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System for separating protein polypeptide in blood serum and reagent kit thereof

A serum and buffer technology, applied in the analysis of materials, material analysis by electromagnetic means, measurement devices, etc., can solve the problems that cannot meet the requirements of broad-spectrum adsorption and specific adsorption, and achieve convenient operation, low cost, and sample use. less effect

Inactive Publication Date: 2010-10-13
BIOYONG TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Although the system can realize the adsorption and resolution of specific protein polypeptides by magnetic beads, it still cannot meet the requirements of broad-spectrum adsorption and specific adsorption of magnetic beads to protein polypeptides in serum at the same time.

Method used

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  • System for separating protein polypeptide in blood serum and reagent kit thereof
  • System for separating protein polypeptide in blood serum and reagent kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1: the preparation of SPE-C system of the present invention and SPE-C kit thereof

[0048] According to the magnetic bead system for separating protein and polypeptide, prepare eluent, equilibration buffer, binding buffer and washing buffer to obtain systems A-C.

[0049] combination

equilibration buffer

0.25M, pH6.0

binding buffer

0.15M, pH5.6

wash buffer

0.35M, pH5.0

System A

Citrate

Phthalate

Citrate

System B

Acetate

Acetate

Citrate

System C

PBS

Acetate

Citrate

Control D

TE

Glycine

PBS

[0050] Table 1

[0051] According to the combination in Table 1, with a single bottle of magnetic beads, we obtained kits A, B, C, and D (control) respectively. In kit A, SPE-C magnetic beads and equilibrium buffer solution 0.25M, pH6.0 citrate, binding buffer 0.15M, pH5.6 phthalic acid, washing buffer 0.35M, pH5.0 lemon salt. In kit B, SPE-C magneti...

Embodiment 2

[0052] Example 2: The process of binding the SPE-C kit of the present invention to polypeptides in serum.

[0053] Take out the above-mentioned kits A-D in the refrigerator at 4°C, and put a tube of SPE-C magnetic bead suspension in it, and manually invert it up and down to mix the magnetic bead suspension evenly. Take 200μl eight-row sample tubes and put them on the orifice plate, add 100μl SPE-C magnetic beads, put the sample tubes on the magnetic bead separator, and let them stand to make the SPE-C magnetic beads adhere to the wall and separate from the suspended liquid. If it is clear, use a sampling gun to absorb the suspended liquid. The tip of the pipette should avoid contact with the SPE-C magnetic beads and avoid absorbing the SPE-C magnetic beads.

[0054] Put the sample tube on the well plate, add 50 μl of SPE-C equilibrium buffer solution, pipette to mix evenly, and mix evenly at 30°C for 18 minutes, put the sample tube on the magnetic bead separator, and let it st...

Embodiment 3

[0056] Example 3: Kit cleaning magnetic beads and polypeptide conjugates

[0057] Place the sample tube on the orifice plate, add 80 μl of SPE-C washing buffer solution, pipette to mix evenly, put it on the orifice plate and let it stand for 5 minutes, put the tube on the magnetic bead separator and let the magnetic beads stick to the wall, and The suspended liquid is separated, the liquid is clear, and the suspended liquid is sucked away with a sampling gun. The tip of the pipette should avoid contact with the magnetic beads to avoid absorbing the magnetic beads. Repeat the (above) step twice to ensure that the suspension is completely aspirated.

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Abstract

The invention provides an SPE-C system suitable for separating protein polypeptide from blood serum, an SPE-C reagent kit thereof and usage of the SPE-C system or the SPE-C reagent kit for separating protein polypeptide from blood serum. The steps are: treating SPE-C magnetic beads in the reagent kit in a balanced way, performing magnetic separation and discarding supernatant; putting the SPE-C magnetic beads in a combination buffer for a specific combination with protein and polypeptide in the blood serum, performing magnetic separation and discarding supernatant; adding a cleaning buffer solution to clean the magnetic beads and remove specific combined impurities; adding elution buffer to elute specific protein and polypeptide in the blood serum, and testing the concentration of the polypeptide; and the elution buffer can be used for mass spectrometry analysis directly or be frozen at 20 DEG C below zero for mass spectrometry analysis within 24 hours. The SPE-C system or SPE-C reagent kit has the capability of capturing more micro molecular and low content proteins, and has the advantages of high flux, low cost, rapidness, simplification, flexible operation, less sample consumption, and wide application to the screening and diagnosis of various malignancy markers.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a SPE-C system suitable for separating proteins and polypeptides in serum and an SPE-C kit for its preparation, more specifically to an SPE-C system or an SPE-C kit for separating Use of protein peptides in serum. Background technique [0002] Proteins are the main executors of various metabolic and regulatory pathways for the survival of biological cells, and the occurrence of diseases may be related to changes in the state of protein modification. Different, the research object of proteome is not a single or a few proteins, it focuses on the study of gene function from a comprehensive, overall, dynamic and quantitative perspective, using the proteome research method from the overall level of cellular protein expression It will be more helpful to understand the biological characteristics of cells through seeing the changes of protein expression in cells. Therefore, the use of proteom...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/18G01N27/64
Inventor 马庆伟李燕张燕刘丽华
Owner BIOYONG TECH
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