Next Article in Journal
Synthesis and Biological Evaluation of a New Polymeric Conjugate and Nanocarrier with Osteotropic Properties
Next Article in Special Issue
Replacing the Transfusion of 1–2 Units of Blood with Plasma Expanders that Increase Oxygen Delivery Capacity: Evidence from Experimental Studies
Previous Article in Journal / Special Issue
Biophysical Properties of Lumbricus terrestris Erythrocruorin and Its Potential Use as a Red Blood Cell Substitute
J. Funct. Biomater. 2012, 3(1), 61-78; doi:10.3390/jfb3010061
Article

Monodisperse 130 kDa and 260 kDa Recombinant Human Hemoglobin Polymers as Scaffolds for Protein Engineering of Hemoglobin-Based Oxygen Carriers

1,2,†
,
1,2,‡
,
2, 2,§
,
2,||
,
1,2
 and
2,¶,*
Received: 3 November 2011 / Revised: 29 December 2011 / Accepted: 5 January 2012 / Published: 13 January 2012
(This article belongs to the Special Issue Blood Substitutes)
View Full-Text   |   Download PDF [2070 KB, 16 January 2012; original version 13 January 2012]   |   Browse Figures

Abstract

A recombinant 130 kDa dihemoglobin which is made up of a single-chain tetra-α globin and four β globins has been expressed as a soluble protein in E. coli. The sequence of the single chain tetra-α is: αI-Gly-αII-(SerGlyGly)5Ser-αIII-Gly-αIV. This dihemoglobin has been purified and characterized in vitro by size exclusion chromatography, electrospray mass spectroscopy, equilibrium oxygen binding, and analytical ultracentrifugation. The observed values of P50 and nmax for the dihemoglobin are slightly lower than those observed for the recombinant hemoglobin rHb1.1 (a “monohemoglobin” comprised of two β globins and an αI-Gly-αII diα-globin chain). Titration of the deoxy form of dihemoglobin with CO shows that all eight heme centers bind ligand. In vivo, dihemoglobin showed increased circulating halflife and a reduced pressor response in conscious rats when compared to rHb1.1. These observations suggest that dihemoglobin is an oxygen carrying molecule with desirable in vivo properties and provides a platform for an isooncotic hemoglobin solution derived solely from a recombinant source. A 260 kDa tetrahemoglobin has also been produced by chemical crosslinking of a dihemoglobin that contains a Lys16Cys mutation in the C-terminal α-globin subunit. Tetrahemoglobin also shows reduced vasoactivity in conscious rats that is comparable to that observed for dihemoglobin.
Keywords: polyhemoglobin; dihemoglobin; tetrahemoglobin; hemoglobin engineering; extravasation; pressor response polyhemoglobin; dihemoglobin; tetrahemoglobin; hemoglobin engineering; extravasation; pressor response
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Share & Cite This Article

Further Mendeley | CiteULike
Export to BibTeX |
EndNote
MDPI and ACS Style

Marquardt, D.A.; Doyle, M.P.; Davidson, J.S.; Epp, J.K.; Aitken, J.F.; Lemon, D.D.; Anthony-Cahill, S.J. Monodisperse 130 kDa and 260 kDa Recombinant Human Hemoglobin Polymers as Scaffolds for Protein Engineering of Hemoglobin-Based Oxygen Carriers. J. Funct. Biomater. 2012, 3, 61-78.

View more citation formats

Related Articles

Article Metrics

Comments

Cited By

[Return to top]
J. Funct. Biomater. EISSN 2079-4983 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert