PEGylation of αα-Hb using succinimidyl propionic acid PEG 5K: Conjugation chemistry and PEG shell structure dictate respectively the oxygen affinity and resuscitation fluid like properties of PEG αα-Hbs

Abstract

PEGylation of intramolecularly crosslinked Hb has been studied here to overcome the limitation of dissociation of Hb tetramers. New hexa and deca PEGylated low oxygen affinity PEG-ααHbs have been generated. Influence of PEG conjugation chemistry and the PEG shell structure on the functional properties as well as PEGylation induced plasma expander like properties of the protein has been delineated. The results have established that in the design of PEG-Hbs as oxygen therapeutics, the influence of conjugation chemistry and the PEG shell structure on the oxygen affinity of Hb needs to be optimized independently besides optimizing the PEG shell structure for inducing resuscitation fluid like properties.

Keywords::

• blood substitute
• PEGylation
• PEGylated hemoglobin
• PEG shell
• plasma expander
• oxygen affinity

Acknowledgements

We thank Dr Abraham Abuchowski (Prolong Pharmaceuticals) for providing us with the succinimidyl carbamate PEG5K reagent.

Declaration of interest

The authors report no declarations of interest. The authors alone are responsible for the content and writing of the paper.

This research is partially supported by the United States Army Medical Research Acquisition Activity Grant (W81XWH-11-2-0012) to AGT.