Oxygen carrier based on hemoglobin-haptoglobin compound and preparation method of oxygen carrier
A technology of haptoglobin and hemoglobin, which is applied in the field of hemoglobin oxygen carrier-hemoglobin complex oxygen carrier and its preparation, can solve the problems of increasing the mortality rate of subjects, achieve good clinical application prospects, overcome Deficiency, no hypertensive response effect
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Embodiment 1
[0036] Embodiment 1 The present invention is based on the preparation of hemoglobin-haptoglobin complex oxygen carrier
[0037] 1. Preparation method
[0038]Take 0.1 μmol of purified human umbilical cord blood hemoglobin (provided by the Institute of Blood Transfusion, Chinese Academy of Medical Sciences), dissolve it in 10 ml of phosphate buffer (pH 7.4), fill it with pure oxygen for 5 minutes, and add 0.1 μmol of haptoglobin (binding The molar ratio of globin to hemoglobin is about 1:1), then, stir and react in the dark at 0-4°C for 30 minutes, and dialyze the reaction solution with a dialysis bag with a molecular weight cut-off of 100kD until the dialysate outside the dialysis bag is in the 400-600nm region until no absorption. After the dialysis is completed, add 1 μmol of ascorbic acid under stirring, stir and react in the dark at 0-4°C for 5 minutes, adjust the pH value to 7.4 with 0.1M bicarbonate solution, freeze-dry, and obtain the oxygen-carrying carrier of the hem...
Embodiment 2
[0043] Example 2 The present invention is based on the preparation of hemoglobin-haptoglobin complex oxygen carrier
[0044] 1. Preparation method
[0045] Take 0.2 μmol of purified human umbilical cord blood hemoglobin (provided by the Institute of Blood Transfusion, Chinese Academy of Medical Sciences), dissolve it in 10ml of phosphate buffer (pH7.4), fill it with mixed gas (95% oxygen + 5% carbon dioxide) for 5 minutes, and stir Add 0.1 μmol of haptoglobin (the molar ratio of haptoglobin to hemoglobin is about 1:2), then stir and react in the dark at 0-4°C for 5 minutes, and the reaction solution is dialyzed with a dialysis bag with a molecular weight cut-off of 100kD until it is out of the dialysis bag The dialysate has no absorption in the 400-600nm region. After the dialysis is completed, add 0.5 μmol of superoxide dismutase and glutathione under stirring, and add 1.2 μmol of pyridoxal phosphate (the molar ratio of pyridoxal phosphate: hemoglobin is about 6:1), 0~4℃ St...
Embodiment 3
[0050] Example 3 The present invention is based on the preparation of hemoglobin-haptoglobin complex oxygen carrier
[0051] 1. Preparation method
[0052] Take 0.5 μmol of polymerized human cord blood hemoglobin (provided by the Institute of Blood Transfusion, Chinese Academy of Medical Sciences), dissolve it in 10 ml of phosphate buffer (pH 7.4), fill it with a mixed gas (95% oxygen + 5% carbon dioxide) for 10 minutes, and stir Add 0.1 μmol of haptoglobin (the molar ratio of haptoglobin to hemoglobin is about 1:5), then stir and react in the dark at 0-4°C for 15 minutes, and the reaction solution is dialyzed with a dialysis bag with a molecular weight cut-off of 100kD until it is out of the dialysis bag The dialysate has no absorption in the 400-600nm region. After the dialysis is completed, add 2.5 μmol of superoxide dismutase and glutathione under stirring, and add 2 μmol of pyridoxal phosphate (the molar ratio of pyridoxal phosphate: hemoglobin is about 4:1), and keep aw...
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Abstract
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