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. 2026 May 6;11(19):28128–28140. doi: 10.1021/acsomega.5c12731

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© 2026 The Authors. Published by American Chemical Society

This article is licensed under CC-BY-NC-ND 4.0

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Cma ST8Sia IV autopolysialylates its own N- and O-glycans. (A) Workflow illustrating the purification steps for Fc-tagged ST8Sia IV. Created with BioRender.com. (B) The recombinant Cma Fc-tagged ST8Sia IV produced in the supernatant of HEK293 and HEK293S cells was isolated with Protein A magnetic beads and visualized by Coomassie staining after SDS-PAGE. (C) Isolated Cma Fc-tagged ST8Sia IV was used for autopolysialylation reactions overnight with or without CMP-Neu5Ac. Reaction products were run on SDS-PAGE and visualized by WB using the anti-polySia mAb735. EndoN-treated samples were used as a negative control. (D) In addition, polysialylated products were treated by PNGase F and/or endoN, run on SDS-PAGE, and visualized by WB with mAb735 against polySia or (E) with an HRP-conjugated goat anti-rabbit IgG antibody against the Fc-tag of Cma ST8Sia IV.