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Glycobiophysics pp 259–273Cite as

Quantifying Weak Glycan-Protein Interactions Using a Biolayer Interferometry Competition Assay: Applications to ECL Lectin and X-31 Influenza Hemagglutinin

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Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1104))

Abstract

This chapter introduces two formats using bio-layer interferometry competition assays to determine the solution K D values of weak glycan-protein interactions. This approach overcomes the challenge of determining weak interactions while minimizing the amount of reagents required. Accurate solution K D values aid in understanding the complex relationships between monomeric versus multimeric interactions and affinity versus avidity. The assays have been applied to a well-studied lectin (Erythrina crista-galli lectin) and influenza hemagglutinin (X-31). The solution K D values determined from this approach are in good agreement with previous reported literature values from isothermal titration calorimetry and NMR. Additionally, this approach appears robust and precise.

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Abbreviations

BLI:

Bio-layer interferometry

CFG:

Consortium for Functional Glycomics

ECL:

Erythrina crista-galli lectin

ELISA:

Enzyme-linked immunosorbent assay

HA:

Hemagglutinin

ITC:

Isothermal titration calorimetry

MST:

Microscale thermophoresis

SPR:

Surface plasmon resonance

NA:

Neuraminidase

NMR:

Nuclear magnetic resonance spectroscopy

3D:

Three-dimensional

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Author information

Authors and Affiliations

  1. Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA

    Ye Ji & Robert J. Woods

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  1. Ye Ji
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  2. Robert J. Woods
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Correspondence to Robert J. Woods .

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Editors and Affiliations

  1. Synthetic Cellular Chemistry Laboratory, RIKEN Cluster for Pioneering Research, Wako, Saitama, Japan

    Yoshiki Yamaguchi

  2. Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences, Okazaki, Aichi, Japan

    Koichi Kato

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Ji, Y., Woods, R.J. (2018). Quantifying Weak Glycan-Protein Interactions Using a Biolayer Interferometry Competition Assay: Applications to ECL Lectin and X-31 Influenza Hemagglutinin. In: Yamaguchi, Y., Kato, K. (eds) Glycobiophysics. Advances in Experimental Medicine and Biology, vol 1104. Springer, Singapore. https://doi.org/10.1007/978-981-13-2158-0_13

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