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Vitamin B5 conjugate and preparation method and application thereof

A technology of conjugates and vitamins, applied in the field of vitamin B5 conjugates and its preparation

Inactive Publication Date: 2015-11-25
TIANJIN SUNGENE BIOTECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching, there is no VB in China at present 5 ELISA kit for content detection, so research VB 5 Immunogen synthesis and antibody preparation are very necessary

Method used

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  • Vitamin B5 conjugate and preparation method and application thereof
  • Vitamin B5 conjugate and preparation method and application thereof
  • Vitamin B5 conjugate and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) Preparation of Solution A: In an ice bath, weigh 10 mg vitamin B 5 Dissolve in 1.3ml N,N-dimethylformamide (DMF), add tri-n-butylamine dropwise to it after completely dissolved, stir for 0.5h, then add 3.82μl isobutyl chloroformate dropwise, react for 1h, and react to form active Intermediate;

[0038] (2) Preparation of solution B: according to KLH and vitamin B 5 Weigh KLH with a mass ratio of 1:1 and dissolve it in 2ml of phosphate buffer solution with a pH of 7.38~7.56 and a concentration of 0.01M~0.02M to make a 5mg / ml solution for later use;

[0039](3) Take solution A and solution B respectively, add solution A dropwise to solution B under constant stirring at a temperature of 4°C, and react overnight to obtain solution C;

[0040] (4) Solution C was dialyzed with the above phosphate buffer solution for 70-80 hours with stirring, and then dialyzed with distilled water for 20-30 hours, and the dialysate was replaced every 6 hours;

[0041] (5) Freeze-dry th...

Embodiment 2

[0043] (1) Preparation of Solution A: In an ice bath, weigh 10 mg vitamin B 5 Dissolve in 1.3ml N,N-dimethylformamide (DMF), add tri-n-butylamine dropwise to it after completely dissolved, stir for 0.5h, then add 3.82μl isobutyl chloroformate dropwise, react for 1h, and the reaction generates active Intermediate;

[0044] (2) Preparation of solution B: according to KLH and vitamin B 5 Weigh KLH with a mass ratio of 2:1 and dissolve it in 2ml of phosphate buffer solution with a pH of 7.38~7.56 and a concentration of 0.01M~0.02M to make a 5mg / ml solution for later use;

[0045] (3) Take solution A and solution B respectively, add solution A dropwise to solution B under constant stirring at a temperature of 4°C, and react overnight to obtain solution C;

[0046] (4) Solution C was dialyzed with the above phosphate buffer solution for 70-80 hours with stirring, and then dialyzed with distilled water for 20-30 hours, and the dialysate was replaced every 6 hours;

[0047] (5) Fre...

Embodiment 3

[0049] (1) Preparation of Solution A: In an ice bath, weigh 10 mg vitamin B 5 Dissolve in 1.3ml N,N-dimethylformamide (DMF), add tri-n-butylamine dropwise to it after completely dissolved, stir for 0.5h, then add 3.82μl isobutyl chloroformate dropwise, react for 1h, and the reaction generates active Intermediate;

[0050] (2) Preparation of solution B: according to KLH and vitamin B 5 Weigh KLH with a mass ratio of 3:1 and dissolve it in 2ml of phosphate buffer solution with a pH of 7.38~7.56 and a concentration of 0.01M~0.02M to prepare a 5mg / ml solution for later use;

[0051] (3) Take solution A and solution B respectively, add solution A dropwise to solution B under constant stirring at a temperature of 4°C, and react overnight to obtain solution C;

[0052] (4) Solution C was dialyzed with the above phosphate buffer solution for 70-80 hours with stirring, and then dialyzed with distilled water for 20-30 hours, and the dialysate was replaced every 6 hours;

[0053] (5) ...

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Abstract

The invention discloses a vitamin B5 (VB5, (R)-N-(3,3-dimethyl-2,4-dyhydroxy-1-oxobutyl)-3-calcium alanine) conjugate. The vitamin B5 conjugate is formed by coupling vitamin B5 hapten and keyhole limpet hemocyanin (KLH) which generates immunogenicity and is a carrier material, wherein n refers to number of molecules, of VB5, combined with one keyhole limpet hemocyanin molecule and is an integer, KLH refers to keyhole limpet hemocyanin, and molecular weight ranges from 450KDa to 1300KDa. The invention further discloses a preparation method of the vitamin B5 conjugate. The preparation method includes: connecting vitamin B5 with the carrier material generating immunogenicity to form the coupling material capable of inducing an animal immune system to generate antibodies. The preparation method has the advantages of convenience, quickness, specificity and accuracy. A wide application space is provided for using the coupling material to manufacture an enzyme-linked immunosorbent assay reagent box of VB5.

Description

technical field [0001] The invention relates to a vitamin conjugate and its preparation method and application, in particular to vitamin B 5 Conjugates and their preparation methods and applications. It is a vitamin immune detection system. Background technique [0002] The following titles involved in the present invention apply to the entire specification and claims: [0003] KLH: KeyholeLimpetHemocyanin, Sigma [0004] PBS: PhosphateBafferedSaline (0.01M, pH=7.40) [0005] Vitamin B 5 : Tianjin Heowns company product [0006] Tri-n-butylamine: product of Tianjin Guangfu Fine Chemical Research Institute [0007] Isobutyl chloroformate: Beijing Bailingwei (J&K) company product [0008] Dialysis bag: product of Beijing Solarbio Co., Ltd. [0009] VB 5 It is a kind of vitamin B group, also known as "pantothenic acid" or "pantothenic acid". The main function is to participate in the production of energy in the body, which can control the metabolism of fat. It is als...

Claims

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Application Information

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IPC IPC(8): C07K14/795C07K1/107C07K16/44
CPCC07K14/795C07K16/44C07K19/00
Inventor 郗日沫许坤孟萌尹永梅张丽沙李小刚董亚庆王玉芬宋兆瑞龙浩
Owner TIANJIN SUNGENE BIOTECH
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