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A method of purifying proteins

A protein, protein technology, applied in the field of protein purification

Inactive Publication Date: 2015-10-07
CSL BEHRING AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, while endogenous Hp and Hx control the side effects of free Hb / hemoglobin under physiological steady-state conditions, they maintain steady-state Hb / hemoglobin levels under pathophysiological conditions such as those associated with hemolysis. little effect

Method used

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Embodiment 1

[0134] Starting material: Cohn fraction IV was used 4 The precipitate was used as starting material for the purification of haptoglobin, hemopexin, and transferrin (see figure 1 ).

[0135] Precipitation extraction: by introducing 20 g extraction buffer / g Fraction IV 4 Precipitate (20x extraction rate), an extraction of the precipitate was performed. Mix the buffer and pellet for at least 1 h. Extraction buffer consisted of 50 mM Tris adjusted to pH 7.0 with concentrated HCl. Prepare the extraction buffer at a temperature of 20-25 °C. Precipitation extraction was also carried out at a temperature of 20-25°C. During the 1 hour extraction time, the pH during extraction was maintained at 7.0 to 8.0 (preferably 7.0).

[0136] Ammonium sulfate precipitation: to fraction IV 4 Solid ammonium sulfate is added to the extract in order to obtain a final concentration of 2.0 to 2.5M (preferably 2.4M). Ammonium sulfate was slowly added to the extract with stirring and mixing was co...

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Abstract

The present invention relates generally to a method of purifying proteins. More specifically, the present inventions relates to a method of purifying haptoglobin and hemopexin from the same starting material, and uses thereof.

Description

technical field [0001] The present invention generally relates to a method of purifying proteins. More particularly, the present invention relates to a method for purifying haptoglobin and hemopexin from the same starting material, and uses thereof. Background technique [0002] Hemolysis is characterized by the destruction of red blood cells and is a hall-mark of anemic conditions associated with red blood cell abnormalities, such as enzyme deficiencies, hemoglobinopathies, hereditary spherocytosis, paroxysmal nocturnal hemoglobinuria, and spinous erythrocytes (spur cell) anemia, and exogenous factors such as splenomegaly, autoimmune disorders (eg, hemolytic disease of the newborn), genetic disorders (eg, sickle cell disease or G6PD deficiency), microangiopathic hemolysis, genetic Lambert-positive bacterial infections (eg, streptococci, enterococci, and staphylococci), parasitic infections (eg, Plasmodium), toxins, and trauma (eg, burns). Hemolysis is also a common transf...

Claims

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Application Information

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IPC IPC(8): C07K14/79C07K1/14C07K1/16C07K1/30C07K1/36C07K14/47
CPCC07K1/14C07K1/16C07K1/30C07K1/36C07K14/79C07K14/4717C07K14/805A61P13/02A61P13/12A61P17/02A61P33/06A61P35/02A61P37/02A61P37/06A61P7/00A61P7/04A61P7/06Y02A50/30C07K1/18A61K38/1722A61K38/42
Inventor N·布林克曼
Owner CSL BEHRING AG
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