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Preparation method of S-adenosylhomocysteine-ovalbumin conjugate

A technology for homocysteine ​​and adenosine, which is applied in the field of preparation of S-adenosyl homocysteine-chicken ovalbumin conjugates, can solve the problem of complex process, small amount of coupling products and easy coupling agent. Overdose and other problems, to achieve the effect of simple process operation, high yield of conjugates, and good activity of conjugates

Inactive Publication Date: 2020-08-07
上海领潮生物新材料有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the preparation of conjugates by methods 1 and 2 is time-consuming, complicated and difficult to operate; although the process of method 3 is simple, glutaraldehyde is easy to form self-coupling between carriers, and the coupling agent is easy to be excessive, which is difficult to control Simultaneously, the above-mentioned method also has defects such as few coupling products, poor activity, and low utilization rate of the coupling agent.

Method used

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  • Preparation method of S-adenosylhomocysteine-ovalbumin conjugate
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Embodiment 1

[0029] This embodiment provides a preparation method of S-adenosylhomocysteine-chicken ovalbumin conjugate, the preparation method comprising the following steps:

[0030] (1) Activate chicken ovalbumin with suberic acid disuccinimide in dimethylformamide, the activation temperature is 37°C, and the activation time is 30min; wherein the chicken ovalbumin and the suberic acid di The molar ratio of succinimide is 1: 20;

[0031] (2) The mixture after the activation of step (1) is dialyzed successively with two dialysates, centrifuged at 5000rpm for 30s, and the precipitate is discarded to obtain the supernatant, and the concentration of the discarded supernatant is measured with an ultraviolet spectrophotometer; wherein the pH value is used first Dialyze for 7.4 phosphate buffer solution for 15 minutes, and then use sodium borate buffer solution with a pH value of 9.8 for 15 minutes;

[0032] (3) Add S-adenosylhomocysteine ​​to the supernatant obtained in step (2), add new disu...

Embodiment 2

[0036] This embodiment provides a preparation method of S-adenosylhomocysteine-chicken ovalbumin conjugate, the preparation method comprising the following steps:

[0037] (1) Activate chicken ovalbumin with suberic acid disuccinimide in dimethylformamide, the activation temperature is 37°C, and the activation time is 30min; wherein the chicken ovalbumin and the suberic acid di The molar ratio of succinimide is 1: 22;

[0038] (2) The mixture after the activation of step (1) is dialyzed successively with two dialysates, centrifuged at 5000rpm for 30s, and the precipitate is discarded to obtain the supernatant, and the concentration of the discarded supernatant is measured with an ultraviolet spectrophotometer; wherein the pH value is used first Dialyze for 7.4 phosphate buffer solution for 12 minutes, and then use sodium borate buffer solution with a pH value of 9.8 for 12 minutes;

[0039] (3) Add S-adenosylhomocysteine ​​to the supernatant obtained in step (2), add new disu...

Embodiment 3

[0043] This embodiment provides a preparation method of S-adenosylhomocysteine-chicken ovalbumin conjugate, the preparation method comprising the following steps:

[0044](1) Activate chicken ovalbumin with suberic acid disuccinimide in dimethylformamide, the activation temperature is 37°C, and the activation time is 35min; wherein the chicken ovalbumin and the suberic acid di The molar ratio of succinimide is 1: 18;

[0045] (2) The mixture after the activation of step (1) is dialyzed successively with two dialysates, centrifuged at 5000rpm for 30s, and the precipitate is discarded to obtain the supernatant, and the concentration of the discarded supernatant is measured with an ultraviolet spectrophotometer; wherein the pH value is used first Dialyze for 7.4 phosphate buffer solution for 10 minutes, and then use sodium borate buffer solution with a pH value of 9.8 for 10 minutes;

[0046] (3) Add S-adenosylhomocysteine ​​to the supernatant obtained in step (2), add new disuc...

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Abstract

The invention discloses a preparation method of an S-adenosylhomocysteine-ovalbumin conjugate. The preparation method comprises the following steps: (1) activating ovalbumin with succinimide octanedioate in a solvent; (2) sequentially dialyzing and centrifuging an activated mixture obtained in step (1) by adopting at least two dialyzates to obtain a supernatant; and (3) adding S-adenosylhomocysteine into the supernatant obtained in step (2), adding new succinimide octanedioate, mixing, and carrying out a coupling reaction to obtain the product. The S-adenosylhomocysteine-chicken ovalbumin conjugate can be rapidly and efficiently prepared with high quality (good activity), and the target conjugate is high in yield and good in activity.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, in particular to the application of S-adenosylhomocysteine ​​in immunological determination, and in particular to a preparation method of S-adenosylhomocysteine-chicken ovalbumin conjugate . Background technique [0002] Homocysteine ​​(Hcy) has always been regarded as an independent risk factor for cardiovascular disease. At present, enzyme-linked immunosorbent assay (ELISA) technology has been used in the determination of Hcy in the market, and the ELISA determination technology of Hcy has been established. One of the key steps is the preparation of S-adenosylhomocysteine ​​(SAH)-chicken ovalbumin (OVA) conjugate, S-adenosylhomocysteine ​​(SAH) is the precursor substance of Hcy, its It can be retrosynthesized by Hcy. Studies have also shown that perhaps SAH may be a more sensitive indicator of cardiovascular disease risk than Hcy. Increased SAH can induce damage to vascular endothelial ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/77C07K1/10
CPCC07K14/77
Inventor 罗朝领茅柳娟曹雪姣尹文偲倪伟青
Owner 上海领潮生物新材料有限公司
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