Negative-phase enrichment method and kit for rare cells in blood based on microfluidics and immunomagnetic separation

Abstract

The invention relates to a negative-phase enrichment method and kit for rare cells in blood based on microfluidics and immunomagnetic separation. The negative-phase enrichment method comprises the following steps: (1) subjecting a blood sample to centrifugation with a pre-treatment fluid so as to remove a part of impurities in blood; (2) subjecting the sample treated in the step (1) to lysis with a lysis solution, removing erythrocytes in blood and collecting residual cells; (3) adding magnetic particle-bonded leucocytes connected with corresponding antibodies into the residual cells and carrying out immunoreactions at room temperature; (4) subjecting the sample treated in the step (3) to separation with a separating medium to remove unbonded magnetic particles; and (5) carrying out separation by using an immunomagnetic particle capturing instrument and micro-fluidics and removing leucocytes so as to an enriched sample. The invention also provides the corresponding kit. According to the invention, erythrocytes are subjected lysis; and a plurality of technologies like immunomagnetic separation, density gradient centrifugation and micro-fluidics are optimally combined to rapidly remove leucocytes, and the removal rate of leucocytes reaches 99% or above.

Description

technical field [0001] The invention belongs to the field of cell enrichment, in particular to a negative phase enrichment method and kit for rare cells in blood based on microfluidics and immunomagnetic separation. Background technique [0002] CTCs (Circulating Tmor Cells) are defined as tumor cells that escape from the primary tumor or metastases of solid tumors and enter the peripheral blood circulation spontaneously or due to diagnostic and therapeutic procedures. Since 2003, Professor Cristofanilli of the University of Texas Anderson Cancer Center and his team have first confirmed that the number of CTCs before treatment can be used as the progression-free survival rate (PFS) of patients with metastatic breast cancer in the literature published in the New England Journal of Medicine. After being an independent predictor of overall survival (OS), the detection of CTCs has been gradually promoted and applied in scientific research and clinical fields. In the detection o...

AI Technical Summary

Problems solved by technology

The current routine leukocyte removal is mainly through centrifugation, immunization, etc., and there are also various methods such as affinity chromatography and magnetic particles for removal using the principle of immunity. However, the immune antibody used is usually only a single anti-CD-45 antibody. Although anti-CD-45 CD-45 antibody is the antibody with the highest binding rate to leukocytes among the existing CD series antibodies, but the results still have room for improvement

Images