Novel CTC (circulating tumor cell) enrichment technology and preparation method of kit

A technology of tumor cells and kits, applied in the medical field, can solve the problems of limited selection ability, low quantity, weak signal, etc., and achieve the effect of convenient, fast and accurate detection

Inactive Publication Date: 2014-04-16
CHANGSHA YINGRUN BIOTECH
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Problems solved by technology

The current enrichment of circulating tumor cells mainly adopts the method of forward sorting, which directly sorts tumor cells of mononuclear cells in peripheral blood by antibodies and magnetic beads or flow cytometry. This method has disadvantages: 1. Single cells in peripheral blood The number of ...

Method used

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  • Novel CTC (circulating tumor cell) enrichment technology and preparation method of kit
  • Novel CTC (circulating tumor cell) enrichment technology and preparation method of kit
  • Novel CTC (circulating tumor cell) enrichment technology and preparation method of kit

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Embodiment 1

[0029] A new circulating tumor cell enrichment technique, comprising the following steps:

[0030] (1) Arrange and combine all possible CD45 isoform protein sequences, and prepare various corresponding antigens through careful analysis, so as to obtain various antibodies;

[0031] (2) Screen out antibodies that can recognize all CD45 isoforms by making a variety of antibodies, couple them to magnetic beads, and then capture all white blood cells from the blood, where the antigen sequence includes the full length of the extracellular region , the full length of the extracellular region spliced ​​by different exons, aiming to screen out an antibody that can recognize their common segment, the reason why the orange protein region is not used alone is because we want to more realistically simulate the protein of the outer part of the cell membrane Conformation, directly screen our antibodies with a variety of CD45 isoform antigens, so that the obtained antibodies can directly reco...

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Abstract

The invention discloses a novel CTC (circulating tumor cell) enrichment technology and a preparation method of a kit. The technology comprises the steps as follows: through permutation and combination of all possible CD45 isomer protein sequence information, designing antigens, thereby producing multiple antibodies, screening out one antibody capable of recognizing all CD45 isomers, coupling the antibody onto a magnetic bead, and further capturing all leukocytes from blood; collecting 10 ml of peripheral venous blood; removing red blood cells through lysis by an osmosis lysis buffer; mixing a designed specific antibody, the magnetic bead and the rest cell sap containing no red blood cells to separate leukocytes; and obtaining a cell population containing CTCs with higher concentration. According to the technology, a negative collection technology is adopted, so that the proportion of the CTCs is increased substantially, and the rapid and accurate detection is facilitated; meanwhile, a negative screening means is adopted, so that the CTCs are reserved without loss, and the fact that tumor diseases cannot be detected and the treatment for patients is delayed due to the loss of certain CTCs is prevented.

Description

technical field [0001] The invention relates to the field of medicine, in particular to a new circulating tumor cell enrichment technology and a preparation method of a kit. Background technique [0002] Circulating tumor cells (CTCs) refer to tumor cells released from solid tumors or metastases into the peripheral blood circulation spontaneously or due to diagnostic and therapeutic procedures, and are an important cause of postoperative recurrence and distant metastasis in patients with malignant tumors. In recent years, with the continuous improvement of detection technology, CTCs detection, as a new type of non-invasive diagnostic tool, has become the focus of clinical research for its application value in early detection of postoperative recurrence and distant metastasis, evaluation of curative effect and prognosis, etc. hotspot. [0003] Molecular biology and clinical studies have shown that tumor invasion and micrometastasis are likely to appear in the early stages of...

Claims

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Application Information

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IPC IPC(8): C12N5/09
Inventor 张万明易银沙朱海强李娜刘彩云袁炳秋吴小宁
Owner CHANGSHA YINGRUN BIOTECH
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