Document Detail

Synthesis and characterization of a novel DTPA polymerized hemoglobin based oxygen carrier.
MedLine Citation:
PMID:  16102904     Owner:  NLM     Status:  MEDLINE    
OBJECTIVE: The purpose of this study was to prepare a novel polymerized hemoglobin (Hb) based oxygen carrier (HBOC) designed to minimize Hb induced hypertension, while employing a simple and inexpensive method of preparation. Cyclic-diethylenetriaminepentaacetic acid (DTPA) was used to polymerize stroma free Hb (SF-Hb). METHODS: SF-Hb was isolated from red blood cells and reacted with DTPA at a constant concentration, pH, and duration. Low molar mass fractions (<100 kDa) were removed using ultrafiltration. Reactions and subsequent ultrafiltration steps were determined to be reproducible by analyzing molar mass, colloid osmotic pressure and oxygen affinity. Finally, a model of 50% exchange transfusion (ET) in rats was used to evaluate the blood pressure response to DTPA polymerized SF-Hb (Poly-DTPA-Hb). RESULTS: Poly-DTPA-Hb demonstrated a number averaged molar mass of 128.7 kDa and a weighted average of 223.0 kDa. Oxygen binding equilibrium indicated high oxygen affinity (P50 = 5.1+/-0.01 mm Hg) and little cooperativity (n = 1.4). Poly-DTPA-Hb and a control DTPA polymerized human serum albumin (Poly-DTPA-HSA) unexpectedly caused acute hypotension during the period of ET in rats (mean arterial pressure approximately 45% less than baseline). Hypotension occurring over the period of ET was determined to be mediated by calcium binding to protein associated DTPA. This effect was attenuated by the addition of calcium chloride (CaCl2) to the Poly-DTPA protein preparations. CONCLUSIONS: Cyclic DTPA anhydride can be used to create cross-linked and polymerized hemoglobin, using a simple and inexpensive process. However, the addition of CaCl2 to the preparation appears to be required to prevent calcium chelation and subsequent hypotension during infusion.
Chad R Haney; Paul W Buehler; Anil Gulati
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  1725     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  2005 Oct 
Date Detail:
Created Date:  2005-10-03     Completed Date:  2005-11-28     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  358-69     Citation Subset:  IM    
Department of Bioengineering, The University of Illinois at Chicago, Chicago, IL 60612, USA.
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MeSH Terms
Blood Pressure / drug effects
Blood Substitutes / chemical synthesis*
Chromatography, Gel
Exchange Transfusion, Whole Blood
Hemoglobins / chemistry*
Molecular Weight
Nephelometry and Turbidimetry
Osmotic Pressure
Oxygen / blood
Pentetic Acid / analogs & derivatives*,  chemistry
Rats, Sprague-Dawley
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Grant Support
Reg. No./Substance:
0/Blood Substitutes; 0/Hemoglobins; 0/hemoglobin, stroma free; 23911-26-4/cyclic DTPA anhydride; 67-43-6/Pentetic Acid; 7782-44-7/Oxygen

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