Preparation method of colloidal carbon labeled antibody and test strip prepared from colloidal carbon labeled antibody

A technology for labeling antibodies and colloidal carbon, which is applied in the preparation of colloidal carbon-labeled antibodies and test strips, can solve the problems of large batch-to-batch differences, high costs, and weak signal strength, and achieve less antibody consumption, cost savings, and lower cost effect

Inactive Publication Date: 2015-08-19
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

[0006] In view of this, the present invention aims to propose a method for preparing a colloidal carbon-labeled antibody and an immunochromatographic analysis method, which can quickly and visually detect salbutamol residues, so as to solve the problem of high cost of the colloidal gold-labeled immunochromatographic analysis method and large batch-to-batch differences. , weak signal strength, etc.

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  • Preparation method of colloidal carbon labeled antibody and test strip prepared from colloidal carbon labeled antibody
  • Preparation method of colloidal carbon labeled antibody and test strip prepared from colloidal carbon labeled antibody
  • Preparation method of colloidal carbon labeled antibody and test strip prepared from colloidal carbon labeled antibody

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Embodiment 1

[0036] A method for preparing a colloidal carbon-labeled antibody, comprising the following steps,

[0037] 1) Disperse the carbon black in the buffer solution, and ultrasonicate for 20 minutes in an ice bath to obtain a uniformly dispersed colloidal carbon solution;

[0038] 2) Dilute the albuterol antibody to 0.5mg mL with buffer -1 ;

[0039] 3) Add the albuterol antibody to the colloidal carbon solution at a stirring speed of 100 rpm, and stir for 12 hours at 4° C. at a stirring speed of 100 rpm to obtain a colloidal carbon-labeled antibody;

[0040] 4) Wash the colloidal carbon-labeled antibody with washing buffer, repeat 3 times, centrifuge at 12000r / min, 10°C for 30min; resuspend the final pellet with borate buffer to the original volume, and store at 4°C for later use.

[0041] Wherein, in step 1) and step 2), the buffer is borate buffer, and its pH is 8.8. In step 3), the concentration of albuterol antibody is 80 μg mL -1 ;The concentration of colloidal carbon is ...

Embodiment 2

[0056] A method for preparing a colloidal carbon-labeled antibody, comprising the following steps,

[0057] 1) Disperse the carbon black in the buffer solution, and ultrasonicate for 30 minutes in an ice bath to obtain a uniformly dispersed colloidal carbon solution;

[0058] 2) Dilute the albuterol antibody to 0.8mg mL with buffer -1 ;

[0059] 3) Add the albuterol antibody to the colloidal carbon solution at a stirring speed of 100 rpm, and stir at 25°C for 12 hours at a stirring speed of 100 rpm to obtain a colloidal carbon-labeled antibody;

[0060] 4) Wash the colloidal carbon-labeled antibody with washing buffer, repeat twice, and centrifuge at 10,000 r / min, 4°C for 20 min; resuspend the final pellet with borate buffer to the original volume, and store at 4°C for later use.

[0061] Wherein, in step 1) and step 2), the buffer is borate buffer, and its pH is 8.8. In step 3), the concentration of albuterol antibody is 120 μg mL -1 ;The concentration of colloidal carbon...

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Abstract

The invention provides a preparation method of a colloidal carbon labeled antibody and a test strip prepared from the colloidal carbon labeled antibody. Salbutamol is selected as a target object, a salbutamol antibody is added into a colloidal carbon solution to prepare the colloidal carbon labeled antibody, and a colloidal carbon labeled immunochromatography analysis method for detecting the salbutamol is established and is applied to visual detection of salbutamol residue in animal derived food. The detection limit of the prepared test strip is 2 mu g L<-1>. Four practical samples namely a pork sample, a beef sample, a mutton sample and a pork liver sample are selected to perform addition recovery experiment, and the detection limit of the samples is 10 mu g L<-1>. The test strip can be stably stored for at least 16 weeks at room temperature.

Description

technical field [0001] The invention belongs to the field of rapid detection of veterinary drug residues, and in particular relates to a preparation method of a colloidal carbon-labeled antibody and a test strip prepared by using the same. Background technique [0002] The immunochromatographic analysis method has the advantages of high sensitivity, simple operation, time-saving and labor-saving, low cost, and is suitable for rapid on-site screening of a large number of samples. It has developed rapidly in the field of veterinary drug residue detection in recent years. Colloidal gold has the advantages of good biocompatibility, different particle sizes can show different colors, and it is easy to improve the signal intensity through modification. It has become the most commonly used labeling material in immunochromatographic analysis methods. However, colloidal gold is expensive and has large batch-to-batch differences, which is not conducive to the mass production of test s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/532
CPCG01N33/558G01N33/532
Inventor 刘冰王玲玲张燕生威王硕
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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