Coupling compound of tetracycline, preparation process and application thereof

A technology of tetracycline and conjugates, applied in chemical instruments and methods, biological testing, animal/human peptides, etc., can solve problems such as high price, short shelf life, and inability to meet detection needs, and achieve long-term compensation and saving The effect of testing time

Inactive Publication Date: 2006-11-15
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After searching, there are no reports on the synthesis of tetracycline immunogens and antibody preparation in the world, and most of the domestic ELISA kits for detecting tetracycline drug residues are pu

Method used

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  • Coupling compound of tetracycline, preparation process and application thereof
  • Coupling compound of tetracycline, preparation process and application thereof
  • Coupling compound of tetracycline, preparation process and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0041] (1) Preparation of solution A: Dissolve 40 mg of tetracycline in 8 ml of anhydrous acetone, pass through nitrogen, then quickly add 29 mg of 1,1'-carbonyldiimidazole, and react in the dark at 37 ° C for 3 hours under nitrogen protection. Reaction generates the active intermediate of tetracycline and 1,1'-carbonyldiimidazole (CDI), for subsequent use; meanwhile, take 200mg bovine serum albumin (molecular weight is 6.8Kda) for subsequent use;

[0042](2) Preparation of solution B: the acetone in the above-mentioned reacted solution A was spun to dryness, and 15ml of pH=8.5 was added, and the concentration was 0.5M borax buffer solution to dissolve the generated tetracycline and the activity of 1,1'-carbonyldiimidazole intermediate, and then add the above-mentioned bovine serum albumin, so that the final concentration of bovine serum albumin is 13mg / ml, under the protection of nitrogen, 30 ℃ dark reaction for 48 hours; Stir dialysate solution B at -4°C for 72 hours, then d...

Embodiment 2

[0045] (1) Preparation of solution A: Dissolve 40 mg of tetracycline in 8 ml of anhydrous acetone, pass through nitrogen, then quickly add 16 mg of 1,1'-carbonyldiimidazole, and react in the dark at 37°C for 2 hours under nitrogen protection. Reaction generates the active intermediate of tetracycline and 1,1'-carbonyldiimidazole (CDI), for subsequent use; meanwhile, take 240mg bovine serum albumin (molecular weight is 6.7Kda) for subsequent use;

[0046] (2) Preparation of solution B: the acetone in the above-mentioned reacted solution A was spun to dryness, and 20ml of pH=9.0 was added, and the concentration was 0.5M borax buffer solution to dissolve the generated tetracycline and 1,1'-carbonyldiimidazole activity Intermediate, and then add the above-mentioned bovine serum albumin, so that the final concentration of bovine serum albumin is 12 mg / ml, under the protection of nitrogen, 25 ℃ dark reaction for 36 hours; Stir dialysate solution B at -4°C for 70 hours, then dialyze ...

Embodiment 3

[0049] (1) Preparation of solution A: Weigh 20 mg of tetracycline and dissolve it in 5 ml of anhydrous acetone, pass through nitrogen, then quickly add 10 mg of 1,1'-carbonyldiimidazole, and react in the dark at 37°C for 4 hours under the protection of nitrogen, Reaction generates the active intermediate of tetracycline and 1,1'-carbonyldiimidazole (CDI), for subsequent use; meanwhile, take 155mg bovine serum albumin (molecular weight is 6.9Kda) for subsequent use;

[0050] (2) Preparation of Solution B: Rotate the acetone in the A solution after the above reaction to dryness, add 15ml pH=8.0, and the concentration is 0.5M borax buffer solution to dissolve the generated tetracycline and 1,1'-carbonyldiimidazole activity intermediate, and then add the above-mentioned bovine serum albumin, so that the final concentration of bovine serum albumin is 10mg / ml, under the protection of nitrogen, 20 ℃ dark reaction for 40 hours; Stir dialysate solution B at -4°C for 75 hours, then dial...

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Abstract

A conjugate of tetracycline is prepared from tetracycline semi-antigen and the bovine serum albumin (or egg albumin) as the carrier able to generate immunogenicity through coupling. It can be used for preparing the reagent kit for the enzyme-linked immunoassay of tetracycline.

Description

technical field [0001] The invention relates to a conjugate of antibiotics and a preparation method and application thereof, in particular to a tetracycline conjugate and a preparation method and application thereof. The invention belongs to the field of antibiotic immunoassay. Background technique [0002] The following titles involved in the present invention apply to the entire specification and claims: [0003] Bovine Serum Albumin (BSA): Product of Sigma [0004] Ovalbumin (OVA for short): a product of Sigma [0005] Phosphate Buffered Saline (PBS for short) (0.01M, pH=7.40) [0006] Dialysis membrane: Bioshorp products [0007] 1,1'-carbonyldiimidazole (1,1'-carbonyldiimidazole, referred to as CDI): Fluka company product [0008] Tetracycline: product of Sigma [0009] Hydroxysuccinimide (NHS for short): a product of Sigma [0010] Ethyl[3-(dimethylamino)propyl]carbodiimide (EDC for short): product of Sigma [0011] p-aminobenzoic acid (4-aminobenzoic acid): pr...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07C237/26C07K16/00G01N33/53
Inventor 郗日沫张玉兰卢圣欣
Owner SHANDONG UNIV
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