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Penicillin and carrier protein couplet product, method for producing beta-lactam penicillin antibody, and uses thereof

A carrier protein, lactam technology, applied in biochemical equipment and methods, chemical instruments and methods, ovalbumin and other directions, can solve the problem of high price, and achieve accurate and reliable results, strong specificity, simple and convenient operation. Effect

Inactive Publication Date: 2009-05-13
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Some foreign manufacturers have developed ELISA detection kits for some drug residues, but they are expensive

Method used

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  • Penicillin and carrier protein couplet product, method for producing beta-lactam penicillin antibody, and uses thereof

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Embodiment Construction

[0032] The invention provides methods and uses for preparing penicillin and carrier protein coupling products and beta-lactam penicillin antibodies. The prepared immunogen is used to prepare β-lactam antibiotics that have strong specificity, high sensitivity, good stability, and can be mass-produced monoclonal antibodies and polyclonal antibodies, and use these antibodies to establish the detection of β-lactam antibiotic residues in food A rapid diagnostic method with high specificity, sensitivity and accuracy-competitive ELISA and immunochromatographic test strips, can be effectively used for the detection of β-lactam antibiotic residues in animal products, serving my country's food safety.

[0033] 1. The steps of a preparation method of penicillin and bovine serum albumin (BSA) coupling product:

[0034] a) Add 20 mg of penicillin sodium salt, 5 mg of N-hydroxysuccinimide and 20 mg of carbodiimide to 1 ml of ultrapure water, and stir for 10-50 minutes in an ice bath;

[0035] b...

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Abstract

The invention provides a preparation method and application of a product obtained by coupling penicillin with carrier protein, as well as a beta-lactam type penicillin antibody. Animals are immunized with penicillin artificial antigen coupled in the invention so as to prepare the antibody which can be used for detecting beta-lactam type penicillin in foods. The preparation method comprises the following steps: immune BALB / C mouse spleen cells and SP2 / 0 mouse myeloma cells are fused; beta-lactam type antibiotics coupled with the carrier protein are used as coating antigen to screen positive hybridoma; hybridoma capable of stably transferring culture and secreting anti-beta-lactam type antibiotic antibodies through cell clones is obtained; and an ascites monoclonal antibody is prepared. The prepared monoclonal antibody is utilized to establish a direct competitive ELISA method having high specificity, sensitivity and accuracy to the beta-lactam type antibiotics, as well as an immune colloidal gold test strip. The preparation method for the product obtained by coupling penicillin with carrier protein, as well as the beta-lactam type antibiotic antibodies can serve the rapid detection of beta-lactam type antibiotic residue in foods.

Description

Technical field [0001] The invention belongs to the field of immunochemical biology technology, and in particular relates to a preparation method and application of a penicillin and carrier protein coupling product and a beta-lactam penicillin antibody. Background technique [0002] Penicillin is an important class of β-lactam antibiotics. They can be extracted from fermentation broth or semi-synthesized. The mechanism of action of various β-lactam antibiotics is similar. Lactam antibiotics are similar to the β-lactam on the cell membrane. The amide antibiotic binding protein (PBP) binds to hinder the synthesis of bacterial cell wall mucopeptides, so that they cannot be cross-linked and cause cell wall defects, causing bacterial cells to rupture and die. This process occurs during the reproduction period of bacterial cells, so this class of drugs is a bactericide during the reproduction period. [0003] β-lactam antibiotics play an antibacterial effect by inhibiting the activity ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/795C07K14/765C07K14/77C07K16/00C07K16/44C07K16/06C12N9/00G01N33/53G01N33/577
Inventor 吴建祥
Owner ZHEJIANG UNIV
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