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SAM (S-Adenosyl-L-homocysteine) artificial complete antigen, preparation method and application

A cysteine ​​and adenosine isotype technology, applied in the field of organic chemistry and immunology, can solve the problems of inability to stabilize SAH antigen and antibody, lack of SAH molecular structure antigen preparation technology, etc., and achieve high specificity effect

Active Publication Date: 2020-08-14
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this key issue has been ignored in the few studies on the preparation of SAH and Hcy monoclonal antibodies. There is a lack of an optimized and reasonable antigen preparation process based on the molecular structure of SAH, and it is impossible to stably produce high-quality and efficient SAH antigens and antibodies.

Method used

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  • SAM (S-Adenosyl-L-homocysteine) artificial complete antigen, preparation method and application
  • SAM (S-Adenosyl-L-homocysteine) artificial complete antigen, preparation method and application
  • SAM (S-Adenosyl-L-homocysteine) artificial complete antigen, preparation method and application

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preparation example Construction

[0024] see figure 1 A preferred embodiment of the present invention provides a method for preparing an S-adenosylhomocysteine ​​artificial complete antigen, comprising the following steps:

[0025] Step S11, mixing S-adenosyl homocysteine ​​with di-tert-butyl dicarbonate to obtain a first intermediate, and said di-tert-butyl dicarbonate is used to protect S-adenosyl homocysteine Antigenic epitopes in acids.

[0026] Wherein, the epitope in S-adenosyl homocysteine ​​(SAH) is protected by di-tert-butyl dicarbonate (Boc). Such as figure 2 As shown, the SAH molecule is a combination of homocysteine ​​(Hcy) molecule and adenosine through the -S-C- bond, and the only carboxyl group in the molecule is located at the homocysteine ​​part, and together with its adjacent α-amino group, it forms a characteristic Characterizing said antigenic epitopes. During the subsequent preparation of the antigen, the α-amino group must not react. Therefore, the α-amino group of the cysteine ​​pa...

Embodiment 1

[0044] Step 1: Dissolve 8.5mg of sodium bicarbonate in 2mL of deionized water, add 19.25mg of SAH after complete dissolution, and then slowly add tetrahydrofuran:dioxane as an amphiphilic solubilizer at a ratio of 1:1 in an ice-bath environment After the dropwise addition, stir magnetically at 0°C for 15 minutes to observe the dissolution of SAH, then add 2mL of tetrahydrofuran solution dissolved with 11.5 μL di-tert-butyl dicarbonate and continue the reaction at 0°C for 40 minutes, then raise the temperature to 25 ℃ to continue the reaction in the dark for 18 hours. After the reaction, add 5 mL of ethyl acetate to extract three times, separate the water phase and freeze it at -20°C, and recover the crystals by freeze-drying with a lyophilizer. The product is the first intermediate (namely Boc-SAH). Save for later use.

[0045] In the second step, 10 mg of the first Boc-SAH intermediate prepared in the first step and 30 mg of BSA were dissolved in 3 mL of PBS solution with a ...

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Abstract

The present invention provides a preparation method of an SAM (S-Adenosyl-L-homocysteine) artificial complete antigen. The method comprises the following steps: SAM is mixed with di-tert-butyl dicarbonate to obtain a first Intermediate, wherein di-tert-butyl dicarbonate is used for protecting epitope in SAM; adding carrier protein to the first intermediate, and obtaining a second intermediate after the reaction; and removing di-tert-butyl dicarbonate in the second intermediate to obtain the SAM artificial complete antigen. The SAM artificial complete antigen prepared by the method has high specificity, selectivity and immunogenicity. The invention also provides the SAM artificial complete antigen prepared by the method and an application of the SAM artificial complete antigen to antibody preparation.

Description

technical field [0001] The invention relates to the technical fields of organic chemistry and immunology, in particular to an artificial complete antigen of S-adenosyl homocysteine, a preparation method and application thereof. Background technique [0002] In organisms, the activated methylate S-adenosyl-methionine (S-Adenosyl-L-methionine, SAM) is catalyzed by enzymes to transfer methyl groups to various proteins or nucleic acid molecules, thereby forming corresponding methyl groups The product of methylation is called biological methylation. The demethylated SAM generates S-adenosyl-homocysteine ​​(S-Adenosyl-L-homocysteine, SAH), and then undergoes further hydrolysis to remove adenosine to generate homocysteine ​​(homocysteine, Hcy). It is the only way to produce Hcy in vertebrates known so far. [0003] Methylation is an important biochemical reaction ubiquitous in organisms, closely related to gene regulation, body metabolism, aging, disease, hormones and other life ...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K14/77C07K14/795C07K16/44C07K1/06
CPCC07K14/765C07K14/77C07K14/795C07K16/44Y02P20/55
Inventor 马岚刘丹吴峰岑瑜
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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