Method for synthesizing terbutaline and bovine serum albumin conjugate

A technology for bovine serum albumin and terbutaline, which is applied in the field of synthesizing terbutaline and bovine serum albumin conjugates, can solve problems such as affecting health and poisoning, achieves short time, simple synthesis process, and simplifies synthesis effect of steps

Inactive Publication Date: 2017-07-04
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After people eat it, they will gradually be poisoned, produce abn

Method used

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  • Method for synthesizing terbutaline and bovine serum albumin conjugate
  • Method for synthesizing terbutaline and bovine serum albumin conjugate
  • Method for synthesizing terbutaline and bovine serum albumin conjugate

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Dissolve BSA (66.43 mg, 1 μmol) in PBS (pH=6.0) buffer, then add terbutaline (8.23 mg, 30 μmol), stir to dissolve, then add dropwise 40% formaldehyde solution (30 μl), at room temperature Stir the reaction for 24 hours to obtain the reaction solution; transfer the reaction solution to a dialysis bag, put the dialysis bag into a beaker filled with distilled water, and dialyze for 3 days under stirring at room temperature; centrifuge the dialysis product in a high-speed refrigerated centrifuge, and collect The clear liquid was lyophilized in vacuum for 12 hours to obtain a white powder of the conjugate (26.3 mg, yield: 39.42%).

[0022] BSA, terbutaline and the conjugate were prepared with PBS (pH=6.0) respectively to prepare 0.5 mg / ml solutions for later use. First use PBS buffer solution (pH=6.0) to balance the baseline in the wavelength range of 220-400nm, then scan the prepared 3 solutions respectively, and record the absorbance values ​​scanned at each wavelength ban...

Embodiment 2

[0024] After compressing BSA, terbutaline and the conjugated product with KBr, the infrared spectrum was scanned. Although the infrared absorption curve of the conjugated product is very similar to that of BSA, some absorption peaks have moved or The enhancement was presumed to be due to the coupling of BSA to terbutaline. From the infrared spectrum of terbutaline, it can be seen that the absorption of 1,3,5-trisubstituted aromatic hydrocarbons: 695cm -1 The strong absorption peak of the 1,3,5-trisubstituted compound at , the skeleton vibration of the benzene ring is at 1610cm -1 、1522cm -1 and 1486cm -1 These three absorption peaks can also be seen in -C(CH 3 ) 3 The methyl group of the structure has two absorption peaks with different intensities, namely about 1382cm -1 、1344cm -1 , 3334cm in the picture -1 The broad peak is the stretching vibration of the hydroxyl group, and the infrared spectra of the conjugates have similar absorption peaks. In addition, in the in...

Embodiment 3

[0026] image 3 Shown are the fluorescence effects of BSA, terbutaline, conjugates, and equimolar mixed solutions of terbutaline and BSA under 365nm ultraviolet light. The substances in the solutions of the first three groups are all 0.1 mM. The sum of BSA and terbutaline in the mixed solution was 0.1 mM. It can be clearly seen that only the conjugate has fluorescence, and it can also be inferred that the conjugation of terbutaline and BSA is successful. See image 3 , in the figure: (a) the fluorescence effect diagram of BSA, (b) the fluorescence effect diagram of terbutaline, (c) the fluorescence effect diagram of the conjugate, (d) the fluorescence effect of the mixture of terbutaline and BSA picture.

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Abstract

The invention provides a method for synthesizing terbutaline and bovine serum albumin conjugate. The method comprises the following steps: 1, dissolving bovine serum albumin (BSA) in a phosphoric acid buffering solution (PBS with the pH value of 6.0), adding terbutaline, stirring the terbutaline and the obtained solution to dissolve the terbutaline, dropwise adding a formaldehyde solution, and carrying out a stirring reaction at room temperature for 24 h to obtain a reaction solution; 2, transferring the reaction solution into a dialysis bag, placing the dialysis bag in a beaker provided with distilled water, and carrying out dialysis under a room temperature stirring condition for 3 days; and 3, centrifuging the above obtained dialysis product by a high speed refrigerated centrifuge, collecting the obtained supernatant, and carrying out vacuum freeze drying to obtain dry powder that is the terbutaline and bovine serum albumin conjugate. The terbutaline and bovine serum albumin conjugate is synthesized through one step, and is of great significance to the immunodetection exploitation and the application of a clenbuterol hydrochloride additive in foods.

Description

technical field [0001] The invention relates to a protein conjugate, in particular to a method for synthesizing a conjugate of terbutaline and bovine serum albumin. Background technique [0002] Clenbuterol is a feed additive, which can improve the conversion rate of feed and increase the lean meat rate of animals. It has a large dosage, slow metabolism, stable chemical properties, and will decompose above 150°C. After people eat it, they will gradually become poisoned, produce abnormal physiological reactions, and seriously affect their health. Terbutaline belongs to the catechol-type clenbuterol compounds. At present, 4-aminobenzidine is used as the linker, and terbutaline is coupled with BSA with 1,4-butanediol glycidyl ether and p-aminobenzoic acid to prepare terbutaline coupled with bovine serum albumin. things. This process requires at least two steps of synthesis, and also requires a variety of reagents such as 4-aminobenzidine, so it is necessary to develop a new ...

Claims

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Application Information

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IPC IPC(8): C07K14/765G01N33/531
CPCC07K14/765C07K19/00G01N33/5308G01N33/531
Inventor 杜会枝
Owner SHANXI UNIV
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