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Hybrid gravitational field-flow fractionation using immunofunctionalized walls for integrated bioanalytical devices

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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.

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Acknowledgments

This work was financially supported by the University of Bologna (“Strategic University Projects” and “RFO—Focused fundamental research projects”) and by the Italian Ministry for Education, University and Research (“PRIN 2007” projects).

The authors are grateful to the Istituto Zooprofilattico “G. Caporale” (Teramo, Italy) for kindly providing the anti-Yersinia-enterocolitica monoclonal antibody.

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Authors and Affiliations

  1. Department of Chemistry “G. Ciamician”, University of Bologna, Via Selmi 2, 40126, Bologna, Italy

    Barbara Roda, Sonia Casolari & Pierluigi Reschiglian

  2. Laboratory of Analytical and Bioanalytical Chemistry, Department of Pharmaceutical Sciences, University of Bologna, Via Belmeloro 6, 40126, Bologna, Italy

    Mara Mirasoli & Aldo Roda

  3. Department of Clinical Medicine, University of Bologna, via Massarenti 9, 40138, Bologna, Italy

    Patrizia Simoni

  4. National Institute of Biostructure and Biosystems, NIBB, Interuniversity Consortium, Rome, Italy

    Barbara Roda, Sonia Casolari, Pierluigi Reschiglian, Mara Mirasoli, Patrizia Simoni & Aldo Roda

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  1. Barbara Roda
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  2. Sonia Casolari
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  3. Pierluigi Reschiglian
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  4. Mara Mirasoli
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  5. Patrizia Simoni
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  6. Aldo Roda
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Correspondence to Barbara Roda.

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Roda, B., Casolari, S., Reschiglian, P. et al. Hybrid gravitational field-flow fractionation using immunofunctionalized walls for integrated bioanalytical devices. Anal Bioanal Chem 394, 953–961 (2009). https://doi.org/10.1007/s00216-009-2714-z

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  • DOI: https://doi.org/10.1007/s00216-009-2714-z

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