Method of purifying apolipoprotein A-1
OwnerKieu Hoang, Bao Xiangfeiinventors
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Abstract
A first method of purifying apolipoprotein A-1 includes mixing plasma fraction IV with a 1-8 M urea solution to form a pretreatment solution; loading the pretreatment solution to a first anion chromatography column, and then eluting to obtain an apoA-1 protein solution; and loading the apoA-1 protein solution to a second anion chromatography column, and eluting to obtain pure apoA-1 protein. A second method includes dissolving plasma fraction IV in a buffer to produce a pretreatment solution; adding NaCl to the pretreatment solution and cooling it to form apoA-1 precipitate; collecting and reconstituting the apoA-1 precipitate; loading the reconstituted apoA-1 to an anion exchange column; and eluting apoA-1 from the column.
Claims33
A method of purifying apolipoprotein A-1, comprising:
- a) mixing plasma fraction IV with a solution of 1-8 M urea, forming a fraction IV pretreatment solution;
- b) loading the pretreatment solution acquired in step a) to a first anion exchange chromatography column, and then eluting with a buffer I comprising 1-8 M urea having a conductivity of 1-4 mS/cm to obtain an eluate I that mainly contains apoA-1 protein;
- c) loading eluate I to a second anion exchange chromatography column, first eluting with a buffer II, having a conductivity of 1-15 mS/cm, and later eluting with a buffer III, having a conductivity of 30-100 mS/cm, to obtain pure apoA-1 protein.
A method of purifying apolipoprotein A-I, comprising:
- a) dissolving plasma fraction IV in a buffer to produce a fraction IV pretreatment solution;
- b) centrifuging the pretreatment solution to remove celite;
- c) adding NaCl to the pretreatment solution and cooling the pretreatment solution to form apoA-1 precipitate;
- d) again centrifuging the pretreatment solution, to collect the apoA-I precipitate;
- e) reconstituting the apoA-I precipitate;
- f) filtering nonsoluble substances from the reconstituted apoA-1;
- g) loading the reconstituted apoA-1 to an anion exchange chromatography column to combine protein in the reconstituted apoA-I, including apoA-I, onto the column;
- and
- h) eluting apoA-I from the column with a buffer.
Dependencies are inferred from the claim text.