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Method for purifying chondroitin sulfate by utilizing immunoaffinity technology

A technology of chondroitin sulfate and immunophile, which is applied in the field of biomedicine, can solve the problems of low purity of chondroitin sulfate, and achieve the effects of easy promotion, strong retention and concentration, and short production cycle

Inactive Publication Date: 2015-03-04
JINAN SHENGYUELIN BIOLOGICAL ENG
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  • Claims
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Problems solved by technology

[0006] The invention provides a method for purifying chondroitin sulfate using immunoaffinity technology to solve the problem of low purity of chondroitin sulfate purified by alcohol precipitation method in the prior art

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  • Method for purifying chondroitin sulfate by utilizing immunoaffinity technology
  • Method for purifying chondroitin sulfate by utilizing immunoaffinity technology
  • Method for purifying chondroitin sulfate by utilizing immunoaffinity technology

Examples

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Embodiment 1

[0043] a. mice were subcutaneously injected with the chondroitin sulfate purified by alcohol precipitation method obtained in the comparative example, so that the mice produced an immune response; specifically, 180 micrograms of chondroitin sulfate purified by alcohol precipitation method were weighed and dissolved in 500 microliters of physiological In saline, mixed with an equal volume of Freund's complete adjuvant, fully emulsified, and subcutaneously immunized 3 Balb / c mice for the first time; every 2 weeks, weighed 180 micrograms of chondroitin sulfate purified by alcohol precipitation, Dissolved in 500 microliters of normal saline, and then emulsified with an equal volume of incomplete Freund's adjuvant for the second and third immunizations; before the fourth booster immunization, blood was collected from the tail of the mice, and the serum was separated. Coat the microtiter plate with chondroitin sulfate, measure the anti-chondroitin sulfate polyclonal antibody titer in...

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Abstract

The invention discloses a method for purifying chondroitin sulfate by utilizing immunoaffinity technology, belongs to the technical field of biological medicines, and aims at solving the problem that an alcohol method in the prior art is low in purity of purified chondroitin sulfate. The method comprises the following steps: 1) taking chondroitin sulfate as an immunogen to immunize a mouse, so as to prepare an anti-chondroitin sulfate monoclonal antibody; 2) coupling the anti-chondroitin sulfate monoclonal antibody with a solid-phase carrier, and taking the conjugate as an immunoaffinity column filling material to prepare a chondroitin sulfate immunoaffinity column; and 3) purifying chondroitin sulfate through the chondroitin sulfate immunoaffinity column. The screened-out antibody used for capturing chondroitin sulfate possesses high specificity, has extremely strong delaying and concentrating capacity on chondroitin sulfate in a sample, a chondroitin sulfate sample subjected to separation purification by using an affinity chromatography media coupled with the antibody has the advantages of high purity, good quality, environment friendliness, energy saving, short production period and the like, and the method is suitable for large-scale production and easy to popularize.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for purifying chondroitin sulfate by using immunoaffinity technology. Background technique [0002] Chondroitin sulfate is a class of glycosaminoglycans covalently linked to proteins to form proteoglycans, which are present in all organisms except plants. The related products developed with it as the main raw material are mainly used in the pharmaceutical industry. Therefore, there are relatively strict requirements for the purity of chondroitin sulfate and the limits of impurities therein. [0003] At present, chondroitin sulfate is mainly extracted from the cartilage of animals (such as sharks, cattle, chickens, etc.), the extraction method is mainly enzymatic hydrolysis, and the impurity removal method is mainly alcohol precipitation method. The chondroitin sulfate produced by the above process generally has problems such as low purity, cumbersome process, long p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08
Inventor 张敉张金阳于美妍李宝才姜福勤刘珊珊赵忠莹
Owner JINAN SHENGYUELIN BIOLOGICAL ENG
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