Functional Validation of LiGA

Functional Validation of LiGA

Comprehensive Functional Validation Ensures Optimal Delivery

CD BioGlyco is a glycobiology company providing various Glycan Display Arrays. Based on the Liquid Glycan Display Array (LiGA), our researchers provide a one-stop service including Construction, Modification, Quality Control, and functional validation. We have developed a wide variety of glycan-modified phage conjugates. Our analytical team has analyzed the structure and density of glycans on phage by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Based on these services, our scientists have developed a LiGA library that includes a wide variety of glycan combinations with different densities and structures. Then, we perform a functional validation for the LiGA.

At CD BioGlyco, we show multiple compounds of different glycan types with the aid of the LiGA library. Depending on the type, we offer the following two services:

  • Functional integrity analysis of glycans on individual glycosylated phages
  • We provide a functional integrity testing service for glycans on individual glycosylated phages via enzyme-linked immunosorbent assay (ELISA). We provide services such as mouse culture, model building, blood collection, and index analysis. Each step is optimized and performed under strict control.
  • Analysis of phage clone mixtures
  • We measure the composition of mixtures of phage clones by plaque-forming units (PFU). Fluorescent plaques of different colors are measured on the bacterial agar layer and provide information on the degree of binding of the associated glycan on the phage. In addition, we provide analytical services for clonal decorative phages fixed with ConA or Galectin-3 (G3C) proteins.
  • We optimize inputs and washes for LiGA binding assays by PFU analysis to detect biotin-tagged proteins or detect whether LiGA mixtures contain target-binding clones. Furthermore, we identify conditions for minimizing nonspecific binding based on PFU assays and provide deep sequencing services for glycan-protein interaction studies.

Fig.1 Schematic representation of functional validation of LiGA. (CD BioGlyco)Fig.1 Schematic representation of functional validation of LiGA. (CD BioGlyco)

Publication

Journal: Nature Chemical Biology

Technology: DNA sequencing, PFU analysis, PCR

Published: 2021

IF: 16.284

Results: In this article, researchers used LiGA (consisting of bacteriophages) to modify the host cells and transduce genes and glycosidases. The α-man modified clones cannot be determined by PFU analysis for binding to ConA alone or LiGA YY (a complex mixture of multiple ligands). This is due to the analytical objective of PFU analysis is not determined. Furthermore, the results showed that Chinese hamster ovary (CHO) cells of mouse CD22 (mCD22) had preferential binding advantages to Neu5Gcα2-6LacNAc due to the known efficient and specific effect of mCD22 on Neu5Gc sialosides. The PFU assay is a critical step in downstream PCR and sequencing. It plays an important role in the LiGA functional validation species.

Fig.2 A colorimetric PFU assay for rapid validation of LiGA. (Sojitra, et al., 2021)Fig.2 A colorimetric PFU assay for rapid validation of LiGA. (Sojitra, et al., 2021)

Applications

  • Functional validation of LiGA can be used to test proteins simultaneously with cells.
  • The LiGA functional validation information plays a crucial role in the study of the molecular interactions between the polysaccharide multivalent display and the multivalent display of cell surface receptors.

CD BioGlyco has a professional analysis team and experienced operators to provide reliable service for our clients. The project report and analysis data will be delivered to clients on time. According to the characteristics of the project, we personalize the adjustment of the service content. Please feel free to contact us.

Reference

  1. Sojitra, M.; et al. Genetically encoded multivalent liquid glycan array displayed on M13 bacteriophage. Nat Chem Biol. 2021, 17(7): 806-816.
This service is for Research Use Only, not intended for any clinical use.

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