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IgY+IgY(delta Fc)(H+L) and preparation thereof

A horseradish peroxidase and antibody-labeled technology, which is applied in the fields of analyzing materials through chemical reactions, material analysis and biological testing by observing the impact on chemical indicators, can solve the problem of enzyme-labeled antibodies hindering development, Insufficient and other problems, to achieve the effect of preventing various infectious diseases of geese

Inactive Publication Date: 2006-08-16
NORTHEAST AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

There are many reasons, among which the lack of a high-quality, commercially produced enzyme-labeled antibody is one of the main reasons hindering its development

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0025] 1) Purification of goose yolk immunoglobulin IgY and IgY(ΔFc)

[0026] According to v / v, the ratio of egg yolk: PBS (pH7.40.01mol / L): chloroform = 1:2:3 was mixed, centrifuged at 3000r / min for 30min, the supernatant was taken, and the final concentration (w / v) was 18 %, 14%, 9%, and 14% of anhydrous Na2SO4 were salted out in sequence, centrifuged at 3000r / min for 30min, and the supernatant was taken for the third time, and the precipitate was taken for the remaining three times. , centrifuge the dialyzate at 2000r / min for 10-15min, use PB (pH7.4, 0.01mol / L) as eluent, pass through DE 52 Cellulose column, 1ml / min, 1 tube / 5min, collect the first peak;

[0027] 2) Preparation and purification of rabbit anti-goose IgY+IgY(ΔFc)(H+L) antibody

[0028] For the first immunization, Freund's complete adjuvant and goose yolk immunoglobulin (0.4mg / ml) were mixed at a v / v ratio of 1:1, and 2ml / rabbit was injected subcutaneously in multiple points on the back; after that, booster i...

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Abstract

A method for preparing labeled antibody of rabbit resisting goose IgY+IgYú¿í¸Fcú®ú¿H+Lú®horseradish perxidase includes applying protein purification technique and enzyme-label technique of salting-out and chromatography ,immunizing rabbit for purifying and preparing rabbit resisting goose IgY+IgYú¿í¸Fcú®ú¿H+Lú®antibody based on purification of yolk immune globulin ,combining said antibody with horseradish peroxidase through modified sodium periodate ,utilizing the salting-out and sephodex G200 purification for obtaining antibody product .

Description

(1) Technical field [0001] The invention relates to an enzyme-antibody conjugate and a preparation method thereof, in particular to horseradish peroxidase-labeled rabbit anti-goose IgY+IgY(ΔFc)(H+L) antibody. (2) Background technology [0002] Enzyme immunoassay technology is a new type of detection technology based on the principle of enzyme reaction amplification and antigen-antibody specific binding characteristics, and based on enzyme-antibody and / or antigen conjugates. It includes enzyme immunohistochemical staining and enzyme immunoassay technology (including solid and non-solid phase enzyme immunoassay technology), the former is mainly used for tissue and cell immunoassay, and the latter is mainly used for antigen / antibody detection. [0003] Enzyme-linked immunosorbent assay (ELISA) is the fastest-growing and most widely used enzyme immunoassay technology, and it has gradually become a recognized detection technology due to its rapidity, accuracy and sensitivity. At...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/535G01N21/78
Inventor 王君伟孙凌霜
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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