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Oxygen binding of modified hemoglobins in solution
D Mauldin1, R L Dalpé, P J Anderson
1Department of Physiology, University of Ottawa, Canada.
Advances in Experimental Medicine and Biology
|January 1, 1989
Summary
Pyridoxal phosphate modification of SFHS using NaCNBH3 enhances oxygen carrying capacity and cooperativity. This method yields superior functional hemoglobin compared to NaBH4, avoiding high methemoglobin levels.
Area of Science:
- Biochemistry
- Biomedical Engineering
- Hemoglobin Research
Background:
- Hemoglobin (Hb) is crucial for oxygen transport.
- Modifications to Hb aim to improve oxygen delivery and stability.
- Pyridoxal phosphate (PLP) is a known Hb modifier.
Purpose of the Study:
- To evaluate the efficacy of SFHS (Stroma-Free Hemoglobin Solution) modified with PLP using different reducing agents.
- To compare the functional properties of SFHS-P prepared with NaCNBH3 versus NaBH4.
Main Methods:
- Covalent incorporation of pyridoxal phosphate into SFHS using sodium cyanoborohydride (NaCNBH3) as the reducing agent.
- Characterization of the modified SFHS-P, including P50, Hill slope, oxygen carrying capacity, oxygen saturation, and methemoglobin levels.
- Comparative analysis with SFHS-P prepared using sodium borohydride (NaBH4).
Main Results:
- SFHS-P modified with NaCNBH3 exhibited P50 values of 21-25 mm Hg and a Hill slope of 2.34, indicating good cooperativity.
- Oxygen carrying capacity was approximately 1.3, with >90% oxygen saturation and <3% methemoglobin.
- SFHS-P prepared with NaBH4 showed high oxygen affinity, elevated methemoglobin, and poor oxygen saturation, with a lowered oxygen carrying capacity.
Conclusions:
- NaCNBH3 is a superior reducing agent for PLP modification of SFHS compared to NaBH4.
- The NaCNBH3 method yields a functional hemoglobin product with desirable oxygen transport characteristics.
- SFHS-P prepared via NaBH4 exhibits significant deficiencies, limiting its potential utility.