TY - JOUR
T1 - Glycan-specific whole cell affinity chromatography
T2 - A versatile microbial adhesion platform
AU - Van Tassell, Maxwell L.
AU - Price, Neil P.J.
AU - Miller, Michael J.
N1 - Funding Information:
The authors would like to thank Trina Hartman for technical assistance and the Miller lab group for critical review of the manuscript. This work was supported by funds awarded to M.J.M. from the United States Department of Agriculture, National Institute of Food and Agriculture (Award Number 2011-67015-30772). Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture. USDA is an equal opportunity provider and employer.
Publisher Copyright:
© 2014 The Authors. Published by Elsevier B.V.
PY - 2014
Y1 - 2014
N2 - (Figure Presented) We have sought a universal platform for elucidating and exploiting specificity of glycan-mediated adhesion by potentially uncharacterized microorganisms. Several techniques exist to explore microbial interactions with carbohydrate structures. Many are unsuitable for investigating specific mechanisms or uncharacterized organisms, requiring pure cultures, labeling techniques, expensive equipment, or other limitations such as questionable stability, stereospecificity, or scalability. We have adapted an affinity chromatography resin as a model to overcome these drawbacks, among others. It readily allows for the quantification, selection, and manipulation of target organisms based on interactions with glycan ligands. To maximize its utility as a selective screening method, we have constructed the tool such that it: • Promotes whole-cell interactions using viable, unaltered cells. • Provides robust spatial interactions with target glycans, presented with controlled stereo-specificity, for high affinity/avidity interactions that reflect a complex in vivo matrix. • Has the ability to utilize any reducing glycan, is quick, efficient, safe, and affordable to construct, and is scalable and reusable for multiple applications.
AB - (Figure Presented) We have sought a universal platform for elucidating and exploiting specificity of glycan-mediated adhesion by potentially uncharacterized microorganisms. Several techniques exist to explore microbial interactions with carbohydrate structures. Many are unsuitable for investigating specific mechanisms or uncharacterized organisms, requiring pure cultures, labeling techniques, expensive equipment, or other limitations such as questionable stability, stereospecificity, or scalability. We have adapted an affinity chromatography resin as a model to overcome these drawbacks, among others. It readily allows for the quantification, selection, and manipulation of target organisms based on interactions with glycan ligands. To maximize its utility as a selective screening method, we have constructed the tool such that it: • Promotes whole-cell interactions using viable, unaltered cells. • Provides robust spatial interactions with target glycans, presented with controlled stereo-specificity, for high affinity/avidity interactions that reflect a complex in vivo matrix. • Has the ability to utilize any reducing glycan, is quick, efficient, safe, and affordable to construct, and is scalable and reusable for multiple applications.
KW - Affinity chromatography
KW - Bacterial adhesion
KW - Campylobacter jejuni
KW - Carbohydrate modification
KW - Escherichia coli
KW - Lectins
KW - Screening
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U2 - 10.1016/j.mex.2014.10.005
DO - 10.1016/j.mex.2014.10.005
M3 - Article
C2 - 26150959
AN - SCOPUS:84949129727
SN - 2215-0161
VL - 1
SP - 244
EP - 250
JO - MethodsX
JF - MethodsX
ER -