title: Development of recombinant hemoglobin-based oxygen carriers.
abstract: CONCLUSIONS
  The worldwide blood shortage has generated a significant demand for alternatives to whole blood and packed red blood cells for use in transfusion therapy. One such alternative involves the use of acellular recombinant hemoglobin (Hb) as an oxygen carrier.
  
  
  BACKGROUND
  Large amounts of recombinant human Hb can be expressed and purified from transgenic Escherichia coli. The physiological suitability of this material can be enhanced using protein-engineering strategies to address specific efficacy and toxicity issues. Mutagenesis of Hb can (i) adjust dioxygen affinity over a 100-fold range, (ii) reduce nitric oxide (NO) scavenging over 30-fold without compromising dioxygen binding, (iii) slow the rate of autooxidation, (iv) slow the rate of hemin loss, (v) impede subunit dissociation, and (vi) diminish irreversible subunit denaturation. Recombinant Hb production is potentially unlimited and readily subjected to current good manufacturing practices, but may be restricted by cost. Acellular Hb-based O(2) carriers have superior shelf-life compared to red blood cells, are universally compatible, and provide an alternative for patients for whom no other alternative blood products are available or acceptable.
  
  
  RESULTS
  Remaining objectives include increasing Hb stability, mitigating iron-catalyzed and iron-centered oxidative reactivity, lowering the rate of hemin loss, and lowering the costs of expression and purification. Although many mutations and chemical modifications have been proposed to address these issues, the precise ensemble of mutations has not yet been identified.
  
  
  CONCLUSIONS
  Future studies are aimed at selecting various combinations of mutations that can reduce NO scavenging, autooxidation, oxidative degradation, and denaturation without compromising O(2) delivery, and then investigating their suitability and safety in vivo.
authors.affiliation: Department of Biochemistry & Cell Biology, Rice University, Houston, TX 77005, USA.
authors.forename: Cornelius L
authors.initials: CL
authors.lastname: Varnado
authors.forename: Todd L
authors.initials: TL
authors.lastname: Mollan
authors.forename: Ivan
authors.initials: I
authors.lastname: Birukou
authors.forename: Bryan J Z
authors.initials: BJ
authors.lastname: Smith
authors.forename: Douglas P
authors.initials: DP
authors.lastname: Henderson
authors.forename: John S
authors.initials: JS
authors.lastname: Olson
mesh_terms: D000818:Animals; D001692:Biological Transport; D001802:Blood Substitutes; D006418:Heme / Q000378:metabolism; D006454:Hemoglobins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D009569:Nitric Oxide / Q000378:metabolism; D010084:Oxidation-Reduction; D010100:Oxygen / Q000097:blood*; D011485:Protein Binding; D055550:Protein Stability; D011994:Recombinant Proteins
chemical_list: D001802:Blood Substitutes; D006454:Hemoglobins; D011994:Recombinant Proteins; D009569:Nitric Oxide; D006418:Heme; D010100:Oxygen