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Protein chip for detecting sugar chain abnormal IgA kidney disease

A protein chip and abnormal technology, applied in the field of protein chips, can solve the problems of not being able to adapt to large-scale screening of high-incidence population, complicated device use, etc., and achieve the effects of shortened detection time, low cost, and high upgrade potential

Active Publication Date: 2014-05-07
上海睿昂基因科技股份有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the device of this invention is relatively complicated to use, and cannot meet the needs of large-scale screening of high-risk groups

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Detection of Abnormal Sugar Chain IgA Nephritis by Protein Chip

[0038] 1. Snail agglutinin (HAA / HPA) and chip immobilization

[0039] The chip is made of polycarbonate material. The snail lectin HAA is dissolved in the crystal chip sampling solution. The concentration of the snail lectin is 1.0mg / ml. The sample diameter is 50-100 microns; after spotting, place the chip at room temperature overnight.

[0040] 2. Processing samples

[0041] Take 400 parts of serum from patients with abnormal sugar chain IgA nephritis and 600 parts of normal human serum, and dilute with 0.1M PBS buffer solution with pH 7.4;

[0042]3. Use YL-JCY2.0 protein chip detector for detection

[0043] 1) Take the protein chip and add 100 μl 0.1M Tris-HCl to react at room temperature for 1.5 hours to block, wash the chip with 0.1M Tris-HCl-Tween20 solution 3 times, add reference substance, negative control substance, positive control substance and test sample, React at room temperatu...

Embodiment 2

[0048] Example 2 Detection of Abnormal Sugar Chain IgA Nephritis Using Protein Chip

[0049] 1. Immobilization of villous vetch lectin (VVL) to the chip

[0050] The chip is made of polycarbonate material, and the snail lectin HAA is dissolved in the crystal core chip sample solution, and the sample concentration of villous vetch lectin is 1.5 mg / ml; the lectin is sampled on the chip with a spotting instrument , the spotting volume is 40nl, and the spotting diameter is 100-150 microns; after spotting, leave the chip at room temperature overnight.

[0051] 2. Processing samples

[0052] Take 400 serums from patients with abnormal sugar chain IgA nephritis and 600 serums from normal people, and dilute them with 0.1M PBS buffer;

[0053] 3. Use YL-JCY2.0 protein chip detector for detection

[0054] 1) Take the protein chip and add 100 μl of 0.1M Tris-HCl to react at room temperature for 1.5 hours to block, wash the chip with 0.1% TBS-Tween20 solution for 3 times, add reference...

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Abstract

The invention relates to the field of protein chips, in particular to a protein chip for detecting a sugar chain abnormal IgA kidney disease. The protein chip is prepared by fixing lectin on a solid vector and using a marked second antibody to be combined with IgA in a detection sample, wherein the lectin is selected from snail lectin or vicia villosa lectin. The protein chip provided by the invention is specifically combined with special sugar chain protein by using the lectin so as to diagnose the disease, thereby providing a new way for the diagnose of the disease. The method is simple and practicable, the cost is low, the operation steps are simplified, the detection time is shortened, and the method is a novel method which is appropriate to generalize.

Description

technical field [0001] The invention relates to the field of protein chips, in particular to a protein chip for detecting IgA nephropathy with abnormal sugar chains and a detection method for IgA nephropathy with abnormal sugar chains. Background technique [0002] The protein chip is a combination of planar microfabrication and biomolecular self-assembly technology, and assembles a large number of different biomolecular microarrays on tiny carriers to achieve large-scale detection of target molecular information. [0003] Glycobiology is a science that studies the structure, chemistry, biosynthesis and biological functions of glycans and their derivatives. Scientists call the science of studying polysaccharides in organisms "glycobiology", and some people follow the concepts of "genomics" and "proteomics" to call this discipline "glyconomics". Proteins, nucleic acids and polysaccharides are the three types of macromolecules that constitute life. The research on proteins an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543
Inventor 刘睿强
Owner 上海睿昂基因科技股份有限公司
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