LiGA Construction Service

LiGA Construction Service

Attentive and Rigorous Service Related to LiGA Construction

CD BioGlyco provides LiGA analysis services for client studies on glycan-protein interactions, etc. LiGA combines the multivalent presentation of liposome constructs, conventional arrays, and DNA-encoded glycan density and composition. LiGA construction is a key step in the entire process of LiGA analysis. Based on our well-established Glycan Display Platform, we provide high-quality LiGA construction services. To make our services more efficient, we optimize our cloning methods and use them in conjunction with a variety of chemical and enzymatic coupling methods.

We use M13 bacteriophage as a stable and scalable platform for constructing LiGA. M13 virus particles are not naturally glycosylated. We couple the glycans required for the LiGA array to a subset of copies of the major capsid protein pVIII, which produces a multivalent display of glycans. The coupling ultimately produces a bacteriophage population with a single glycan composition. With this strategy, we assemble bacteriophage glycan libraries with different DNA coding barcodes. Each barcode has a different glycan and is used for studies such as protein-glycan interactions. In this process, we remove unsuitable clones to ensure the accuracy of the experiment.

Our experimental steps included the following process.

We clone the degenerate nucleotide sequence into the bacteriophage genome near the pIII cloning site. This results in a library of silenced barcode vectors. These vectors encode chemically identical bacteriophage particles.

Before doping bacteriophage, we synthesize multiple glycans or screen natural sources of glycans by chemoenzymatic or chemical methods. After that, we use chemical and enzymatic methods to link glycans to bacteriophage. Then, we obtain glycan modified bacteriophage library.

  • Characterization of LiGA at CD BioGlyco

We are equipped with advanced equipment such as matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to analyze the density of glycans on the bacteriophage surface. Meanwhile, we also verify the functionality of LiGA to ensure the reliability and functional integrity of LiGA.

Fig.1 Process of LiGA construction. (CD BioGlyco)Fig.1 Process of LiGA construction. (CD BioGlyco)

Publication Data

Technology: Gene editing, Chemical conjugation

Journal: Nature Chemical Biology

IF: 16.174

Published: 2021

Results: In this study, synthetic glycans with alkyl-azido linkers were attached to the M13 bacteriophage. Finally, the polymer of M13 bacteriophage containing about 140 glycan modifications was successfully constructed. Validation of the functionality of the constructed LiGA showed that the LiGA could be used to detect glycan binding profiles on cells in vivo and in vitro. In addition to this, glycan affixes with optimal affinity necessary for lectin binding were identified. The LiGA remedies the shortcomings of slide-based glycan-based arrays.

Fig.2 Synthesis and characterization of LiGA components. (Sojitra, et al., 2021)Fig.2 Synthesis and characterization of LiGA components. (Sojitra, et al., 2021)

Applications of LiGA Construction

  • The construction of LiGA plays an important role in analyzing the glycan binding properties of proteins, including animal and plant lectins, growth factors, and antibodies.
  • The construction of LiGA plays an important role in probing cell-glycan interactions in vivo and in vitro.
  • The construction of LiGA is used in studies to improve the design of inhibitors, vaccines, and therapeutic drugs.

Highlights of Us

  • We optimize every aspect of LiGA construction, including the construction of M13 bacteriophage libraries, glycan coupling, and more.
  • Clients just need to tell us the requirements and we complete all the experiments related to LiGA construction.
  • The density of polysaccharide multivalent display affect the specificity and affinity of the binding of carbohydrate processing enzymes, lectins, antibodies, and so on. We promptly adjust the density of multiple displays to ensure the accuracy of the results.

CD BioGlyco has a well-established LiGA platform for the detection of many forms of samples. Please feel free to contact us to learn more about LiGA, LiGA construction, and other related experimental content. With our extensive experience in LiGA construction, we will perform various assays with excellence.

Reference

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

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