Process for preparing cross-linked haematoglobin as substitute of red blood cell
A technology of hemoglobin and substitutes, applied in the field of cross-linked hemoglobin, which can solve the problems of high blood pressure, failure to achieve effect, short half-life, etc.
Examples
Embodiment 1
[0022] The pyridoxal hemoglobin solution modified with PLP by pyridoxal 5-phosphate in a conventional manner is used as raw material, the hemoglobin concentration is 11.26% (w / v), and the methemoglobin content is 3.59% (w / w). In an ice bath at 4°C, adjust the pH to 6.70 with 10% vitamin C solution, at this time the concentration of hemoglobin is 10.6%, and the volume is about 755 ml. Feed wet high-purity N 2 For 0.5-1 hour, the hemoglobin is converted into deoxygenated hemoglobin, 0.1% glutathione is added to the solution as an antioxidant, and lysine is pre-added with 5 times the molar amount of hemoglobin. Under sufficient and gentle agitation, add 1% (w / v) glutaraldehyde solution through the sand core sampling device with a constant flow pump at a rate of 0.1 ml / min·100 ml, and the glutaraldehyde and hemoglobin molecules The molar ratio is 13:1, adding in two times, adding 11 parts for the first time, after reacting for ~0.5 hours, then adding the remaining amount of glut...
Embodiment 2
[0024] The operation mode of the cross-linking reaction is the same as the above example. Wherein, the hemoglobin solution is 650 milliliters of the hemoglobin solution that has been purified but not modified by PLP, the hemoglobin concentration is 12%, and the pH is adjusted to 6.70 with vitamin C. In a 4°C ice bath with humidified N 2 Flow deoxygenation for 0.5-1 hour. Add 2% vitamin C or glutathione before the cross-linking reaction, and glycine or ethanolamine with 5 times the molar weight of hemoglobin as cross-linking competition inhibitors, add 1% glutaraldehyde after deoxygenation treatment, and the addition amount is hemoglobin 15 times the molar weight, the addition rate was 0.2 ml / min·100 ml, and the stirring was continued. After 4 hours, use lysine with 11 times the molar amount of hemoglobin as a terminator, and react for 2 hours. Then 59 ml of 1.56% sodium borohydride solution was added for reduction reaction for 4 hours, conventional filtration and ultrafiltr...
Embodiment 3
[0026] Operation process is with embodiment 1.
[0027] The PLP-modified pyridoxal hemoglobin solution is used as a raw material, the hemoglobin concentration is 8%, the volume is 600 ml, and the pH is adjusted to 6.40 with vitamin C. After pre-adding the arginine solution which is 2 times the molar weight of hemoglobin, add 52.5 milliliters of glutaraldehyde with a concentration of 1%, which is 7 times the molar weight of hemoglobin. The adding speed is 0.3 ml / min·100 ml, and the reaction is 30 minutes. Then, adjust the pH=6.90 with 20% disodium hydrogen phosphate, and after pre-adding 4 times the amount of arginine solution, add 26 milliliters of 1% glutaraldehyde 3.5 times the amount, and the adding speed is the same as before. After the second reaction, adjust the pH=7.40 with the same disodium hydrogen phosphate, and then add 4 times the amount of arginine solution, then add the same glutaraldehyde solution of 3.5 times the amount for the third time, and the speed of add...
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Abstract
Description
Claims
Application Information
- IPC
- A61P7/06; C07K14/805
- Inventors
- 杨成民; 王红