title: Hybrid gravitational field-flow fractionation using immunofunctionalized walls for integrated bioanalytical devices.
abstract: In this work, the biospecific recognition antigen-antibody reaction was implemented in gravitational field-flow fractionation (GrFFF), a flow-assisted separation technique for micron-sized particles, in order to realize a hybrid immunomodulated GrFFF system in which two different principles are combined to achieve highly versatile fractionation. Micron-sized polystyrene beads coated with horseradish peroxidase (HRP) were used as a model sample, and anti-HRP antibodies were immobilized on the accumulation wall of the GrFFF channel. Ultrasensitive chemiluminescence imaging was employed to visualize the beads during elution and to optimize experimental conditions. The same principle was then applied to real biological samples composed by Yersinia enterocolitica and Escherichia coli cells. Results show the possibility to modify the elution of selected sample components and even to retain them into the channel. The hybrid immunomodulated GrFFF system is a step towards the development of a module that could be integrated in a lab-on-a-chip-based point-of-care testing device which includes sample pre-analytical cleanup and analysis.
authors.affiliation: Department of Chemistry G. Ciamician, University of Bologna, Via Selmi 2, 40126, Bologna, Italy. barbara.roda@unibo.it
authors.forename: Barbara
authors.initials: B
authors.lastname: Roda
authors.forename: Sonia
authors.initials: S
authors.lastname: Casolari
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Reschiglian
authors.forename: Mara
authors.initials: M
authors.lastname: Mirasoli
authors.forename: Patrizia
authors.initials: P
authors.lastname: Simoni
authors.forename: Aldo
authors.initials: A
authors.lastname: Roda
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology; D031521:Fractionation, Field Flow / Q000379:methods*; D006735:Horseradish Peroxidase / Q000032:analysis* / Q000276:immunology; D049449:Luminescence; D011137:Polystyrenes / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013499:Surface Properties; D013997:Time Factors; D015008:Yersinia enterocolitica / Q000166:cytology / Q000201:enzymology
chemical_list: D000911:Antibodies, Monoclonal; D011137:Polystyrenes; D006735:Horseradish Peroxidase