Method for concentrating and desalting urine sample

Abstract

The invention relates to a method for concentrating and desalting a urine sample prior to two-dimensional polyacrylamide gel electrophoresis, belonging to the technical field of biological detection. Certain procedures are improved on the basis of the conventional method, and the method for concentrating and desalting the urine sample comprises the following steps of: firstly, performing ultracentrifugation and organic solvent extraction; then, performing dialysis bag desalting and organic solvent extraction. In this way, the salinity is obviously reduced in comparison with the salinity obtained by using the original treatment method. Moreover, the focusing quantity in subsequent isoelectric focusing can reach 60000 vhr, which is beneficial to the successful completion of isoelectric focusing, and the picture forming quality of SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) is improved.

Description

technical field [0001] The patent application of the present invention relates to a method for concentrating and desalting urine samples before two-dimensional polyacrylamide gel electrophoresis, which belongs to the technical field of biological detection. Background technique [0002] Gel electrophoresis, based on the migration of charged proteins in an electric field, is a method commonly used in the biomedical field to separate, identify and purify proteins. The advantage is that proteins are isolated and purified simultaneously, allowing researchers to quickly identify the amount of different proteins in a mixture or the purity of a particular protein preparation. Furthermore, key properties of a protein such as isoelectric point and approximate molecular mass can be determined. Among them, polyacrylamide gel electrophoresis (referred to as PAGE) has a high separation ability and is widely used in the separation and identification of proteins. As a molecular sieve, po...

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Problems solved by technology

Moreover, the voltage often needs to reach more than 5000V during isoelectric focusing. If the salt content is too high, the current will increase during focusing, often exceeding 50μA, which will not only damage the rubber strip but also cause the risk of being broken down by the current.

[0006] There are mainly two types of existing "urine desalination methods": the first method is to use dialysis bags to desalt first, and then extract with organic solvents (including acetone, ethanol, trichloroacetic acid or acetonitrile, etc.), but There are disadvantages that the original sample cannot be effectively concentrated, the desalination effect is poor, and it takes a long time to easily lead to protein degradation in the sample, etc.

The second way is to use an imported desalination kit (or dialysis card) to desalt. Since urine is different from general body fluid samples, the original sample volume is very large, generally about 20-200ml, and the cost of using the second method is very high.

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