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Modification method of polymeric hemoglobin

A technology of hemoglobin and modification methods, applied in the fields of peptide/protein components, blood diseases, drug combinations, etc.

Inactive Publication Date: 2007-07-04
天津协和生物科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the literature reviewed, the PLP modification of hemoglobin is placed before the polymerization step (pre-modification), and no one has tried to place the modification step after the polymerization step (post-modification).

Method used

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  • Modification method of polymeric hemoglobin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: PLP pre-modification and post-modification

[0022] I will pass the hemoglobin purified from placental blood (or adult peripheral blood) at a low temperature of 0 to 4°C, and inert gas such as high-purity nitrogen without oxygen, with a flow rate of 1 to 5 L per minute, for 15 ~30 minutes, and add conventional bacteriostatic drugs such as penicillin and / or streptomycin in an amount of 100,000 to 1 million units / 1000 ml and mix well; dissolve PLP with 1M Tris-Hcl, the concentration is 2%, PH =7.3-7.4, add according to the molar ratio of hemoglobin 4:1-8:1, continue to react with inert gas such as high-purity nitrogen for 0.5-1.5 hours, add 2% concentration of trough at a weight / volume ratio of 0.1-0.5% As an antioxidant, add 1.56% NaBH at a molar ratio of 20:1-80:1 to hemoglobin after reacting with inert gas such as high-purity nitrogen for 1-2 hours. 4 (dissolved in 10 -3 M sodium hydroxide) terminated, and reacted for 3-6 hours. Then, the hemoglobin solut...

Embodiment 2

[0024] Embodiment 2: the adding order of glutathione:

[0025] I will pass the hemoglobin purified from placental blood (or adult peripheral blood) at a low temperature of 0 to 4°C, and inert gas such as high-purity nitrogen without oxygen, with a flow rate of 1 to 5 L per minute, for 15 ~30 minutes, add conventional antibacterial drugs such as penicillin and / or streptomycin in an amount of 100,000 to 1,000,000 units / 1000 ml and mix well, then pass inert gas such as high-purity nitrogen for 0.5-1 hour according to weight / volume The ratio is 0.1 to 0.5% and 2% glutathione is added as an antioxidant. Dissolve PLP with 1M Tris-HCl at a concentration of 2%, pH=7.3-7.4, add it at a molar ratio of 4:1-8:1 to hemoglobin, and continue to react with inert gas such as high-purity nitrogen for 2-3 hours. Add 1.56% NaBH4 (dissolved in 10 -3 M sodium hydroxide) to terminate the reaction for 3-6 hours. Then the hemoglobin solution is subjected to ultrafiltration treatment in a convention...

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Abstract

The invention relates to a modification method of polymmeric hemoglobin, it is characterized by: preparing purified hemoglobin from natural red blood cells in the condition of 0-4DEG C and nitrogen, first polymerizing them through same type bifunctional reagent, then modifying them with 5'-pyridoxal phosphate. During the modifying process, glutathione should be added after adding PLP 0.5-1.5 hours. The arts can control generation of ferrihemoglobin, but also ensuret the P50 value in appropriate range and reduce production costs greatly.

Description

(1) Technical field: [0001] The invention relates to the field of biomedicine, in particular to a method for modifying polymerized hemoglobin. (two) background technology: [0002] The traditional blood transfusion method with transfusion of whole blood as the main means has played an extremely important role in medical rescue, but at the same time, it also has various shortcomings that are difficult to overcome. Risk of viral cross-infection, strained blood sources, short storage period and special facilities are required. In order to solve the above contradictions, people started research on red blood cell substitutes as early as the end of the 19th century. Red blood cell substitute refers to the general term for various oxygen carriers that can expand blood volume, maintain the balance of osmotic pressure inside and outside blood vessels, and have the function of carrying oxygen to replace human red blood cells. The development of blood substitutes has gone through the...

Claims

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Application Information

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IPC IPC(8): A61K38/42C08H1/00A61P7/08
Inventor 杨成民李凤娟张鸿辉梁伟光李洪英许丽丽李燊王劲峰杨晓明
Owner 天津协和生物科技发展有限公司
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