Amnesic shellfish toxin immunoaffinity column and preparation method and application thereof

A technology of shellfish toxins and immune affinity, applied in chemical instruments and methods, separation methods, other chemical processes, etc., can solve the phenomenon of poisoning and hypoxia, changes in blood properties, uncertain toxin components, and death of cultured shellfish To achieve the effect of ensuring antigen binding energy, improving antibody binding ability, and high specificity

Inactive Publication Date: 2018-07-27
山东美正生物科技有限公司
View PDF5 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Past studies believed that bivalve molluscs often have no adverse symptoms when they accumulate a large amount of DA, but with the deepening of research, it was found that red tide toxins have various sub-lethal or lethal effects on bivalve molluscs; for example, producing Paralytic Shellfish Toxins (PST) of Alexandrium tamari have effects on hatchability, survival rate, motility, filter-feeding rate and growth of bivalve molluscs, and can also inhibit shellfish closure motility, filter-feeding rate and filtration rate; Pseudonitzia P.multiserie containing ASP can cause respiratory acidosis and hypoxia of Pacific oyster Crassostreagigas and changes in blood properties; at the same time, toxic algal blooms may also cause a large number of farmed shellfish to die The main reason
[0004] Researchers have developed a variety of DA detection methods, including mouse biological detection, HPLLC-MS, fluorescence HPLC, GC-MS, TLC (thin layer chromatography), amino acid analysis, CE (capillary electrophoresis), CEC (Capillaryelectrochromatography), ELISA, receptor bindi

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Amnesic shellfish toxin immunoaffinity column and preparation method and application thereof
  • Amnesic shellfish toxin immunoaffinity column and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The main structure of the amnesic shellfish toxin immunoaffinity column involved in this embodiment includes a column body 1, a cover plate 2, a sample inlet 3, a first sieve plate 4, a packing material 5, a second sieve plate 6 and a sample outlet 7 The top of the column body 1 of circular tubular structure is provided with the cover plate 2 of circular sheet structure, and the center of cover plate 2 is provided with the sample inlet 3 of circular tubular structure, and the internal upper space of column body 1 is cavity, column The first sieve plate 4 with a circular sheet structure with a sieve hole is arranged in the middle of the interior of the body 1, the inner lower space of the cylinder 1 is filled with fillers 5, and the bottom of the cylinder 1 is provided with a circular plate with a sieve hole. The second sieve plate 6 with sheet-like structure is provided with a sample outlet 7 in a conical tubular structure with a wide top and a narrow bottom; the packing...

Embodiment 2

[0025] The specific process of the preparation method of the amnesic shellfish toxin immunoaffinity column involved in this example includes five steps of activation, coupling, connection, cross-linking and column packing:

[0026] (1) Activation: fully rinse the pre-swelled agarose gel sepharose4B with a concentration of 2% with distilled water, wash away the remaining ethanol in the agarose gel sepharose4B, the volume ratio of distilled water and agarose gel sepharose4B is 20:1, use Funnel filters liquid, obtains wet gel, takes by weighing 5 grams of wet gel and 7.5 milliliters of molar masses and is the NaOH (sodium hydroxide) aqueous solution of 0.8M, 2 milliliters of mass percentage concentrations and is 30% epichlorohydrin aqueous solution and 5 mL of NaBH at a concentration of 2 mg / mL 4 (Sodium borohydride) aqueous solution is carried out shaking table reaction under the condition of 25 ℃ for 8 hours at a temperature, after the reaction finishes, wash the wet gel with 5...

Embodiment 3

[0032] The specific process of the amnesic shellfish toxin immunoaffinity column purification and enrichment of the amnesic shellfish toxin involved in this embodiment is as follows: first, put 2.0g homogeneous shellfish meat Add 10 ml of 80% methanol aqueous solution to the stoppered centrifuge tube, vortex for 2 minutes, ultrasonically extract for 10 minutes, then centrifuge at 7000r / min for 5 minutes, and transfer the supernatant to No. 2 50 ml stoppered centrifuge tube In the No. 1 50 ml centrifuge tube with stopper, add 10 ml of 80% methanol aqueous solution by mass percentage, vortex for 2 min, then ultrasonically extract for 10 min, cool to room temperature, and then centrifuge at 7000 r / min for 5 min. Transfer the supernatant to No. 2 50ml centrifuge tube with stopper again, transfer 5mL supernatant to No. 3 50ml centrifuge tube, add 20ml PBS to dilute the supernatant; After equilibrating the amnesia shellfish toxin immunoaffinity column for 30 minutes, connect the inj...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention belongs to the technical field of food safety detection, and particularly relates to an amnesic shellfish toxin immunoaffinity column and a preparation method and application thereof. The preparation method includes: taking sepharose gel activated by adopting an epoxy chloropropane activation method as a carrier; coupling genetic recombinant protein G onto the carrier to obtain protein G-sepharose; connecting an amnesic shellfish toxin antibody onto the protein G-sepharose to obtain antibody-protein G-sepharose; using a crosslinking agent for crosslinking to obtain an antibody-protein G-spharose filler; packing the antibody-protein G-sepharose filler to form the amnesic shellfish toxin immunoaffinity column having high purity and affinity. The characteristic that the geneticrecombinant protein G is combined with the amnesic shellfish toxin antibody specifically is fully utilized, and Fab segment of the amnesic shellfish toxin antibody is exposed outside, so that antibodycombining capacity of the genetic recombinant protein G, capturing ability of amnesic shellfish toxin and purifying efficiency of the amnesic shellfish toxin are improved substantially.

Description

Technical field: [0001] The invention belongs to the technical field of food safety detection, and in particular relates to an amnesic shellfish toxin immunoaffinity column and a preparation method and application thereof. Background technique: [0002] The affinity column is a column installed after the substance that has a specific binding effect with the purification object is connected to an insoluble carrier to make an affinity adsorbent, and it is mostly used for affinity chromatography. [0003] The main component of Amnesic shellfish poisoning (ASP) is a physiologically active amino acid substance-Domoic acid (DA), which is produced by certain species of Pseudonitz and Nitzschia. Produced by diatoms; DA is an isomer of glutamic acid, a product of phytoplankton metabolism, an excitatory proline derivative and neurotoxin, an agonist of kainate receptors, capable of strongly binding Glutamate receptors act on excitatory amino acid receptors and synaptotransmitters, inc...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): B01J20/281B01J20/30B01D15/38G01N30/56G01N33/68G01N30/60
CPCB01D15/3809B01J20/281G01N30/56G01N30/60G01N33/68G01N2030/562
Inventor 柳家鹏王涛于蓉龚利新
Owner 山东美正生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap