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Preparation and application of a penicillin-tremorin binding antigen and its antibody

A penicillin-coated technology, applied in the field of biomedicine, can solve the problems of expensive equipment, unsuitable for rapid analysis and determination of a large number of samples, cumbersome processing steps, etc., and achieves the effect of high price

Active Publication Date: 2019-03-08
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the detection of penicillin tremor is mainly carried out by instrumental analysis methods, including LC-UV, LC-MS and LC-MS-MS analysis methods, but these instrumental analysis methods have expensive equipment and high technical requirements for staff. It is not suitable for rapid analysis and determination of a large number of samples due to its shortcomings such as cumbersome pre-processing steps

Method used

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  • Preparation and application of a penicillin-tremorin binding antigen and its antibody
  • Preparation and application of a penicillin-tremorin binding antigen and its antibody
  • Preparation and application of a penicillin-tremorin binding antigen and its antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1, the preparation of the enzyme-linked immunosorbent assay kit that detects penicillin tremor

[0053] 1. Preparation of penicillin-tremorin conjugated antigen

[0054] The penicillin-tremorin-binding antigen is prepared according to the method comprising the following steps: the penicillin-tremorin-binding antigen is obtained by coupling the penicillin-tremorin to a carrier protein, and the carrier protein is hemocyanin (KLH) or bovine serum albumin (BSA) , the coupling is achieved through a Mannich reaction. The specific method is as follows:

[0055] Dissolve 10 mg of hemocyanin or 15 mg of bovine serum albumin (BSA) in 2.0 mL of sodium acetate solution with a concentration of 0.1 mM and a pH value of 4.2 to prepare a sodium acetate solution of hemocyanin with a concentration of 5 mg / mL Or a sodium acetate solution of bovine serum albumin with a concentration of 7.5 mg / mL. 2.0 mg of penicillin tremor ( figure 1 ) was dissolved in 0.6mL solvent to prep...

Embodiment 2

[0075] Embodiment 2, the sensitivity and the specificity test of the ELISA kit of penicill tremor

[0076] 1. Sensitivity test of penicillin-tremorine enzyme-linked immunosorbent assay kit

[0077] The penicillin tremor standard was diluted with coating buffer to prepare penicillin standard solutions with concentrations of 0, 3, 9, 27, 81, 243 and 729 ng / mL as the experimental solution. Six groups of parallel experiments (n=6) were used.

[0078] Determine the working concentration of the penicillin tremorin polyclonal antibody prepared by embodiment 1 and the penicillin tremorin polyclonal antibody prepared by the square matrix titration method, and the optimal dilution factor of the penicillin tremorin coater is 1: 1000, the best dilution factor of penicillin-tremorin polyclonal antibody is 1:1000.

[0079] Conduct the following experiments:

[0080] 1. Coating: dilute the penicillin tremorin coating original prepared in Example 1 with a coating buffer to obtain a penicil...

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Abstract

The invention discloses a penitrem A combined antigen as well as preparation and an application of an antibody of the antigen. The median inhibitory dose (IC50), detected by a penitrem A enzyme-linked immunosorbent assay kit, of penitrem A for a reaction between a specific antibody of the penitrem A and penitrem A coating antigens is 100 ng / mL, the lowest detection limit of penitrem A is 30 ng / mL, and the cross reaction rate of penitrem A and an aflatoxin B1 sample, the cross reaction rate of penitrem A and a cyclopiazonic acid sample, the cross reaction rate of penitrem A and a vomitoxin and the cross reaction rate of penitrem A and a citreoviridin sample are all smaller than 0.1%. The specific antibody of penitrem A in the penitrem A enzyme-linked immunosorbent assay kit has the characteristics of high potency, high sensitivity and high specificity and can be used for quickly, sensitively and easily detecting the residual amount of penitrem A in animal-origin food.

Description

technical field [0001] The invention relates to the preparation and application of a penicillin-tremorin binding antigen and its antibody in the field of biomedicine. Background technique [0002] Penitrem A, a fungal neurotoxin produced by certain Aspergillus, Claviceps and Penicillium species, can inhibit smooth muscle potassium channels, It was isolated for the first time from moldy feed that caused neurological symptoms in sheep. It mainly injures the central nervous system, causing tremors, convulsions, and even death in animals. [0003] At present, the detection of penicillin tremor is mainly carried out by instrumental analysis methods, including LC-UV, LC-MS and LC-MS-MS analysis methods, but these instrumental analysis methods have expensive equipment and high technical requirements for staff. Due to its shortcomings such as cumbersome pre-processing steps, it is not suitable for rapid analysis and determination of a large number of samples. The immunoassay detec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/531C07K14/795C07K14/765C07K14/77C07K14/47C07K16/14
CPCC07K14/47C07K14/765C07K14/77C07K14/795C07K16/14C07K19/00G01N33/531G01N33/54353
Inventor 王战辉张素霞史为民曹明慧杨慧娟
Owner CHINA AGRI UNIV
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