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Glycan-specific whole cell affinity chromatography: A versatile microbial adhesion platform

Author:
Maxwell L. Van Tassell, Neil P. J. Price, Michael J. Miller
Source:
MethodsX 2014 v.1 pp. 244-250
ISSN:
2215-0161
Subject:
Escherichia coli, affinity chromatography, bacterial adhesion, mannose, microorganisms, polysaccharides, resins
Abstract:
We have constructed a C-glycoside ketohydrazide affinity chromatography resin that interacts with viable whole-cell microbial populations with biologically appropriate stereo-specificity in a carbohydrate-defined manner. It readily allows for the quantification, selection, and manipulation of target organisms based on adhesion to glycan ligands. Construction of the resin does not involve lengthy procedures, highly reactive reagents, or sophisticated equipment, making it simple to construct even in settings with little expertise in chemistry or biotechnology. We demonstrate that a laboratory strain of Escherichia coli adheres to immobilized mannose in a glycan-specific fashion and that populations of E. coli can be separated based on the regulatory state of fimbrial adhesions. Such a platform could be used to investigate adhesive mechanisms of microbial organisms for developing anti-adhesive therapeutics and elaborating the dynamics of competitive inhibition by commensal and probiotic organisms.
Agid:
60465
Handle:
10113/60465