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Synthesis method of half-antigen and complete antigen of tulathromycin

A telamycin and complete antigen technology, applied in the field of immunology, can solve the problems of no telamycin enzyme-linked immunosorbent assay method, etc., and achieve the effect of high titer, simple preparation method and good specificity

Active Publication Date: 2015-06-17
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no relevant reports on telamycin enzyme-linked immunosorbent assay (ELISA) at home and abroad.

Method used

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  • Synthesis method of half-antigen and complete antigen of tulathromycin
  • Synthesis method of half-antigen and complete antigen of tulathromycin
  • Synthesis method of half-antigen and complete antigen of tulathromycin

Examples

Experimental program
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Effect test

Embodiment 1

[0025] The synthesis of embodiment 1 telamycin hapten

[0026] Use the succinic anhydride method to derivatize telamycin, and the specific steps are as follows: take 4 mg telamycin standard and dissolve it in 500 μl anhydrous pyridine, add 1 mg succinic anhydride, reflux and stir for 6 hours at 114 ° C, and dry the reaction solution with nitrogen , That is, the hapten of Tyramycin.

[0027] The obtained telamycin hapten was identified by LC-MS. The mass spectrum of the telamycin hapten is shown in figure 2 shown. The molecular weight of telamycin is 806.08, and the molecular weight of succinic anhydride is 100.07. According to theoretical speculation, the molecular weight of the derivative obtained after the reaction of the hydroxyl group in the telamycin structure with succinic anhydride is 906.08. From figure 2 It can be seen that there is a peak with a mass-to-charge ratio of 906.6 in the derivative, which is consistent with the peak obtained in theory and practice, ...

Embodiment 2

[0028] Synthesis of embodiment 2 telamycin hapten

[0029] Dissolve 5 mg of standard telamycin in 400 μL of anhydrous pyridine, add 2 mg of succinic anhydride, reflux and stir at 90-114°C for 9 hours, and dry the reaction solution to obtain the telamycin hapten.

Embodiment 3

[0030] The synthesis of the complete antigen of embodiment 3 Tyramycin

[0031] The complete antigen was synthesized by the active ester method. 1 mg of the product derived from telamycin (ie, hapten), 0.6 mg of EDC, and 0.4 mg of NHS were dissolved in 300 μl of DMF, and stirred at room temperature for 30 min in the dark. 6 mg of BSA was dissolved in 2 mL of PBS, and the activated telamycin derivative was added dropwise to the BSA solution while stirring, and reacted overnight at 4°C. After the reaction, dialyze with 0.01mol / L PBS for 3 days and change the solution every 6 hours.

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Abstract

The invention discloses a synthesis method of a half-antigen and a complete antigen of tulathromycin and belongs to the technical field of immunology. The method synthesizes the half-antigen of tulathromycin by carrying out hydroxyl on tulathromycin by a succinic anhydride method and introducing free carboxyl. Derivated free carboxyl reacts with primaquine of bovine serum albumin to form an amido bond for coupling by an active ester method to form the complete antigen of tulathromycin. After the antigens are used for immunizing a Balb / c mouse for three times, the valence can reach 1:8000. The antigens have wide practical application prospects.

Description

technical field [0001] The invention relates to a method for synthesizing hapten and complete antigen of telamycin, which belongs to the technical field of immunology. Background technique [0002] Tulathromycin (Tulathromycin, C 41 h 79 N 3 o 12 ), a semi-synthetic macrolide antibiotic approved for use in China in 2008. Tyramycin has strong drug efficacy and low dosage, and will gradually replace existing macrolide antibiotics such as tylosin and tilmicosin, which have developed drug resistance, and has great application potential. [0003] The research on the analysis method of telamycin residue is very necessary. At present, radiolabeled assays and HPLC-MS-MS are mainly used for the determination of telamycin residues. The latter is more commonly used, but the sample pre-treatment is complicated, the measurement process is cumbersome, and the cost of the instrument is high, which cannot meet the requirements of on-site quick inspection. The immunoassay method has t...

Claims

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Application Information

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IPC IPC(8): C07H17/00C07H1/00C07K14/765C07K14/77C07K16/44G01N33/68
CPCC07H1/00C07H17/00C07K14/765C07K14/77C07K16/44G01N33/6857
Inventor 孙秀兰朱欣凤张银志
Owner JIANGNAN UNIV
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