Hepatitis B virus adsorbent as well as preparation method and application thereof

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

Active Publication Date: 2021-04-30
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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  • Hepatitis B virus adsorbent as well as preparation method and application thereof
  • Hepatitis B virus adsorbent as well as preparation method and application thereof
  • Hepatitis B virus adsorbent as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0029] The invention provides a kind of preparation method of hepatitis B virus adsorbent, comprises the steps:

[0030] S1, washing the carboxylated agarose gel microspheres to obtain an agarose gel microsphere suspension;

[0031] S2, placing the amino-modified nucleic acid aptamer in the aptamer dissolving buffer, shaking and dissolving, performing a water bath treatment, and then placing it in a room temperature environment, and obtaining the dissolved aptamer after renaturation;

[0032] S3, N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride are mixed according to the preset molar ratio and dissolved in 2-(N- Morpholine) in ethanesulfonic acid buffer solution, obtain cross-linking catalyst;

[0033] S4. Add the cross-linking catalyst obtained in step S3 into the agarose gel suspension obtained in step S1, mix well, then add the dissolved aptamer obtained in step S2, and perform amide reaction;

[0034] S5. After the amide reaction in st...

Embodiment 1

[0043] This embodiment provides a kind of preparation method of hepatitis B virus adsorbent, its reaction principle is as follows figure 1 As shown, it specifically includes the following steps:

[0044] S1. Shake and mix the carboxylated agarose gel microspheres, use a pipette gun to extract 10mL of the mixed microspheres, transfer them to a 100mL Erlenmeyer flask, let stand at room temperature for 5min, slowly absorb and discard the upper layer Protect buffer, add 30mL distilled water to resuspend, transfer to a suction filter flask, use 200mL distilled water to wash three times to completely remove part of the protection buffer, and quickly transfer the washed microspheres to a 100mL Erlenmeyer flask, the whole process makes The microspheres were kept wet, resulting in an agarose gel microsphere suspension.

[0045] S2. Take out the EP tube containing 250 μg of amino-modified nucleic acid aptamers from the refrigerator at -20°C, let it stand at room temperature for 5 minut...

Embodiment 2～10

[0050] Embodiments 2 to 10 respectively provide a preparation method of a hepatitis B virus adsorbent. 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 are changed, The molar ratios and reaction times corresponding to each embodiment are shown in Table 1.

[0051] The reaction parameters corresponding to Table 1 Embodiment 2~10

[0052]

[0053]

[0054] Get the static blood sample from the hepatitis B patient, respectively adopt the hepatitis B virus adsorbent prepared in embodiment 1~10, according to the ratio of adsorbent:blood=1:5, the blood sample is adsorbed, and the hepatitis B surface in the sample before and after adsorption Antigen content was detected. Among them, the content of hepatitis B surface antigen in the sample before adsorption was 66.29IU / mL, and the content of hepatitis B surface antigen obtained after adsorption with the hepatitis B virus adsorbents provided in Examples 1-10...

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Abstract

The invention provides a hepatitis B virus adsorbent as well as a preparation method and application thereof. The preparation method comprises the following steps: mixing Nhydroxysuccinimide and 1 (3-dimethylaminopropyl) 3ethylcarbodiimide hydrochloride, and dissolving the mixture in a buffer solution to prepare a cross-linking catalyst; adding the obtained cross-linking catalyst into a carboxylated agarose gel microsphere suspension, fully and uniformly mixing, then adding a dissolved amino-modified nucleic acid aptamer, and directly coupling the carboxylated agarose gel microsphere with the amino-modified nucleic acid aptamer by virtue of amidation reaction, and therefore, the hepatitis B virus adsorbent is simply, conveniently and efficiently prepared. By means of the mode, the nucleic acid aptamer can replace a traditional antibody to serve as a ligand, the prepared adsorbent has higher adsorption efficiency, effective adsorption of the hepatitis B surface antigen is achieved, and therefore the content of the hepatitis B surface antigen in blood is effectively reduced, and high practical application value is achieved.

Description

technical field [0001] The invention relates to the technical field of biomedical adsorption materials, in particular to a hepatitis B virus adsorbent and its preparation method and application. Background technique [0002] Hepatitis B is a liver disease caused by hepatitis B virus (HBV) infection, including acute and chronic hepatitis. HBV surface antigen (HBsAg) seropositive is the main criterion for diagnosis. In clinical practice, clearing HBsAg is considered to be the most critical treatment endpoint of chronic hepatitis B, which is associated with improved clinical outcomes, prolonged survival, and reduced liver cirrhosis and hepatocytes. associated with the incidence of cancer. At present, the treatment methods for hepatitis B mainly focus on drug screening, drug administration and targeted therapy, etc., but the effects of these treatment options are limited, and the long-term treatment effect is not good. [0003] Based on the problems existing in the existing tr...

Claims

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Application Information

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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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