Erythropoietin detection method based on magnetic bead as carrier

Abstract

The invention relates to the technical field of erythropoietin, and discloses an erythropoietin detection method based on a magnetic bead as a carrier, which comprises the following steps of: coupling a capture antibody (anti-EPO antibody) on the magnetic bead, capturing EPO in serum by the capture antibody coupled on the magnetic bead, and then adding a biotinylated detection antibody, a'capture antibody-EPO-detection antibody 'immune complex taking magnetic beads as a carrier is formed, then streptavidin coupled enzyme is added to be combined with biotin labeled on the detection antibody, and the immune complex is labeled. The established immunoassay method has ultrahigh sensitivity (19pg / mL) which is improved by 13 times compared with the traditional immunoassay method, the sample dosage (15mu L) is reduced by 13 times compared with the traditional immunoassay method, the sample detection experiment period is shortened to 34min which is far shorter than the traditional immunoassay method (6-8h), the change rule of endogenous EPO in serum can be detected more timely by using the method, and the method has the advantages of high sensitivity and high sensitivity. And a dynamic change process of absorption, distribution, metabolism, excretion and the like of exogenous EPO in a body is supported.

Description

technical field [0001] The invention relates to the technical field of erythropoiesis, in particular to an erythropoietin detection method based on magnetic beads as a carrier. Background technique [0002] Human erythropoietin (EPO) is a protein that promotes erythroid progenitor cells (CD34 + Hematopoietic stem cells) is a glycoprotein hormone that proliferates, differentiates, and matures erythrocytes. It is secreted by the kidney and liver in fetuses, and is mainly produced by the kidneys in adults. It is secreted into the blood circulation and acts on erythroid progenitor cells in the bone marrow. EPO binds to specific receptors on target cells, such as bone marrow cells, spleen cells, and fetal liver cells, thereby promoting the proliferation and differentiation of red blood cell precursor cells into mature red blood cells, increasing the amount of red blood cells in the blood circulation from the bone marrow, and maintaining red blood cells in the blood. amount in th...

AI Technical Summary

Problems solved by technology

[0003] Due to the existence of endogenous EPO in serum, after the body is given exogenous EPO, it will cause great interference to the detection of exogenous EPO in serum. At present, the detection methods for EPO are mainly divided into two categories, one of which is Radioimmunoassay, and the other is immunoassay; radioimmunoassay requires a laboratory with special qualifications to carry out the test because there are radioactive substances, and the experimental process will generate a large amount of radioactive waste, which requires a large amount of material resources and financial resources to control and dispose of radioactive waste, in addition, the energy spent on operator qualifications and protection is also relatively large

The traditional immunoassay (ELISA) is interfered by endogenous EPO, the sensitivity of the method is about 250pg / mL, and the sample volume is large. Each detection requires about 200μL blood sample, and the experiment time is more than 6 hours. Due to the limitation of sensitivity, Immunoassays cannot be used to detect endogenous EPO expression and serum drug concentrations after exogenous EPO administration

It has caused great obstacles to the detection of EPO in vivo and the description of the dynamic change process of exogenous EPO in vivo

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