Discovery of indole-modified aptamers for highly specific recognition of protein glycoforms

Abstract Glycosylation is one of the most abundant forms of post-translational modification, and can have a profound impact on a wide range of biological processes and diseases. Unfortunately, efforts to characterize such modifications in the context of basic and clinical research are severely hampe...

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Bibliographic Details
Published inbioRxiv
Main Authors Yoshikawa, Alex M, Rangel, Alexandra, Feagin, Trevor, Chun, Elizabeth, Leighton, Wan, Li, Anping, Moekl, Leonhard, Eisenstein, Michael, Pitteri, Sharon, Soh, H Tom
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 14.03.2021
Subjects
Affinity
Aptamers
Epitopes
Flow cytometry
Glycosylation
Indoles
Post-translation
Proteins
Reagents
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Summary:Abstract Glycosylation is one of the most abundant forms of post-translational modification, and can have a profound impact on a wide range of biological processes and diseases. Unfortunately, efforts to characterize such modifications in the context of basic and clinical research are severely hampered by the lack of affinity reagents that can differentiate protein glycoforms. This lack of reagents is largely due to the challenges associated with generating affinity reagents that can bind to particular glycan epitopes with robust affinity and specificity. In this work, we use a fluorescence-activated cell sorting (FACS)-based approach to generate and screen aptamers with indole-modified bases in an effort to isolate reagents that can differentiate between protein glycoforms. Using this approach, we were able to select multiple aptamers that exhibit strong selectivity for specific glycoforms of two different proteins, with the capacity to discriminate between molecules with identical tertiary structures that differ only in terms of their glycan modifications. Competing Interest Statement The authors have declared no competing interest.
DOI:10.1101/2021.03.13.435263