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Method for detecting CTCs by multi-antibody immunomagnetic bead enrichment

A technology of immunomagnetic beads and antibodies, applied in the field of immunoassay, can solve the problems of complex testing process, long testing time, high false positive rate, etc., and achieve the effect of short enrichment time, good rigidity, and fast magnetic separation speed

Pending Publication Date: 2019-11-15
济南红枫叶生物科技有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0009] Aiming at the problems of low sensitivity, low detection rate, complicated testing process, long testing time and high false positive rate in the prior art, the present invention uses multiple antibody immunomagnetic beads combined with flow cytometry to detect circulating tumor cells. Greatly improved this set of detection flaws

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  • Method for detecting CTCs by multi-antibody immunomagnetic bead enrichment
  • Method for detecting CTCs by multi-antibody immunomagnetic bead enrichment
  • Method for detecting CTCs by multi-antibody immunomagnetic bead enrichment

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Embodiment 1

[0026] Example 1: 300nm carboxyl magnetic beads were coupled with 1 μmol of HLA-G antibody, 2 μmol of CK8 / 18 antibody, 2 μmol of EPCAM antibody, and 2 μmol of HER-2 antibody, and counted under a fluorescent inverted microscope, and the recovered tumor cells accounted for the pre-enrichment cells The ratio of the total number was used to calculate the tumor cell enrichment efficiency in blood samples by immunomagnetic beads.

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Abstract

The invention relates to a method for detecting CTCs by multi-antibody immunomagnetic bead enrichment. The method comprises specific steps as follows: 1, coupling an anti-EpCAM monoclonal antibody with magnetic beads; 2 preparing a circulating tumor cell sample: adding a proper quantity of immunomagnetic beads to a biological tumor cell sample, performing uniform mixing, then, performing separation by a magnetic separator and PBS cleaning, suspending magnetic bead adsorbed cells in a PBS for cell counting, and detecting enriched cells by a cell counting plate; adding a proper quantity of nanoimmunomagnetic beads to an artificial tumor cell blood sample for magnetic separation, performing PBS cleaning, and then, suspending magnetic bead adsorbed cells in the PBS again; taking the sample, dropwise adding the sample to a glass slide, performing incubation by FICT, detecting enriched cells by a flow cytometer, and identifying effectiveness by tumor cell-fluorescence labeling EPCAM. Counting is performed under a fluorescence inverted microscope, and enrichment and identification of tumor cell line cells mixed with peripheral blood are the same as above. The sensitivity and specificityof circulating tumor cell detection are greatly improved.

Description

technical field [0001] The invention relates to the field of immunodetection, in particular to a method for enriching and detecting CTCs with multiple antibody immunomagnetic beads. Background technique [0002] Detection of circulating tumor cells is helpful for early detection of tumors in patients, monitoring of tumor recurrence or metastasis in patients after surgery, assessment of prognosis and selection of appropriate treatment strategies to help patients treat as soon as possible. In recent years, methods for detecting circulating tumor cells have focused on immune cell therapy, reverse transcription polymerase chain reaction, flow cytometry, immunomagnetic bead enrichment detection, etc., but these methods all have different detection defects: [0003] 1. Immune cell therapy: [0004] The principle of immune cell therapy is to label the specific antibody with a chromogenic agent, and then carry out antigen-antibody reaction in situ on the tissue cells and perform ch...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/6854G01N33/6893G01N33/57407G01N33/57415G01N33/57423G01N33/57442G01N33/57446
Inventor 卢英
Owner 济南红枫叶生物科技有限公司
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