3 years ago

A new strategy for metal labeling of glycan structures in antibodies

Michael W. Linscheid, Lena Ruhe, Stefanie Ickert, Sebastian Beck

Abstract

Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. In this work, we have developed a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies. The oligosaccharide glycans were selectively trimmed to the last N-acetylglucosamine to which an artificial azide containing galactose residue was bound. This azide can be used for subsequent cycloaddition of an alkyne. Therefore, we developed a lanthanide-containing macrocyclic reagent to selectively connect to this azido galactose. In summary, the glycan structures of an antibody can be labeled with a metal functionality using this approach. Furthermore, the functionality of the antibodies can be fully maintained by labeling the Fc glycans instead of using labeling reagents that target amino or thiol groups. This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples.

Graphical abstract

Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry.

Publisher URL: https://link.springer.com/article/10.1007/s00216-017-0683-1

DOI: 10.1007/s00216-017-0683-1

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