Extraction of superoxide dismutase, catalase, and carbonic anhydrase from stroma-free red blood cell hemolysate for the preparation of the nanobiotechnological complex of polyhemoglobin–superoxide dismutase–catalase–carbonic anhydrase

Abstract

We report a novel method to simultaneously extract superoxide dismutase (SOD), catalase (CAT), and carbonic anhydrase (CA) from the same sample of red blood cells (RBCs). This avoids the need to use expensive commercial enzymes, thus enabling a cost-effective process for large-scale production of a nanobiotechnological polyHb–SOD–CAT–CA complex, with enhancement of all three red blood cell functions. An optimal concentration of phosphate buffer for ethanol–chloroform treatment results in good recovery of CAT, SOD, and CA after extraction. Different concentrations of the enzymes can be used to enhance the activity of polyHb–SOD–CAT–CA to 2, 4, or 6 times that of RBC.

Keywords::

• antioxidant
• blood substitutes
• carbonic anhydrase
• catalase
• enzyme extraction
• erythrocytes
• nanobiotechnology
• nanomedicine
• oxygen carrier
• polyhemoglobin
• stroma-free hemolysate
• superoxide dismutase

Acknowledgements

This laboratory is not connected with any commercial organizations. The research support for the study reported here comes from the joint program of the Canadian Blood Service/Canadian Institutes of Health Research that requires the authors to state that: “The opnions expressed in this article are the opnions of the investigators of this paper and not neccssarily those of the Canadian Government nor of the Canada Blood Service”.

Declaration of interest

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