Preparation method and application of neurotoxic shellfish poisoning immunoaffinity column

A shellfish toxin and immunophilic technology, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of reduced method application rate, low recovery rate, sample matrix interference, etc., to achieve rapid detection sensitivity, small matrix interference, and improved The effect of sample recovery

Pending Publication Date: 2020-06-19
品测(上海)检测科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, some solid-phase extraction column products on the market have low recovery rates and serious sample matrix interference, which greatly reduces the application rate of the method.

Method used

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  • Preparation method and application of neurotoxic shellfish poisoning immunoaffinity column
  • Preparation method and application of neurotoxic shellfish poisoning immunoaffinity column
  • Preparation method and application of neurotoxic shellfish poisoning immunoaffinity column

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: a kind of preparation method of neural shellfish toxin immunoaffinity column, comprising the following operation steps:

[0046] Step 1. Preparation of PbTX-2 monoclonal antibody:

[0047] Using the method of inducing monoclonal antibodies in animals, the whole antigen of PbTX-2 was synthesized to immunize mice, and then spleen cells of mice were fused with myeloma cells Sp2 / 0. Anti-PbTX-2-secreting monoclonal antibody cell lines were lethally injected into the abdominal cavity of mice by liquid paraffin, and then anti-PbTX-2-secreting cells were inoculated into mice to prepare anti-PbTX-2 monoclonal antibody ascites. After antibody purification using Protein G column, PbTX-2 monoclonal antibody was obtained.

[0048] The detailed operation is as follows:

[0049] 1. Preparation of complete antigen (ie whole antigen): The specific preparation method of complete antigen PbTX-BSA (used for immunization) is mainly to transform the hydroxyl group on the PbTX...

Embodiment 2

[0065] Example 2: A preparation method of a neuroshellfish toxin immunoaffinity column, which is different from Example 1 in that: in step 2, the measured protein concentration of the PbTX-2 monoclonal antibody solution is: 1 mg / mL .

Embodiment 3

[0066] Example 3: A preparation method of a neuroshellfish toxin immunoaffinity column, the difference from Example 1 is: in step 2, the measured protein concentration of the PbTX-2 monoclonal antibody solution is: 3 mg / mL .

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Abstract

The invention relates to a preparation method and application of a neurotoxic shellfish poisoning immunoaffinity column, which belong to the technical field of shellfish poisoning detection. The preparation method of the neurotoxic shellfish poisoning immunoaffinity column is technically characterized in that the NSP immunoaffinity column is prepared by purifying a PbTX-2 monoclonal antibody and taking agarose gel as a carrier. According to the application of the neurotoxic shellfish poisoning immunoaffinity column in neurotoxic shellfish poisoning detection, cultured and sea-caught mussels, clams, oysters and scallops are taken as samples, the samples are extracted with an aqueous solution containing 80% of methanol and are diluted with a PBS buffer solution, and after purifying and enriching with the NSP immunoaffinity column, and LC-MS / MS test analysis is carried out. According to the method, PbTX-2 is taken as a research object, column packing of the NSP immunoaffinity column is self-developed and prepared, a sample extracting solution purified by the immunoaffinity column can be directly used for LC-MS / MS analysis, and the purposes of rapid detection and improvement of the sensitivity and accuracy of the detection method are achieved.

Description

technical field [0001] The invention belongs to the technical field of shellfish toxin detection, and more particularly relates to a preparation method and application of a neuromuscular toxin immunoaffinity column. Background technique [0002] Shellfish toxin, also known as red tide toxin, is a natural active substance produced by organisms. Red tide is an ecological phenomenon in which protists, plankton or bacteria in seawater proliferate and cause discoloration in a specific environment. Among the more than 5,000 species of marine microalgae, there are at least 180 species of microalgae that can form red tides, of which nearly half are toxic algae, and 30-40 species have been confirmed to contain toxins or secrete toxins. Because researchers first discovered red tide toxins from shellfish, this toxin is called shellfish toxin. Red tide toxins are enriched in the body with the filter-feeding activities of filter-feeding fish, shrimp, and shellfish, causing seafood pois...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531
CPCG01N33/577G01N33/531
Inventor 董昕王佳杰戴辉朱亚琼周羽华董军
Owner 品测(上海)检测科技有限公司
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