46 results about "Insulin glargine" patented technology

The invention relates to methods of separation and / or purification of impurities yielding a purified heterologous protein product devoid of related impurities or with substantially minimal quantities of such glycosylated impurities. More specifically, the invention relates to the identification of glycosylated forms of insulin analogues such as glargine impurities characterized post expression in yeast based systems such as Pichia pastoris. The invention also relates to methods used to clone gene encoding the protein insulin glargine; inserting the related gene in a suitable yeast host; producing culture of the recombinant strain, stimulating expression of the heterologous polypeptide, its secretion and purification post fermentation and related enzymatic conversions.

The invention relates to a method for preparing a recombinant glycine-arginine insulin crystal. The method specifically comprises the following steps of (1) dissolving recombinant glycine-arginine insulin, a phenol derivative, a zinc-containing substance and organic acid in water so as to obtain crystal fluid; and (2) taking a pH regulating agent for regulating the pH value of the crystal fluid to 4.5-7.0, performing stirring for 3-9 hours, and performing separation so as to obtain the recombinant glycine-arginine insulin crystal, wherein the concentration of the recombinant glycine-arginine insulin in the crystal fluid is 1-3.5g / L, the mass / volume percentage of the phenol derivative is 0.01-1.00%, the mass / volume percentage of the zinc-containing substance is 0.005-1.5%, and the mass / volume percentage of the organic acid is 0.01-2.0%. The crystal fluid prepared by the method does not contain organic solvents, the operation process is safe, the technology is simple and easy to control, poisonous and harmful substances in chromatography in the previous step can be effectively removed, the time for filtration, collection, crystal washing and freeze drying of products is shortened, the cost is reduced, and the method is suitable for industrialized production of the recombinant glycine-arginine insulin.

Disclosed is a method for preparing an insulin glargine (GlyA21-ArgB31-AryB32-human insulin) crystal, comprising crystallizing the insulin glargine at pH 7.0-9.0 and in a crystallization solution containing a recombinant insulin glargine, an organic solvent of a 10-30% concentration by volume, a zinc compound, a phenol derivative, a salt and an organic acid.

The invention relates to the field of biochemistry and discloses an extraction method of an insulin glargine precursor protein. The extraction method comprises the following steps of adding an acid into an insulin glargine precursor protein fermentation broth to adjust a pH value to 1-4, carrying out centrifugation and collecting a supernatant so that the insulin glargine precursor protein is obtained. Before bacterium centrifugation, the acid is used to adjust a pH value of the insulin glargine precursor protein fermentation broth to 1-4 so that after extraction, insulin glargine precursor protein content is improved, target protein loss caused by direct centrifugation removal of the bacteria is avoided and an insulin glargine yield is improved.

The invention provides a method for improving the efficiency in preparation of insulin and the like. The method comprises the step of conducting chromatography on a precursor of insulin glargine by use of a hydroxyapatite medium. The invention further provides a preparation method for active insulin glargine. The preparation method comprises the step of conducting enzyme digestion on a recombinant expression precursor of insulin glargine by use of a Kex-2p enzyme. In a preferred embodiment of the invention, the preparation method comprises the following steps: obtaining the recombinant expression precursor of insulin glargine; conducting enzyme digestion on the recombinant expression precursor of insulin glargine by use of the Kex-2p enzyme; conducting further chromatographic purification on the enzyme digestion product, so as to obtain active insulin glargine.

The invention relates to the technical field of insulin crystallization, in particular to a method for preparing insulin glargine crystals. The method for preparing insulin glargine crystals adopts the gas-phase diffusion hanging drop method, comprising the following steps: (1) preparing a crystallization solution containing the following components: Tris 0.2-1.0M, citric acid 0.2-1.0M; the pH of the crystallization solution is 8.0 to 11.0; (2) Mix the insulin glargine solution with the crystallization solution to obtain a mixed crystallization solution; (3) Hang the mixed crystallization solution above the mother liquor pool filled with the crystallization solution, and let it stand for cultivation . The method can be used to obtain insulin glargine crystals with regular shape and suitable size in the form of single crystals, and the resolution of X-ray diffraction is high, and the crystal structure of insulin glargine can be correctly analyzed.