title: Fluorescent molecularly imprinted polymers as plastic antibodies for selective labeling and imaging of hyaluronan and sialic acid on fixed and living cells.
abstract: Altered glycosylation levels or distribution of sialic acids (SA) or hyaluronan in animal cells are indicators of pathological conditions like infection or malignancy. We applied fluorescently-labeled molecularly imprinted polymer (MIP) particles for bioimaging of fixed and living human keratinocytes, to localize hyaluronan and sialylation sites. MIPs were prepared with the templates D-glucuronic acid (GlcA), a substructure of hyaluronan, and N-acetylneuraminic acid (NANA), the most common member of SA. Both MIPs were found to be highly selective towards their target monosaccharides, as no cross-reactivity was observed with other sugars like N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, D-glucose and D-galactose, present on the cell surface. The dye rhodamine and two InP/ZnS quantum dots (QDs) emitting in the green and in the red regions were used as fluorescent probes. Rhodamine-MIPGlcA and rhodamine-MIPNANA were synthesized as monodispersed 400nm sized particles and were found to bind selectively their targets located in the extracellular region, as imaged by epifluorescence and confocal microscopy. In contrast, when MIP-GlcA and MIP-NANA particles with a smaller size (125nm) were used, the MIPs being synthesized as thin shells around green and red emitting QDs respectively, it was possible to stain the intracellular and pericellular regions as well. In addition, simultaneous dual-color imaging with the two different colored QDs-MIPs was demonstrated. Importantly, the MIPs were not cytotoxic and did not affect cell viability; neither was the cells morphology affected as demonstrated by live cell imaging. These synthetic receptors could offer a new and promising imaging tool to monitor disease progression.
authors.affiliation: Sorbonne Universités, Université de Technologie de Compiègne, CNRS Enzyme and Cell Engineering Laboratory, Rue Roger Couttolenc, CS 60319, Compiègne Cedex, 60203 France. Electronic address: maria.panagiotopoulou@utc.fr.
authors.forename: Maria
authors.initials: M
authors.lastname: Panagiotopoulou
authors.affiliation: Sorbonne Universités, Université de Technologie de Compiègne, CNRS Enzyme and Cell Engineering Laboratory, Rue Roger Couttolenc, CS 60319, Compiègne Cedex, 60203 France. Electronic address: stephanie.kunath@web.de.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Kunath
authors.affiliation: Sorbonne Universités, Université de Technologie de Compiègne, CNRS Enzyme and Cell Engineering Laboratory, Rue Roger Couttolenc, CS 60319, Compiègne Cedex, 60203 France. Electronic address: paulina.rangel@utc.fr.
authors.forename: Paulina Ximena
authors.initials: PX
authors.lastname: Medina-Rangel
authors.affiliation: Sorbonne Universités, Université de Technologie de Compiègne, CNRS Enzyme and Cell Engineering Laboratory, Rue Roger Couttolenc, CS 60319, Compiègne Cedex, 60203 France. Electronic address: karsten.haupt@utc.fr.
authors.forename: Karsten
authors.initials: K
authors.lastname: Haupt
authors.affiliation: Sorbonne Universités, Université de Technologie de Compiègne, CNRS Enzyme and Cell Engineering Laboratory, Rue Roger Couttolenc, CS 60319, Compiègne Cedex, 60203 France. Electronic address: jeanne.tse-sum-bui@utc.fr.
authors.forename: Bernadette
authors.initials: B
authors.lastname: Tse Sum Bui
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D002460:Cell Line; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis*; D008856:Microscopy, Fluorescence / Q000379:methods; D054802:Molecular Imprinting / Q000379:methods; D019158:N-Acetylneuraminic Acid / Q000032:analysis*; D061848:Optical Imaging / Q000379:methods*; D011108:Polymers / Q000737:chemistry*; D045663:Quantum Dots / Q000737:chemistry*; D012235:Rhodamines / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D011108:Polymers; D012235:Rhodamines; D006820:Hyaluronic Acid; D019158:N-Acetylneuraminic Acid
keywords: Cell imaging; Glycosylation; Hyaluronan; Molecularly imprinted polymers; Quantum dots; Sialic acid