Abstract
A new protocol is described for derivatization of hemoglobin with polyethyleneglycol (PEG) via reaction of the unmodified native hemoglobin with an activated amine-reacting polyethylene glycol derivative which, unlike protocols previously described, leads to formation of a peptide bond between hemoglobin and PEG. Dioxygen binding and peroxide reactivities of the derivatized hemoglobin are examined, and found to be within reasonable limits, with the particular observation that, unlike with a few other derivatization protocols, the dioxygen affinity is slightly lower than that of native Hb. In cell culture tests (human umbilical vein epithelial cells, HUVEC), the derivatization protocol induces no toxic effect. These results show promise towards applicability for production of hemoglobin-based blood substitutes.
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Abbreviations
- PEG:
-
Polyethyleneglycol
- Hb:
-
Hemoglobin
- HUVEC:
-
Human umbilical vein epithelial cells
- PEG-Hb:
-
Hemoglobin derivatized with polyethyleneglycol
- PBS:
-
Phosphate buffer saline
- TMS:
-
(methyl-PEG12)3-PEG4-N-hydroxysuccinimide ester
- DMSO:
-
Dimethyl sulfoxide
- MTT:
-
3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide
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Acknowledgments
The work shown here has been supported by the Romanian Ministry for Education and Research (grants PNII ID565/2007 and PCCE 140/2008) and by PhD scholarships to FD and AM (Contract POSDRU/88/1.5/S/60185—“Innovative doctoral studies in a knowledge based society”).
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Zolog, O., Mot, A., Deac, F. et al. A New Polyethyleneglycol-Derivatized Hemoglobin Derivative with Decreased Oxygen Affinity and Limited Toxicity. Protein J 30, 27–31 (2011). https://doi.org/10.1007/s10930-010-9298-5
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DOI: https://doi.org/10.1007/s10930-010-9298-5