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Yolk immunoglobulin detection method

A technology of egg yolk immunoglobulin and detection method, which is applied in biological testing, measuring devices, material inspection products, etc., and can solve the problems of inaccurate analysis of purity, specificity and content, poor repeatability, and low detection accuracy

Inactive Publication Date: 2019-11-19
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the existing IgY detection method, only the biological activity of IgY can be qualitatively determined, but its purity, specificity and content cannot be accurately analyzed, and there are deficiencies such as low detection accuracy and poor repeatability

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Embodiment 1

[0046] The detection method of the IgY of the present embodiment comprises the following steps:

[0047] 1. Preparation of protein samples.

[0048]Protein samples with known protein concentrations were mixed with 5×loading buffer at a ratio of 1:4 by volume, placed in a metal bath at 95°C for 5 minutes for denaturation, and then stored in a -80°C refrigerator until use. The purpose of this step is to open the spatial structure of the protein and allow the buffer to fully bind to it.

[0049] 2. With glue.

[0050] Configure the separating gel and stacking gel according to the needs (such as the instructions of the existing gel dispensing kit). This step is intended to: prepare for subsequent gel electrophoresis.

[0051] 3. Sample loading.

[0052] Clean the glass plate (to avoid the influence of residual reagents in the experiment), check for leaks, and use the configured stacking gel to fill the gel. After the stacking gel solidifies, transfer the gel plate to the elect...

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Abstract

The present invention relates to a yolk immunoglobulin detection method, comprising the following steps: (1) performing glue filling on prepared spacer gel, and separately adding a denatured protein sample with a known concentration into multiple sample loading holes; (2) performing electrophoresis on the protein sample that undergoes sample loading; (3) placing entire gel that undergoes electrophoresis on an activated solid-phase holder, and then transferring the entire gel to a transfer buffer with ice blocks, to transfer membrane; (4) sealing the solid-phase holder that undergoes membrane transferring; (5) cleaning the sealed solid-phase holder, and then performing antibody incubation by using an HRP-containing IgY antibody; (6) cleaning the solid-phase holder that undergoes the antibody incubation, then performing color development by using an ECL coloring solution, and performing image development by using a chemiluminiscence visualizer; and (7) performing gray analysis on an image obtained through image development. According to the IgY detection method provided in the present invention, not only an IgY content can be detected, but also IgY purity can be analyzed.

Description

technical field [0001] The invention relates to an immunoglobulin inspection technology, in particular to a method for detecting egg yolk immunoglobulin (IgY). Background technique [0002] Yolk immunoglobulin (Immunoglobulin of Yolk, IgY) is the only immunoglobulin in yolk, commonly found in birds, reptiles and amphibians, and functionally equivalent to mammalian immunoglobulin IgG. IgY produces quickly, has a large output, high potency, stable physical and chemical properties, wide sources, and no side effects, so it is a good raw material for special medical food. At present, the extraction methods of IgY include water dilution method, ammonium sulfate extraction, polyethylene glycol method, dextran sulfate method, xanthan gum method, cold ethanol fractionation method and water dilution ultrafiltration method, etc. Generally, after IgY is extracted, it is purified by gel chromatography or ion exchange chromatography. The more commonly used IgY detection method is ELISA ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N21/76G01N1/34G01N1/40
CPCG01N33/6854G01N33/543G01N21/76G01N1/34G01N1/40G01N2001/4038G01N2600/00
Inventor 张南文黄丽珊闫梦茹刘蕊张鑫于鹤陈智婕陈晓乐
Owner FUJIAN MEDICAL UNIV