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A kind of hepatitis B virus adsorbent and its preparation method and application

A hepatitis B virus and adsorbent technology, applied in the field of biomedical adsorption materials, can solve the problems of long time, high immunogenicity of antibodies, high cost of antibodies, etc., and achieve the effects of long storage period, high adsorption efficiency and specific affinity

Active Publication Date: 2022-05-17
WUHAN RUIFA MEDICAL DEVICES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in this patent, antibodies are used as ligands, and the production of antibodies requires high costs and a long time. Antibodies will undergo irreversible denaturation after a certain period of time at room temperature or higher temperatures, and antibodies have high immunogenicity of

Method used

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  • A kind of hepatitis B virus adsorbent and its preparation method and application
  • A kind of hepatitis B virus adsorbent and its preparation method and application
  • A kind of hepatitis B virus adsorbent and its preparation method and application

Examples

Experimental program
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preparation example Construction

[0029] The present invention provides a method for preparing a hepatitis B virus adsorbent, comprising the following steps:

[0030] S1. Clean the carboxylated agarose gel microspheres to obtain an agarose gel microsphere suspension;

[0031] S2, the amino-modified nucleic acid aptamer is placed in the aptamer dissolution buffer, shaken and dissolved after water bath treatment, and then placed in a room temperature environment, after its reconstitution to obtain the dissolved aptamer;

[0032] S3, N-hydroxysuccinimide and 1- (3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride according to a preset molar ratio mixed in 2- (N- morpholine) ethanesulfonic acid buffer solution to obtain a crosslinking catalyst;

[0033] S4, the crosslinking catalyst obtained in step S3 was added to the agarose gel suspension obtained in step S1, mixed well and then added the dissolved aptamer obtained in step S2, for amide reaction;

[0034] S5, after the completion of the amide reaction in step ...

Embodiment 1

[0043] The present embodiment provides a method for preparing a hepatitis B virus adsorbent, the reaction principle thereof, as in Figure 1 As shown, the specific steps include the following steps:

[0044] S1, the carboxylated agarose gel microspheres shock mixed, using a pipette to extract 10 mL of mixed microspheres, transferred to a 100 mL Erlenmeyer flask, let stand at room temperature for 5 min, slowly aspirate and discard the upper protective buffer, add 30 mL distilled water to resuspend, transfer to the filter bottle, rinse three times with 200 mL distilled water to completely remove part of the protective buffer, and quickly transfer the washed microspheres to the 100 mL Erlenmeyer flask, the whole process keeps the microspheres moist, Obtain an agarose gel microsphere suspension.

[0045]S2, from the refrigerator of -20 °C, take out the EP tube containing 250 μg of amino modified nucleic acid aptamer, let it stand at room temperature for 5 min, centrifuge at 5000 r / mi...

Embodiment 2～10

[0050] Examples 2 to 10 provide a method for preparing hepatitis B virus adsorbents, compared with Example 1, the difference is that the molar ratio of NHS and EDC in step S3 or the reaction time in step S4 is changed, and the molar ratio and reaction time corresponding to each embodiment are shown in Table 1.

[0051] Table 1 Example 2 to 10 corresponding to the reaction parameters

[0052]

[0053]

[0054]Take a static blood sample from a hepatitis B patient, respectively, using the hepatitis B virus adsorbent prepared in Example 1 to 10, adsorb the blood sample according to the ratio of adsorbent: blood = 1:5, and the content of hepatitis B surface antigen in the sample before and after the adsorption is detected. Wherein, the content of hepatitis B surface antigen in the sample before adsorption is 66.29 IU / mL, respectively, the content of hepatitis B surface antigen obtained after adsorption using the hepatitis B virus adsorbent provided in Example 1 to 10 is shown in ...

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Abstract

The invention provides a hepatitis B virus adsorbent as well as its preparation method and application. Crosslinking catalyst is prepared by mixing N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride in a buffer solution; and Add the cross-linking catalyst of the carboxylated agarose gel microsphere suspension into the carboxylated agarose gel microsphere suspension, mix well and then add the dissolved amino-modified nucleic acid aptamer, and use the amidation reaction to combine the carboxylated agarose gel microsphere with the amino group The modified nucleic acid aptamer is directly coupled, so that the hepatitis B virus adsorbent is prepared simply and efficiently. Through the above method, the present invention can use nucleic acid aptamers instead of traditional antibodies as ligands, so that the prepared adsorbent has higher adsorption efficiency, and realizes effective adsorption of hepatitis B surface antigen, thereby effectively reducing the amount of hepatitis B in blood. The content of hepatitis surface antigen has high practical application value.

Description

Technical field [0001] The present invention relates to the field of biomedical adsorption material technology, in particular to a hepatitis B virus adsorbent and preparation method and application thereof. Background [0002] Hepatitis B is a liver disease, both acute and chronic, caused by hepatitis B virus (HBV) infection. Positive seros test for HBV surface antigen (HBsAg) is the main criterion for confirming the diagnosis, and in clinical practice, clearance of HBsAg is considered to be the most critical therapeutic endpoint for chronic hepatitis B, which is associated with improving clinical outcomes, prolonging survival, and reducing the incidence of cirrhosis and hepatocellular carcinoma. At present, treatments for hepatitis B are mainly focused on drug screening, administration and targeted therapy, but these treatment options have limited effect and long-term treatment is not effective. [0003] Based on the problems existing in existing treatment options, in vitro blo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/30B01J20/24B01J20/28A61M1/36
CPCB01J20/3071B01J20/3085B01J20/24B01J20/28021A61M1/3679A61M1/362B01J2220/4856
Inventor 王业富张磊郑豪
Owner WUHAN RUIFA MEDICAL DEVICES CO LTD
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