Method for preparing red blood cell substitutes using polyaldehyde compounds

Abstract

A method for preparing red blood cell substitutes with polyaldehyde-based compounds. In the reaction of polymerizing hemoglobin, the present invention reduces the concentration of the reaction substrate, and quickly adds the polyaldehyde-based compound solution to the hemoglobin solution for reaction; after the reaction is terminated, the obtained molecular weight is 100 The hemoglobin polymer between ~1500kDa; Compared with the prior art, the present invention has: 1) fast addition of polyaldehyde compounds, which greatly shortens the polymerization process, can shorten the production cycle and reduce costs; The compound maintains a high concentration for a relatively long period of time and has the function of inactivating the virus; ③More than 90% of the hemoglobin molecules are cross-linked within the molecule, making the structure of hemoglobin stable; ④Higher pH value can improve the reaction solution. The concentration of free polyaldehyde compounds in the medium can reduce the content of uncrosslinked hemoglobin between molecules, which can reduce toxicity; ⑤The average molecular weight of hemoglobin after polymerization increases significantly, and its half-life can reach 18-25h, which is more conducive to clinical application.

Description

technical field [0001] The invention belongs to the technical field of biological products, and in particular relates to a preparation method of a red blood cell substitute (blood substitute) with hemoglobin as a substrate. Background technique [0002] Blood transfusion is a very important medical method in clinical operations, disaster relief, terrorist attacks and field rescue. At present, people mainly rely on people to donate blood to meet the needs. Due to the complex blood type of human beings, matching must be carried out during blood transfusion, and the storage period of blood is short, transportation is inconvenient, the source is limited, and there are potential risks of virus infection such as AIDS and hepatitis B. Limited, especially heavy use in emergencies. In recent years, with the continuous increase of blood demand, it is difficult to meet the increasing demand only by human blood donation. Therefore, the development of safe and effective blood substitute...

AI Technical Summary

Problems solved by technology

[0004] However, the direct use of stroma-free hemoglobin in the human body has the following problems: first, it will produce strong toxicity, mainly manifested as hematuria, increased blood pressure, abdominal pain, renal failure and even death. The main reason is that the tetrameric hemoglobin molecule ( 64kDa) is easily dissociated into dimers with a smaller molecular weight (32kDa), and these small molecules are easy to seep out from the intercellular space of the vascular endothelium, causing the above-mentioned toxic and side effects; second, the residence time of stroma-free hemoglobin in the body is very short , the half-life is only about 4h, which cannot meet the requirements of clinical use, because hemoglobin is a protein composed of four subunits (molecular weight is 64kDa), and the four subunits are respectively composed of two pairs of αβ subunits. It is held together by ionic bonds, hydrophobic interactions and van der Waals forces rather than covalent bonds, so it is easy to dissociate into αβ subunits of dimers, causing the above toxicity

In all current technologies for preparing red blood cell substitutes, the addition of polyaldehyde-based compounds is done dropwise, which makes the polymerization process take a long time. For example, U.S. Patent No. 5,895,810 describes a method for cross-linking hemoglobin with glutaraldehyde , the process of its polymerization reaction exceeds 6 hours, which is not conducive to industrial mass production