Specificity of human anti-carbohydrate IgG antibodies as probed with polyacrylamide-based glycoconjugates |
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Authors: | EP Smorodin OA Kurtenkov BL Sergeyev GV Pazynina NV Bovin |
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Institution: | (1) Institute of Experimental & Clinical Medicine, Hiiu 42, 11619 Tallinn, Estonia;(2) Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya, 16/10, 117871 Moscow, Russia |
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Abstract: | The TF, Tn, and SiaTn glycotopes are frequently expressed in cancer-associated mucins. Antibodies to these glycotopes were
found in human serum. A set of polyacrylamide (PAA)—based glycoconjugates was applied to the direct and competitive enzyme-linked
immunosorbent assays (ELISA) to characterize the specificity of serum IgG antibodies. The anti-TF, -Tn and -SiaTn IgG were
affinity purified from serum of cancer patients and characterized using PAA-conjugates and free saccharides. The anti-TF and
-Tn antibodies were shown to be specific. The anti-TF IgG bound both Galβ1-3GalNAcα- and Galβ1-3GalNAcβ-PAA, the latter was
three-four times more effective inhibitor of antibody binding. The anti-Tn IgG reacted only with GalNAcα-PAA. The anti-SiaTn
IgG cross-reacted with Tn-PAA but SiaTn-PAA was five-six times more effective inhibitor in a competitive assay. The IC50 values for PAA-conjugates with the corresponding antibodies typically ranged from 2 to 5 × 10−8 M. The antibodies display a low specificity to mucin-type glycoconjugates in comparison with PAA-conjugates as was shown
for mucins isolated from human malignant tumor tissues, ovine submaxillary mucin (OSM) and asialo-OSM. The unusual IgG-antibody
specificity to GalNAcβ and GalNAcβ1-3GalNAcβ ligands was found in human serum. Published in 2004.
This revised version was published online in July 2006 with corrections to the Cover Date. |
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Keywords: | antibodies glycoconjugates mucins TF Tn SiaTn |
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