|VAN TASSELL, MAXWELL - University Of Illinois|
|MILLER, MICHAEL - University Of Illinois|
Submitted to: MethodsX
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/20/2014
Publication Date: 11/7/2014
Publication URL: http://handle.nal.usda.gov/10113/60465
Citation: Van Tassell, M.L., Price, N.P.J., Miller, M.J. 2014. Glycan-specific whole cell affinity chromatography: A versatile microbial adhesion platform. MethodsX. 1:244-250.
Interpretive Summary: Bacteria and other microbes in the human gut often bind to the cell walls of the colon by the use of fimbrial adhesions. These adhesions bind to complex carbohydrates on the surface of gut epithelium cells, and potentially could be selectively competed off by other soluble carbohydrates. These beneficial carbohydrates might for example be provided in the diet. We have developed carbohydrate-modified resins that capture gut bacteria based on their selective adhesion. The resins are straightforward to prepare and are based on ARS-developed “C-glycoside” chemistry. Using these new resins, we show 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 their fimbrial adhesions. The new technique might be used for developing anti-adhesive therapeutics, or for testing the beneficial effects of carbohydrates in the diet
Technical 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.