MicrogelsCopyright: © L. & F. Biomaterialien
Development of Functionalized Microgels for Specific Binding of Bacterial Enterotoxins
Clostridium difficile (C. difficile) is a toxin-producing bacterium that can colonize the human intestine under given conditions. The released enterotoxins TcdA and TcdB bind to epithelial cells by lectin-glycan interactions and are internalized via endocytosis. As a consequence, toxin-induced inhibition of essential signal transduction pathways destroys the colon epithelium. In this project within the CRC 985 "Microgels", a neo-glycoprotein glycan library identified efficient multivalent TcdA binders. the Lewisy-Lewisx-epitope was found to bind both the receptor domain and the holotoxin. The respective multivalent BSA neo-glycoprotein scavenges TcdA in vitro and protects colorectal HT29 cells from TcdA-induced cell damage (Heine et al. 2019).
The concept of multivalent presentation of glycan epitopes for specific lectin binding and bacterial toxin scavenging was transferred to microgels for the first time (Jans et al. 2017; Boesfeld et al. 2019). GM1a-functionalized microgels scavenge the cholera toxin in vitro and protect colorectal HT29 cells from CT-induced cell damage.
Multi-functional microgels targeted towards inflammatory bowel disease (IBD)
The concept of microgels as scavengers of inflammatory mediators, imaging tools, as well as specific binders to cell surface glycan epitopes is further developed. Functionalization of microgels with galectins and adhesion peptides will target the inflamed tissue thereby building a protective microgel layer to prevent secondary bacterial infection. Microgels with scavenging and imaging units will capture the inflammation ligand TNFα and localize at the sites of inflammation. The ultimate goal is to generate therapeutic, diagnostic, and theranostic microgels.
Publications from the C3 project within the CRC 985 “Microgels”:
P. Bojarová, R. R. Rosencrantz, L. Elling, V. Křen (2013). Enzymatic glycosylation of multivalent scaffolds. Chem. Soc. Rev. 42, 4774-4797. https://doi.org/DOI:10.1039/c2cs35395d
R. R. Rosencrantz, B. Lange, L. Elling (2014). Enzymatic Cascade Reactions for the Synthesis of Glycoconjugates in: Riva, S., Fessner, W.-D. (Eds.), Cascade Biocatalysis - Integrating Stereoselective and Environmentally Friendly Reactions. Wiley-VCH, Weinheim, pp. 133-160. https://doi.org/10.1002/9783527682492.ch6
J. Lazar, H. Park, R. R. Rosencrantz, A. Böker, L. Elling, U. Schnakenberg (2015). Evaluating the Thickness of Multivalent Glycopolymer Brushes for Lectin Binding. Macromol. Rapid Commun. 36, 1472-1478. https://doi.org/10.1002/marc.201500118.
H. Park, R. R. Rosencrantz, L. Elling, A. Böker (2015). Glycopolymer Brushes for Specific Lectin Binding by Controlled Multivalent Presentation of N-Acetyllactosamine Glycan Oligomers. Macromol. Rapid Commun. 36, 45-54. https://doi.org/10.1002/marc.201400453.
J. Lazar, R. R. Rosencrantz, L. Elling, U. Schnakenberg (2016). Simultaneous Electrochemical Impedance Spectroscopy and Localized Surface Plasmon Resonance in a Microfluidic Chip: New Insights into the Spatial Origin of the Signal. Anal. Chem. 88, 9590-9596. https://doi.org/10.1021/acs.analchem.6b02307.
H. Park, S. Walta, R. R. Rosencrantz, A. Körner, C. Schulte, L. Elling, W. Richtering, A. Böker (2016). Micelles from self-assembled double-hydrophilic PHEMA-glycopolymer-diblock copolymers as multivalent scaffolds for lectin binding. Polymer Chem. 7, 878-886. https://doi.org/10.1039/C5PY00797F.
R. R. Rosencrantz, V. H. Nguyen, H. Park, C. Schulte, A. Böker, U. Schnakenberg, L. Elling (2016). Lectin binding studies on a glycopolymer brush flow-through biosensor by localized surface plasmon resonance. Anal. Bioanal. Chem. 408, 5633-5640. https://doi.org/10.1007/s00216-016-9667-9
A. Jans, R. R. Rosencrantz, A. D. Mandić, N. Anwar, S. Boesveld, C. Trautwein, M. Moeller, G. Sellge, L. Elling, A. J. C. Kuehne (2017). Glycan-Functionalized Microgels for Scavenging and Specific Binding of Lectins. Biomacromolecules 18, 1460-1465. https://doi.org/10.1021/acs.biomac.6b01754
S. Boesveld, A. Jans, D. Rommel, M. Bartneck, M. Moeller, L. Elling, C. Trautwein, P. Strnad, A. J. C. Kuehne (2019). Microgels Sopping Up Toxins-GM1a-Functionalized Microgels as Scavengers for Cholera Toxin. ACS Appl. Mat. Interfaces 11, 25017-25023. https://doi.org/10.1021/acsami.9b06413.
V. Heine, S. Boesveld, H. Pelantová, V. Křen, C. Trautwein, P. Strnad, L. Elling (2019). Identifying Efficient Clostridium difficile Toxin A Binders with a Multivalent Neo-Glycoprotein Glycan Library. Bioconj. Chem. 30, 2373-2383. https://doi.org/10.1021/acs.bioconjchem.9b00486.
V. Heine, C. Dey, P. Bojarová, V. Křen, L. Elling (2022). Methods of in vitro study of galectin-glycomaterial interaction. Biotechnol. Adv. 58, 107928. https://doi.org/10.1016/j.biotechadv.2022.107928.
V. Heine, H. Pelantová, P. Bojarová, V. Křen, L. Elling (2022). Targeted Fucosylation of Glycans with Engineered Bacterial Fucosyltransferase Variants. ChemCatChem 14, e202200037. https://doi.org/10.1002/cctc.202200037.