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Combined characterization of bovine polyhemoglobin microcapsules by UV–Vis absorption spectroscopy and cyclic voltammetry

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Abstract

Polyhemoglobin produced from pure bovine hemoglobin by reaction with PEG bis(N-succynimidil succinate) as a cross-linking agent was encapsulated in gelatin and dehydrated by freeze–drying. Free carboxyhemoglobin and polyhemoglobin microcapsules were characterized by UV–Vis spectroscopy in the absorption range 450–650 nm and cyclic voltammetry in the voltage range from −0.8 to 0.6 mV to evaluate the ability to break the bond with carbon monoxide and to study the carrier’s affinity for oxygen, respectively. SEM used to observe the shape of cross-linked gelatin-polyhemoglobin microparticles showed a regular distribution of globular shapes, with mean size of ~750 nm, which was ascribed to gelatin. Atomic absorption spectroscopy was also performed to detect iron presence in microparticles. Cyclic voltammetry using an Ag–AgCl electrode highlighted characteristic peaks at around −0.6 mV that were attributed to reversible oxygen bonding with iron in oxy-polyhemoglobin structure. These results suggest this technique as a powerful, direct and alternative method to evaluate the extent of hemoglobin oxygenation.

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Acknowledgements

The financial support of Coordination of High-Level Personnel Training (CAPES) (CAPES Scholarship-Proc. No. BEX 3084-14-0) and FAPESP (Proc. No. 2011/21933-0) is gratefully acknowledged. Moreover, Prof. Converti thanks CAPES for the Special Visiting Researcher fellowship within the Program Sciences without Borders (Project No. 2609/2013).

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Authors and Affiliations

  1. Department of Civil, Chemical and Environmental Engineering, University of Genoa, Via Opera Pia 15, 16145, Genoa, Italy

    Marcos Camargo Knirsch, Filippo Dell’Anno, Claudio Larosa & Attilio Converti

  2. Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical Sciences, University of São Paulo, Av. Prof. Lineu Prestes, 580, Bloco 16, São Paulo, SP, CEP 05508-900, Brazil

    Marcos Camargo Knirsch & Bronislaw Polakiewicz

  3. Department of Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy

    Marco Salerno

  4. Department of Surgical Science and Integrate Diagnostic, University of Genoa, Corso Europa 30, 16145, Genoa, Italy

    Roberto Eggenhöffner

  5. INBB, Viale delle Medaglie D’Oro 305, 00136, Rome, Italy

    Roberto Eggenhöffner

Authors
  1. Marcos Camargo Knirsch
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  2. Filippo Dell’Anno
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  3. Marco Salerno
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  4. Claudio Larosa
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  5. Bronislaw Polakiewicz
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  6. Roberto Eggenhöffner
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  7. Attilio Converti
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Correspondence to Attilio Converti.

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Knirsch, M.C., Dell’Anno, F., Salerno, M. et al. Combined characterization of bovine polyhemoglobin microcapsules by UV–Vis absorption spectroscopy and cyclic voltammetry. Bioprocess Biosyst Eng 40, 431–438 (2017). https://doi.org/10.1007/s00449-016-1711-9

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  • DOI: https://doi.org/10.1007/s00449-016-1711-9

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