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Preparation method of anti-microcystin LR egg yolk antibodies

A technology of microcystin and yolk antibody, which is applied in chemical instruments and methods, antibacterial immunoglobulins, material testing products, etc., can solve the problems of uninvolved formulation research, weak basic research work, etc., and achieve good application prospects. , the effect of good economic and social benefits

Inactive Publication Date: 2017-04-26
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the basic research work on the application of IgY technology is still relatively weak, and almost no one has been involved in the research on its dosage form, which has also become a "bottleneck" for the general application of IgY

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  • Preparation method of anti-microcystin LR egg yolk antibodies

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Embodiment 1

[0040] (1) Dissolve 3 mg microcystin in 1.2 mL dimethylformamide (DMF), add 100 mL activation solution (12 mg EDC, 0.1 mol / L HCl and 7 mg NHS dissolved in 1 mL DMF), and activate at 4°C overnight. Take 20 mg KLH in 5 mL pH9.6 carbonic acid buffer, stir and place in an ice-water bath, add the above-mentioned activated microcystin drop by drop, continue stirring for 1 h after the addition, and then place it at 4°C overnight. Dialyze with 0.01M PBS for 3 days, centrifuge to get the supernatant, and use the lorry method to determine the concentration to be 1.4 mg·mL –1 , stored at -20°C, and the obtained MC-LR-KLH was used as an immunizing antigen.

[0041] (2) MC-LR-KLH and Fisher's complete adjuvant were mixed in a volume ratio of 1:1 as immunogen 1; MC-LR-KLH and Fisher's incomplete adjuvant were mixed in a volume ratio of 1:1 as Immunogen 2; MC-LR-KLH (volume ratio 1:1) was dissolved in 0.01M PBS as immunogen 3.

[0042] The 28-week-old Rohman layer hens were collected from ...

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Abstract

The invention relates to a preparation method of anti-microcystin LR egg yolk antibodies. The preparation method includes: coupling microcystin LR with hemocyanin KLH to obtain microcystin LR-hemocyanin MC-LR-KLH; using the MC-LR-KLH as the immunizing antigen to immunize laying hens to obtain anti-microcystin-LR immunized eggs; breaking the anti-microcystin-LR immunized eggs, taking the yolk parts, purifying, and performing freeze drying to obtain the anti-microcystin LR egg yolk antibodies. The antibodies prepared by the method can be used for detecting the microcystin LR and are good in economic and social benefits and promising in application prospect.

Description

technical field [0001] The invention belongs to the field of preparation of egg yolk antibody, in particular to a preparation method of anti-microcystin LR egg yolk antibody. Background technique [0002] Microcystin (MC) is a class of secondary metabolites produced by cyanobacteria. It belongs to cellular endotoxins. It is released into water after cell death or when the cell membrane permeability is enhanced. The system is easy to cause serious harm, and drinking water and aquatic products contaminated by microcystins have brought a huge threat to human health. [0003] Globally, microcystins consisting of seven amino acids are the most common cyanotoxins in freshwater environments and have a ring structure (-D-Ala-L-X-D-Masp-L-Z-Adda-D-Glu-Mdha) At present, there are at least 80 variants of microcystins, all of which have a molecular weight of about 1 000. Among them, three combinations of algae toxins are the most toxic: LR, RR, and YR, respectively, where L, R, and Y r...

Claims

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Application Information

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IPC IPC(8): C07K16/12C07K16/02G01N33/68
CPCC07K16/12C07K16/02C07K2317/35G01N33/68G01N2333/195G01N2410/00
Inventor 曹军朱春锋
Owner NANTONG UNIVERSITY