Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation

doi:10.1016/0022-1759(85)90337-0

Journal of Immunological Methods

Volume 78, Issue 1, 8 April 1985, Pages 143-153

https://doi.org/10.1016/0022-1759(85)90337-0 Get rights and content

Abstract

A method is described for determining whether particular monoclonal antibodies are specific for carbohydrate or non-carbohydrate antigenic determinants. In a model system consisting of the Lewis a human blood group determinant attached to either protein or lipid, mild periodate oxidation destroyed the carbohydrate determinant without altering protein or lipid epitopes. The technique was readily applied to antigens bound to plastic wells for ELISA, to nitrocellulose sheets for Western blots, and to thin layer chromatography (TLC) plates for TLC immunostaining. Mild periodate oxidation can prove useful during the early stages of hybridoma screening in order to select for or against anti-carbohydrate antibodies.

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This dynamic nature favors self-healing [21] and injectability, thus imine-crosslinked hydrogels have shown potential in applications like sequential drug delivery [7] and tissue engineering. Bloodgood [22] reported the preparation of imine-based hydrogels from polysaccharides utilizing sodium periodate to oxidize the vicinal hydroxyl groups on C2 and C3, thereby creating ring-opened dialdehydes (Figure 3a). Any aldehyde groups within the hydrogel not involved in crosslinking imine bonds can be used for conjugating active small molecules that possess appropriate chemical moieties.
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Fig. 2 presents ELISA data obtained with two synthetic glycopeptides representing two different sections of the extracellular domain of the aGP molecule, arbitrarily called glycopeptides A and B (Table 2B). Fig. 2A demonstrates that binding is carbohydrate-dependent, since both the glycopeptide A after treatment with periodate under carbohydrate-specific conditions according to [15], which destroys the terminal Gal, as well as the nonglycosylated peptide are completely negative in ELISA. Fig. 2B shows that both glycopeptides are recognized by the anti-TF antibodies, demonstrating that their epitopes do not involve parts of the peptide sequence.
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Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation

Abstract

Clin. Chim. Acta

Adv. Carbohydrate Chem. Biochem.

J. Biol. Chem.

Science

Biochem. J.

J. Exp. Med.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

J. Natl. Cancer Inst.

J. Immunol.

Biochem. J.

Structural Concepts in Immunology and Immunochemistry

Nature (London)