Fig. 5: A mass spectrometry assay to detect and quantify Siglec-glycan interactions.
From: A versatile soluble siglec scaffold for sensitive and quantitative detection of glycan ligands
a SDS-PAGE of Siglec-1-Fc following digestion with TEV and Endo-H. b Mass spectrum for Siglec-1 produced from WT and Lec1 CHO cells as the full Fc-chimera, following digestion with TEV, and after treatment with Endo-H. c, d Mass spectrum of CD22 (c) and CD33 (d) from Lec1 CHO cells treated with TEV and Endo-H. e ESI mass spectra of 3.6 μM Siglec-1 in its unliganded form (upper) and complexed with 80 μM Neu5Acα2-3Lac (2) (lower). Spectra are representative of at least two independent replicates. Dashed lines are a guide to line up the same speak in each spectrum. f Summary of affinities (KD) of the four trisaccharides for the fragments of Siglec-1, CD22, and CD33.
