Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for directly extracting and purifying trasylol from ox lung

A technology of aprotinin and bovine lung, which is applied in the field of extracting and purifying aprotinin, which can solve the problems of unsuitability for industrial production, long production cycle and harsh use conditions

Inactive Publication Date: 2006-05-17
NANCHANG WANHUA BIOCHEM PHARMA
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When agarose beads are used as the matrix of affinity chromatography, the disadvantages are that the mechanical strength of the adsorbent is small, the use conditions are harsh, and the cost is high, so it is not suitable for industrial production.
The traditional chromatographic purification process has a long production cycle (each production batch takes more than three days, and the sample adsorption of affinity chromatography takes more than one day), and the product has low potency (only 5000- 6000KIU / mg), low recovery rate, many production steps, high labor intensity and other shortcomings
Moreover, although the traditional chromatography method has the advantages of high resolution, due to the limitation of the mechanical strength of the matrix, there are many shortcomings such as small column capacity, low flow rate, and low throughput, which are subject to certain restrictions in large-scale industrial production. limits

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0013] The following is a specific implementation example of the present invention.

[0014] Activation of methoxylated polyethylene glycol (MPEG): Weigh 100g of MPEG and 20g of anhydrous sodium carbonate, dissolve in 500mL of benzene, add 1.5g of s-trichlorotriazine, stir and reflux at 80°C for 24 hours, centrifuge, The supernatant was poured into petroleum ether (30-60° C.), and the activated MPEG2 crude product was precipitated. After suction filtration, the crude product was dissolved with a 1:1 volume ratio of benzene:acetone mixture, and then precipitated with petroleum ether. The same dissolution and precipitation were repeated 5 times, and the activated MPEG was obtained by draining.

[0015] Coupling of MPEG and trypsin after activation: Take 3.0 g of MPEG after activation and dissolve it in 0.025mol / L borax buffer, then add some trypsin (titer is more than 3000KIU / mg), stir gently (50rpm ), the coupling conditions are: 15° C., 8 hours, trypsin:PEG ratio of 1:5, pH ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

A method for directly extracting and purifying aprotinin from bovine lungs, which involves immobilizing trypsin on polyethylene glycol molecules and obtaining trypsin-PEG polymers with appropriate molecular weights, thereby establishing an affinity two-phase extraction system , the bovine lung is acid-hydrolyzed with sulfuric acid after removal of impurities, cut into pieces, mashed, and then filtered. After the filtrate is extracted with an affinity two-phase aqueous phase, the final upper phase is desalted by ultrafiltration and freeze-dried to obtain high-purity aprotinin product. The product prepared by the present invention meets the requirements of British Pharmacopoeia 98 edition for aprotinin products, that is, macromolecular substances cannot be detected (molecular sieve chromatography). The potency of the product reaches above 6300KIU / mg.

Description

technical field [0001] The invention relates to a method for directly extracting and purifying aprotinin from bovine lung, in particular to a method for directly extracting and purifying aprotinin from bovine lung by adopting an affinity two-phase extraction process. Background technique [0002] Aprotinin, also known as Bovine pancreatic trypsin inhibitor (BPTI), Kallikrein inhibitor, Kunitz inhibitor, and antiprotease peptide. Its molecular formula is C284H432N84079S7, which is a basic polypeptide chain containing 58 amino acid residues, its molecular weight is 6.5KD, and its isoelectric point is 10.5. Aprotinin is a white or slightly yellow powder, odorless, easily soluble in water, soluble in 70% ethanol and 50% acetone, etc., and has dialyzability. Aprotinin has been used since the 1960s, and it was found to have anti-kinin and anti-shock effects. Then, some people successfully used high-dose aprotinin to treat acute pancreatitis, which has ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00
Inventor 张万山刘兰花李菊根王启要
Owner NANCHANG WANHUA BIOCHEM PHARMA