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Dual-specificity monoclonal antibody for resisting chloramphenicol and apramycin and preparation method thereof

A monoclonal antibody, apramycin technology, applied in chemical instruments and methods, specific peptides, hybrid immunoglobulins, etc., can solve problems such as cumbersome processing methods, low sensitivity, and unsuitable batch sample screening and detection tasks. , to achieve the effect of stable character and high detection sensitivity

Inactive Publication Date: 2013-05-08
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method is simple in principle and easy to operate, the time required for the determination is too long and the sensitivity is not high.
Although the results of physical and chemical detection methods are accurate, the detection equipment is expensive and requires professional operation. development trend
[0011] At present, although there are reports on the immunoassay techniques of chloramphenicol and apramycin, they are all limited to a single analysis technique and method. In the context of the need for multi-residue analysis, it is necessary to establish an immunoassay that can simultaneously detect two or two types of target compounds. Immunoassay technology is particularly urgent, and the preparation of high-quality bispecific monoclonal antibodies is a new means and method for establishing immunological multi-residue analysis technology

Method used

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Embodiment Construction

[0022] The present invention will be further described below in conjunction with the examples.

[0023] 1. Preparation of Immunogen

[0024] 1.1 Preparation of chloramphenicol hapten, artificial antigen and coated antigen

[0025] 1.1.1 Synthesis of chloramphenicol succinate

[0026] Weigh 2.0 g of chloramphenicol and 0.75 g of succinic anhydride, place them in a 100 mL three-neck flask, connect the reflux condenser to a calcium chloride drying tube or vacuumize and fill with nitrogen to keep the reaction process anhydrous and oxygen-free. Inject 15-30 mL of anhydrous and oxygen-free pyridine, stir to dissolve, and react at 60-90 °C for 2-12 h. After the reaction solution (dark rose red) is naturally cooled to room temperature, add 50 mL of water and use 6 mol / Adjust the pH of the mixture to 5 with L hydrochloric acid, extract 3 times with 30 mL of ethyl acetate, extract the organic phase with 3 × 25 mL of 1 mol / L aqueous sodium bicarbonate solution, collect the aqueous pha...

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Abstract

The invention discloses a dual-specificity monoclonal antibody for resisting chloramphenicol and apramycin as well as a preparation method and application thereof, belonging to the technical field of biotechnology. The dual-specificity monoclonal antibody simultaneously has specificity for the chloramphenicol and the apramycin. The preparation method of the dual-specificity monoclonal antibody comprises the steps of: synthesizing haptens and artificial antigens of the chloramphenicol and the apramycin; preparing and secreting chloramphenicol resistance hybridoma cell strains by using the artificial antigens of the chloramphenicol; preparing and secreting apramycin resistance hybridoma cell strains by using artificial antigens of the apramycin; fusing the chloramphenicol resistance hybridoma cell strains and the apramycin resistance hybridoma cell strains, then screening to obtain tetrasomic hybridoma cells which secrete the dual-specificity monoclonal antibody for resisting chloramphenicol and apramycin. The dual-specificity monoclonal antibody obtained by the method has stable character and high detection sensitivity and can be produced in large quantities; and an ELISA (Enzyme Linked Immunosorbent Assay) analytical method is established by using the antibody and provides technological base for rapid detection of the chloramphenicol and apramycin.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a bispecific monoclonal antibody against chloramphenicol and apramycin, a preparation method and application thereof. Background technique [0002] Immunoassay (IA) is a technology that uses antibodies as biochemical detectors to perform qualitative and quantitative analysis of compounds, enzymes, or proteins. It is based on the specific and reversible binding reaction between antigens and antibodies. It has the characteristics of strong specificity, high sensitivity, convenience and speed, high throughput, low detection cost, safety and reliability. This method generally does not require expensive instruments, and the technical requirements of the users are not high. It is easy to popularize and promote, and is especially suitable for on-site screening and rapid analysis of a large number of samples. A series of detection products developed based on this technology, suc...

Claims

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Application Information

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IPC IPC(8): C07K16/46C07K16/44
Inventor 金仁耀
Owner ZHEJIANG GONGSHANG UNIVERSITY
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