Xiaojie
X
Zhao
Meiping
M
Chang
Wenbao
W
Guo
Zhenquan
Z
D000906:Antibodies / Q000235:genetics; D000918:Antibody Specificity; D002241:Carbohydrates / Q000737:chemistry / Q000276:immunology*; D004229:Dithiothreitol; D004797:Enzyme-Linked Immunosorbent Assay; D004921:Erythropoietin / Q000737:chemistry / Q000276:immunology*; D006031:Glycosylation; D019791:Guanidine; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics; D066298:In Vitro Techniques; D010084:Oxidation-Reduction; D019151:Peptide Library; D010504:Periodic Acid; D011994:Recombinant Proteins
D000906:Antibodies; D002241:Carbohydrates; D007128:Immunoglobulin Fragments; D019151:Peptide Library; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D010504:Periodic Acid; D004921:Erythropoietin; C009288:metaperiodate; D019791:Guanidine; D004229:Dithiothreitol