Unlock instant, AI-driven research and patent intelligence for your innovation.

Preparation method of valnemulin egg yolk antibody

A Vonimulin and egg yolk antibody technology, which is applied in the field of antibody preparation, can solve the problems such as no reports on the preparation of Vonimulin egg yolk antibody, few applications of egg yolk antibody, etc., and achieves good antibody specificity, good application prospect, and yield. high effect

Inactive Publication Date: 2015-01-14
CHINA AGRI UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, yolk antibodies are rarely used in the field of detection of veterinary drug residues, and there is no report on the preparation of yolk antibodies to warnimulin

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
  • Preparation method of valnemulin egg yolk antibody
  • Preparation method of valnemulin egg yolk antibody
  • Preparation method of valnemulin egg yolk antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, the preparation of warnimulin egg yolk antibody

[0056] 1. Antigen synthesis of warnemulin

[0057] Dissolve 100mg of carrier protein bovine serum albumin (BSA) and 50mg of warnemulin hydrochloride in 2mL of 0.01M (pH7.4) PBS buffer to obtain solution I; weigh 100mg of carbodiimide (EDC), dissolve In 1mL of pure water, the solution II was obtained; the solution II was added to the solution I, and the reaction was stirred at 20-25°C for 2-3h to obtain the target product VAL-BSA, which was 0.01mol at room temperature (20-25°C) / L PBS solution for 3-4 days and stored at -20°C.

[0058] 2. Immunization of laying hens with warnimulin immunogen

[0059] The laying hens were immunized with the immunogen VAL-BSA.

[0060] For the first immunization, VAL-BSA (dissolved in 0.01mol / L PBS solution) was emulsified with an equal volume of Freund's complete adjuvant, and then multi-point immunization was carried out on the back of laying hens. The immunization dose o...

Embodiment 2

[0067] Embodiment 2, the identification of warnimulin egg yolk antibody

[0068] 1. Determination of antibody production

[0069] The concentration of the yolk antibody of Wernemulin was detected by NaNoDrop spectrophotometer, and the results showed that the average concentration of the yolk antibody of Wernemulin was about 15-18mg / mL, that is, about 1g of yolk antibody could be collected from each egg yolk, and after the first immunization The amount of yolk antibody collected in the whole test period (43 weeks) from the beginning to the end of the test was about 20 g.

[0070] 2. Determination of antibody titer

[0071] The method of step 1 in Example 1 was used to prepare the original coating VAL-OVA, wherein only ovalbumin was used instead of bovine serum albumin.

[0072] Indirect ELISA to measure the titer of yolk antibody of warnemulin, the method is as follows:

[0073] (1) Coating: Dilute the coated original VAL-OVA to 0.6 μg / mL with 0.05 M carbonate buffer (pH 9.6...

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 discloses a preparation method of a valnemulin egg yolk antibody. The invention discloses a method for preparing the valnemulin egg yolk antibody. The method comprises the following steps: coupling valnemulin hydrochloride with a carrier protein by virtue of carbodiimide to obtain an immunogen; immunizing a laying hen by using the immunogen to obtain an egg of the immunized laying hen; and extracting the valnemulin egg yolk antibody from the egg yolk. The valnemulin egg yolk antibody prepared by the method disclosed by the invention has the unique advantages of being simple and economic in preparation process, high in output and the like, and has a good application prospect in detection of veterinary drug residues.

Description

technical field [0001] The invention relates to a preparation method of a warnemulin antibody; in particular, it relates to a preparation method of a warnemulin yolk antibody, which belongs to the field of antibody preparation. Background technique [0002] Valnemulin (VAL) is a new generation of Pleuromutilin semi-synthetic antibiotics, which has attracted widespread attention due to its high safety, broad antibacterial spectrum, strong antibacterial activity, and low drug resistance. The drug is mainly used for the prevention and treatment of mycoplasma and intestinal spirochete infection in livestock and poultry. However, due to its wide application, its residual phenomenon in animal tissues is relatively serious, which may cause some adverse reactions. In addition, warnemulin can also interact with polyether drugs to cause animal poisoning. Therefore, the European Medicines Evaluation Agency (EMEA) stipulates that the maximum residue limit (MRL) of warnemulin in pig tis...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/02C07K1/30G01N33/53
Inventor 王战辉沈建忠史为民张素霞江海洋温凯张会艳盛雅洁
Owner CHINA AGRI UNIV