Magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes.

A new method for hemoglobin (Hb) deoxygenation, in suspension or within red blood cells (RBCs) is described using the commercial enzyme product, EC-Oxyrase®. The enzymatic deoxygenation method has several advantages over established deoxygenation methodologies, such as avoiding side reactions that p...

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Autores principales:	Mitchell R H Weigand, Jenifer Gómez-Pastora, James Kim, Matthew T Kurek, Richard J Hickey, David C Irwin, Paul W Buehler, Maciej Zborowski, Andre F Palmer, Jeffrey J Chalmers
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/43431974dcc5428b83f6d54eb72192ce
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spelling	oai:doaj.org-article:43431974dcc5428b83f6d54eb72192ce2021-12-02T20:08:31ZMagnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes.1932-620310.1371/journal.pone.0257061https://doaj.org/article/43431974dcc5428b83f6d54eb72192ce2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0257061https://doaj.org/toc/1932-6203A new method for hemoglobin (Hb) deoxygenation, in suspension or within red blood cells (RBCs) is described using the commercial enzyme product, EC-Oxyrase®. The enzymatic deoxygenation method has several advantages over established deoxygenation methodologies, such as avoiding side reactions that produce methemoglobin (metHb), thus eliminating the need for an inert deoxygenation gas and airtight vessel, and facilitates easy re-oxygenation of Hb/RBCs by washing with a buffer that contains dissolved oxygen (DO). The UV-visible spectra of deoxyHb and metHb purified from human RBCs using three different preparation methods (sodium dithionite [to produce deoxyHb], sodium nitrite [to produce metHb], and EC-Oxyrase® [to produce deoxyHb]) show the high purity of deoxyHb prepared using EC-Oxyrase® (with little to no metHb or hemichrome production from side reactions). The oxyHb deoxygenation time course of EC-Oxyrase® follows first order reaction kinetics. The paramagnetic characteristics of intracellular Hb in RBCs were compared using Cell Tracking Velocimetry (CTV) for healthy and sickle cell disease (SCD) donors and oxygen equilibrium curves show that the function of healthy RBCs is unchanged after EC-Oxyrase® treatment. The results confirm that this enzymatic approach to deoxygenation produces pure deoxyHb, can be re-oxygenated easily, prepared aerobically and has similar paramagnetic mobility to existing methods of producing deoxyHb and metHb.Mitchell R H WeigandJenifer Gómez-PastoraJames KimMatthew T KurekRichard J HickeyDavid C IrwinPaul W BuehlerMaciej ZborowskiAndre F PalmerJeffrey J ChalmersPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 9, p e0257061 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mitchell R H Weigand Jenifer Gómez-Pastora James Kim Matthew T Kurek Richard J Hickey David C Irwin Paul W Buehler Maciej Zborowski Andre F Palmer Jeffrey J Chalmers Magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes.
description	A new method for hemoglobin (Hb) deoxygenation, in suspension or within red blood cells (RBCs) is described using the commercial enzyme product, EC-Oxyrase®. The enzymatic deoxygenation method has several advantages over established deoxygenation methodologies, such as avoiding side reactions that produce methemoglobin (metHb), thus eliminating the need for an inert deoxygenation gas and airtight vessel, and facilitates easy re-oxygenation of Hb/RBCs by washing with a buffer that contains dissolved oxygen (DO). The UV-visible spectra of deoxyHb and metHb purified from human RBCs using three different preparation methods (sodium dithionite [to produce deoxyHb], sodium nitrite [to produce metHb], and EC-Oxyrase® [to produce deoxyHb]) show the high purity of deoxyHb prepared using EC-Oxyrase® (with little to no metHb or hemichrome production from side reactions). The oxyHb deoxygenation time course of EC-Oxyrase® follows first order reaction kinetics. The paramagnetic characteristics of intracellular Hb in RBCs were compared using Cell Tracking Velocimetry (CTV) for healthy and sickle cell disease (SCD) donors and oxygen equilibrium curves show that the function of healthy RBCs is unchanged after EC-Oxyrase® treatment. The results confirm that this enzymatic approach to deoxygenation produces pure deoxyHb, can be re-oxygenated easily, prepared aerobically and has similar paramagnetic mobility to existing methods of producing deoxyHb and metHb.
format	article
author	Mitchell R H Weigand Jenifer Gómez-Pastora James Kim Matthew T Kurek Richard J Hickey David C Irwin Paul W Buehler Maciej Zborowski Andre F Palmer Jeffrey J Chalmers
author_facet	Mitchell R H Weigand Jenifer Gómez-Pastora James Kim Matthew T Kurek Richard J Hickey David C Irwin Paul W Buehler Maciej Zborowski Andre F Palmer Jeffrey J Chalmers
author_sort	Mitchell R H Weigand
title	Magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes.
title_short	Magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes.
title_full	Magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes.
title_fullStr	Magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes.
title_full_unstemmed	Magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes.
title_sort	magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: chemical versus enzymatic processes.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/43431974dcc5428b83f6d54eb72192ce
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Magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes.

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