Preparation of asialo-agalacto-glycophorin A for screening of anti-Tn antibodies.

Abstract

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.

• Authors: Tomoyuki Koizumi, Ayano Matsumoto-Takasaki, Hiroshi Nakada, Munehiro Nakata, Yoko Fujita-Yamaguchi
• Journal: Bioscience trends (Biosci Trends) Vol. 4 Issue 6 Pg. 308-11 (Dec 2010) ISSN: 1881-7823 [Electronic] Japan
• PMID: 21248428 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies
• Antibodies, Monoclonal
• Antigens, Tumor-Associated, Carbohydrate
• Epitopes
• Glycophorins
• Oligosaccharides
• Peptide Library
• Polysaccharides
• Tn antigen
• Neuraminidase
• beta-Galactosidase
• N-Acetylneuraminic Acid

Topics

• Antibodies (chemistry)
• Antibodies, Monoclonal (chemistry)
• Antigens, Tumor-Associated, Carbohydrate (chemistry, immunology)
• Enzyme-Linked Immunosorbent Assay (methods)
• Epitopes (chemistry)
• Glycophorins (chemistry)
• Humans
• N-Acetylneuraminic Acid (chemistry)
• Neoplasm Metastasis
• Neoplasms (immunology, therapy)
• Neuraminidase (chemistry)
• Oligosaccharides (chemistry)
• Peptide Library
• Polysaccharides (chemistry)
• beta-Galactosidase (metabolism)