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Method for chromatographic removal of prions

A prion and exchange chromatography technology, applied in the field of chromatographic separation of prions, can solve problems such as destructive activity and denaturation

Inactive Publication Date: 2006-11-08
HEMOGLOBIN OXYGEN THERAPEUTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The extreme conditions required to remove the infectivity of prions are generally incompatible with methods of preserving the useful activity of biomolecules, resulting in denaturation of many biomolecules and essentially destroying their activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 adopts the purification of bovine hemoglobin by anion exchange chromatography

[0029] Preparation of bovine hemoglobin solution

[0030] The preparation of the bovine hemoglobin solution was performed as described in US Patent Application Serial No. 08 / 473,497. First, a sample of bovine whole blood is collected, mixed with the anticoagulant sodium citrate to form a citrated blood solution, and then analyzed for endotoxin levels. After a sample of the blood solution was collected, it was stored at a temperature of 2° C., and then filtered through a 600-mesh sieve to remove large clots and particles.

[0031] The citrated blood solution was then sequentially passed through 800 μm and 50 μm polypropylene filters to remove large debris from the blood solution.

[0032] The red blood cells are then washed to separate extracellular plasma proteins such as BSA or IgG from the red blood cells. When washing red blood cells, first place the blood solution in a di...

Embodiment 2

[0051] Example 2 Confirmation of the method for removing prions from bovine hemoglobin preparations

[0052] The confirmation of the above-mentioned removal method was carried out in a specially registered facility, and the procedures followed were in compliance with the US Food and Drug Administration's laboratory operation regulations (21CFR 58), the procedures stipulated by the British GLP, and the regulations of the Japanese GLP and OECD laboratory operating principles [U.S. Food and Drug Administration Good Laboratory Practice Regulation (21 CFR 58), the United Kingdom GLP Compliance Programme, the Japanese GLP Standard and the OECD Principles of Good Laboratory Practice].

[0053] Various solutions for evaluating the infectivity of sheep dysentery by in vivo assay are: 1) solution of sheep dysentery causative agent for infecting hemoglobin solution; 2) sample A'; 3) sample A"; 4) sample B' ; 5) Sample B".

[0054] In vivo detection

[0055] The detection of sheep dysen...

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Abstract

A method for removing a prion from a solution comprising the prion and at least one additional biomolecule, comprising directing the solution through an anion-exchange chromatography column under conditions that cause a gradient elution, whereby the prion is separated from at least one of the biomolecules, thereby causing said biomolecule to be collected in an eluate fraction that is distinct from an eluate fraction that includes the prion. In one embodiment, the gradient is a pH gradient, for example, a step gradient. The prion can be a causal agent for a spongiform encephalopathy, such as Creutzfeldt-Jakob disease, Gerstmann-Straussler-Scheinken syndrome, scrapie, or bovine spongiform encephalopathy.

Description

technical field [0001] The present invention relates to a method for removing prions from a solution containing prions and hemoglobin, the method comprising the step of passing the solution through an anion exchange chromatographic column under conditions that cause pH gradient elution, wherein the The anion exchange medium comprises silica gel derivatized with cationic groups so that the prions are collected in an eluate fraction which is eluted from an anion exchange chromatography column at pH 6.5 to 8.3, thereby , so that the prions are separated from the collected proteins. Background technique [0002] Spongiform encephalopathies are disorders of the central nervous system in mammals that cause premature dementia and are generally fatal. Some of these encephalopathies involve humans, for example, cortico-striato-myeloid degeneration, kuru, parieto-occipital syndrome, sheep dysentery, bovine spongiform encephalopathy, etc. Although there is strong evidence that these ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/17C07K1/18C07K14/805
CPCC07K1/18C07K14/805
Inventor 玛丽亚·S·加瑞尔罗伯特·A·胡特陈斯威廉姆·R·赖特
Owner HEMOGLOBIN OXYGEN THERAPEUTICS
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