CD BioGlyco has extensive experience in Liquid Glycan Display Array (LiGA). The multivalent glycan display is generated on phage by glycanization modification. These detailed services for probing cell surface receptors using LiGA include preparation, modification, assembly of LiGA library, and high-throughput screening of glycan surface receptors. Siglec belongs to the Ig superfamily (type I lectins), which is mainly expressed on leukocytes except for tumor cells and participates in immune responses related to tumors and microbial infections by recognizing glycans containing sialic acid. Our researchers analyze the linkage specificity of lectin ligands by using cell lines with different glycosylation levels on surfaces.
Fig.1 Flow chart of a typical binding assay between cells and LiGA library. (CD BioGlyco)
We have optimized the key parameters of the LiGA-based cell surface lectin-binding profiling service. We perform polymerase chain reaction (PCR) amplification using primers and isogenic cell lines. Moreover, we provide deep-sequencing and differential enrichment analysis services.
Our researchers have analyzed in detail the biodistribution of LiGA in organs in mice and its association with two subpopulations of B cells. We offer detection services of interactions between lectin ligands and receptors displayed on organ and cell surfaces in living animals. After enrichment, B-cell populations are isolated from the organs. Plaque forming units (PFU) assay service helps clients to analyze the composition of libraries from different organs with B-cell types.
Paper Title: Measuring carbohydrate recognition profile of lectins on live cells using liquid glycan array (LiGA)
Technology: Next-generation sequencing (NGS), Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), PCR
Published: 2023
Results: In this study, five members of the Siglec family were evaluated using LiGA-100 and a focused glycan library LiGA-g. The glycans were tested for specificity by their expression on different cell lines. Their expression on different cell lines tested glycan binding specificity. A fraction of the 19 glycans enriched by Siglec-9+ could be enriched by Siglec-7+ cells or CD22+ Chinese hamster ovary (CHO) cells. In addition, Siglec-9 was found to bind more to sialic acid for α2-3 linked sialic acid than α2-6 linked. By analyzing LiGA-100 binding, it was observed that α2+ cells were slightly specific for α2-3 linked sialic acid glycans connected to Galβ1-3Glc (GM3).
Fig.2 Data on the binding intensity of the three ligands on different cell surfaces. (Sojitra, et al., 2023)
CD BioGlyco has established a high-throughput screening Glycan Display Platform. Our talented team will help clients plan the entire program of services. To learn more information about LiGA-based cell surface lectin-binding profiling, please feel free to contact us.
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