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Application of Citric Acid Alcohol Method in Removing Albumin from Serum

A technology of albumin and citric acid, applied in the field of serum albumin removal, can solve the problems of limited albumin removal and unsatisfactory albumin removal effect, and achieve the effects of safe and low toxicity of reagents and convenient operation.

Inactive Publication Date: 2015-07-29
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the removal effect of the above methods on albumin is not very satisfactory. The removal of albumin by DAC method is limited. Although more albumin is removed by TCA method, some other trace proteins are also removed at the same time.

Method used

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  • Application of Citric Acid Alcohol Method in Removing Albumin from Serum
  • Application of Citric Acid Alcohol Method in Removing Albumin from Serum
  • Application of Citric Acid Alcohol Method in Removing Albumin from Serum

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0028] Experimental example 1 citric acid ethanol method removes serum albumin

[0029] figure 1 It is the chemical structural formula of citric acid, and the protein extraction process is carried out as follows:

[0030] 1) Serum centrifugation (4°C, 15000g, 15min) to remove lipids in the blood;

[0031] 2) absorb 20 μl of serum after degreasing and add 60 μl of extract, wherein the extract is an anhydrous ethanol solution containing 3% citric acid by weight and volume, and vortex to mix;

[0032] 3) The serum extract was allowed to stand at -20°C for 30 minutes;

[0033] 4) The above serum extract was centrifuged (4°C, 15000g, 15min), the precipitate was retained, and the supernatant was transferred to a new EP tube;

[0034] 5) The precipitate in step 4 was evaporated on a rotary evaporator for 5 minutes to remove residual organic solvent, and the precipitate was stored.

[0035] 6) The supernatant in step 4 was evaporated on a rotary evaporator for 25 minutes, the orga...

experiment example 2

[0036] Influence of Experimental Example 2 Different Organic Solvents on Albumin Removal Effect

[0037] Albumin was removed with different organic solvents according to the method of Experimental Example 1. The organic solvents used were methanol containing 3% citric acid, absolute ethanol, acetone, and propanol. The results showed that the application of absolute ethanol to extract serum protein can achieve good results. The result is as Figure 4 Shown: M is the standard protein SDS6H2, band 1 is the original serum, and bands 2-5 are the precipitated proteins after the treatment of the extract (the solvents of the extract are respectively methanol, absolute ethanol, acetone, and propanol) electrophoresis bands; Bands 6-9 are electrophoresis bands of supernatant albumin after treatment with extract solution corresponding to bands 2-5.

experiment example 3

[0038] Experimental Example 3 Effects of Different Acids on Albumin Removal Effect

[0039] According to the method of Experimental Example 1, different acids were used to remove albumin. The acids used were phosphoric acid, acetic acid, citric acid, and TCA, and the results showed that citric acid had the best extraction effect. Such as Figure 5 Shown: band M is the standard protein SDS6H2, band 1 is the original serum, bands 2-5 are the electrophoresis bands of the precipitated protein after treatment with the extract solution, and bands 6-9 are the electrophoresis bands corresponding to 2-5 after treatment with the extract solution The supernatant albumin electrophoresis bands.

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Abstract

The invention discloses application of a citric acid-ethanol method in removal of albumin in serum, and relates to a method for removing high-abundant protein from serum, in particular to a new method for removing albumin from serum. The method comprises the following steps of: 1) centrifuging the serum to remove lipid; 2) adding a serum protein extracting solution, mixing uniformly, putting the mixture into a refrigerator of 20 DEG C below zero, and standing for 30 minutes; 3) centrifuging the sample extracting solution; and 4) respectively collecting the centrifuged sediment and the supernate. The method for removing the albumin from the serum is easy to operate, low in price and short in required time.

Description

technical field [0001] The invention relates to a method for removing high-abundance proteins in serum, in particular to a new method for removing serum albumin. Background technique [0002] As a medical discipline, hematology has made great progress with the development of molecular biology and related technologies, and has great application prospects in the early diagnosis and diagnosis of diseases. In clinical disease diagnosis, changes in blood components have a very important relationship with the occurrence and development of diseases. In recent years, the use of proteomics technology to explore blood has become a research hotspot. [0003] Due to the extremely complex components of plasma, effective separation and analysis of them has become the key to blood proteomics research. However, some high-abundance proteins often mask the detection of low-abundance proteins. It has been reported that about 22 high-abundance proteins account for 98% of the total plasma prot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/34
Inventor 丛维涛金利泰张鹏辉洪国赢林丽朱忠欣李校堃
Owner WENZHOU MEDICAL UNIV
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