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Method for preparing diarrhetic shellfish poisoning okadaic acid monoclonal antibody

A technology of okadaic acid and monoclonal antibody, which is applied in the field of marine biological pathogen research, can solve the problems of high price and the lack of widespread promotion of the kit, and achieve high specificity effect

Inactive Publication Date: 2015-07-29
GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

Due to the high price of foreign OA ELISA kits, domestically developed kits have not been widely promoted due to their own specificity and cross-reactivity.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0018] The specific embodiments of the invention will be further described in detail below.

[0019] 1. Preparation of immune antigen (OA-IgG) and detection antigen (OA-BSA)

[0020] 1mgOA, 0.155mg N-hydroxysuccinimide, 0.285mg N, N dicyclohexanecarbodiimide were mixed in 60μL DMF, and incubated at room temperature for 2h; 20μL reaction solution was added with 0.75mg IgG (dissolved in 50μL 0.1mol / L NaHCO 3 ), add 0.95mg BSA (dissolved in 50μL 0.1mol / L NaHCO 3 ), continued incubation at room temperature for 2 h; unreacted substances were removed by ultrafiltration and centrifugation, and the coupled antigen was dissolved with an appropriate volume of PBS (pH 7.4) to a concentration of 1 g / L, and stored at -20°C for future use.

[0021] The invention adopts the active ester method to couple the small molecular toxin OA to the macromolecular carrier protein IgG and BSA through N-hydroxysuccinimide as an intermediate under the action of DCC. OA is insoluble in water, and the c...

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PUM

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Abstract

The invention provides a method for preparing a diarrhetic shellfish poisoning okadaic acid monoclonal antibody. The method comprises the following steps: firstly, preparing an okadaic acid complete antigen; obtaining a hybridoma cell strain stably secreting an anti-OA (okadaic acid) antibody; finally preparing a large amount of monoclonal antibodies. According to the method provided by the invention, the preparation process is optimized, so that large-batch preparation of the diarrhetic shellfish poisoning okadaic acid monoclonal antibody becomes possible, and the foundation is provided for further developing an OA ELISA (enzyme-linked immuno sorbent assay) rapid detection kit which is high in specificity, free of cross reaction, sensitive and low in cost.

Description

technical field [0001] The invention belongs to the technical field of marine biological pathogen research, and in particular relates to a preparation method of a monoclonal antibody to okadaic acid, a diarrheal shellfish toxin. Background technique [0002] Diarrhetic Shellfish Poisoning (DSP) is a class of fat-soluble secondary metabolites produced by marine algae or microorganisms. Because filter-feeding shellfish ingest these algae and microorganisms that produce DSP toxins, the toxins can accumulate in the body, which will cause symptoms such as severe diarrhea, vomiting, and lower abdominal pain in the eaters, and even endanger life. This type of toxin is stable to heat, and there is no specific antidote at present, and the treatment of patients is generally carried out by symptomatic treatment. Therefore, strengthening the detection of such toxins has become the only measure to effectively prevent the occurrence of food poisoning incidents. [0003] At present, the ...

Claims

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Application Information

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IPC IPC(8): C07K16/14
Inventor 刘义王本成卢虹玉李延平
Owner GUANGDONG MEDICAL UNIV
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