Preparation of asialo-agalacto-glycophorin A for screening of anti-Tn antibodies.
Oncogenic antigens such as Tn-antigen (GalNAcα-Ser/Thr) are involved in metastatic processes and are associated with a poor prognosis, thus representing excellent targets for cancer intervention. Available anti-Tn antibodies which can be applied for therapeutics or diagnostics are severely limited mostly because the Tn-antigen epitope by itself is too small to be antigenic in addition to the fact that many carbohydrates are self-antigens. To characterize anti-Tn monoclonal antibodies as well as to perform panning and screening for isolation of anti-Tn single chain variable fragments from phage-display libraries, a large quantity of inexpensive Tn-antigens are needed. In this study, thus, glycophorin A which is a highly glycosylated sialoglycoprotein with approximately 12 O-glycans was sequentially treated with sialidase and β-galactosidase to remove sialic acid and galactose residues. The resulted product was shown to be an asialo-agalacto-glycophorin A which is reactive to an anti-Tn-antigen antibody. The simple preparation procedures described here would greatly help production and characterization of potentially valuable anti-Tn-antigen antibodies, which can be readily developed for cancer therapeutics and diagnostics.
Department of Applied Biochemistry, Tokai University, Kanagawa, Japan.
Tomoyuki
T
Koizumi
Ayano
A
Matsumoto-Takasaki
Hiroshi
H
Nakada
Munehiro
M
Nakata
Yoko
Y
Fujita-Yamaguchi
D000906:Antibodies / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000737:chemistry* / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000737:chemistry; D006021:Glycophorins / Q000737:chemistry; D006801:Humans; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D009362:Neoplasm Metastasis; D009369:Neoplasms / Q000276:immunology / Q000628:therapy; D009439:Neuraminidase / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry; D019151:Peptide Library; D011134:Polysaccharides / Q000737:chemistry; D001616:beta-Galactosidase / Q000378:metabolism
D000906:Antibodies; D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; D000939:Epitopes; D006021:Glycophorins; D009844:Oligosaccharides; D019151:Peptide Library; D011134:Polysaccharides; C040286:Tn antigen; D009439:Neuraminidase; D001616:beta-Galactosidase; D019158:N-Acetylneuraminic Acid