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Synthesis method of abietic acid artificial antigen

A technology of artificial antigen and synthesis method, applied in the field of synthesis of rosin acid artificial antigen, can solve the problems such as unsatisfactory titer of polyclonal antibody, and achieve the effect of concise synthesis steps

Inactive Publication Date: 2018-05-29
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

In the early stage, the inventor used 12-HA as the hapten, KLH and BSA as the carrier protein, and synthesized the complete antigen by the succinic anhydride method, but the titer of the polyclonal antibody obtained by immunizing rabbits was not ideal

Method used

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  • Synthesis method of abietic acid artificial antigen
  • Synthesis method of abietic acid artificial antigen
  • Synthesis method of abietic acid artificial antigen

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Embodiment Construction

[0033] 1. Adipate monoacylation of carrier protein

[0034] (1) Adipic monoacylation of KLH

[0035] According to the mass ratio of KLH:AA:DEC=1:10:3, weigh 50mg KLH, 500mgAA and 150mgDEC respectively. KLH was dissolved in anhydrous dimethylformamide (DMF) in advance, AA and DEC were added slowly under stirring at room temperature, and stirred overnight at room temperature. The reaction solution was transferred to a dialysis bag and dialyzed in distilled water for 48 hours, during which distilled water (4×2L) was replaced. The dialyzed solution was lyophilized to afford AA-KLH (47.6 mg).

[0036] (2) Succinic Acylation of BSA

[0037]According to the mass ratio of BSA:AA:DEC=1:4:2, weigh 125mg BSA, 500mg SUC and 250mg DEC respectively, BSA was pre-dissolved with anhydrous dimethylformamide (DMF), slowly add AA and DEC under stirring at room temperature , and stirred overnight at room temperature. The reaction solution was transferred to a dialysis bag and dialyzed in dist...

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Abstract

The invention discloses a synthesis method of an abietic acid artificial antigen. The synthesis method comprises the following steps: taking 12-hydroxyabietic acid (12-HA) as a hapten and carrying outtwo-step 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (DEC) catalysis reaction; taking adipic acid (AA) as a coupling arm to be coupled with keyhole limpet hemocyanin (KLH) and bovine serum albumin(BSA) respectively, so as to synthesize the abietic acid artificial antigens 12-HA-AA-KLH and 12-HA-AA-BSA respectively. A polyclonal antibody obtained by immunizing domestic rabbits through the abietic acid artificial antigen prepared by adopting the synthesis method has ideal valence and can be used for establishing an ELISA method for detecting rosin residues in livestock and poultry meat products.

Description

technical field [0001] The invention relates to a method for synthesizing an abietic acid artificial antigen, which belongs to the technical field of biochemistry. Background technique [0002] Rosin is a transparent solid substance obtained by evaporating turpentine from pine oleoresin. It is an important chemical raw material for making soap, paper, paint, etc. Abietic acid is the main component of rosin, which can cause asthma and skin allergies. Toxicological studies have shown that it can cause human alveolar epithelial cell lysis and has the activity of damaging DNA. [0003] Rosin has good adhesion properties after heating and melting, and has been widely used in the slaughter of waterfowl such as ducks and geese, as well as the secondary depilation processing of pig heads (claws). In 2009, my country promulgated the "Food Safety Law", prohibiting the use of rosin in the process of slaughtering and processing livestock and poultry, and recommending the use of its sub...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K14/795C07K1/107
CPCC07K14/765C07K14/795C07K19/00
Inventor 耿志明卞欢诸永志
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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