Aptamer-based microfluidic chip capable of capturing cancer cells and preparation thereof as well as separation method of cancer cells

Abstract

The invention discloses an aptamer-based microfluidic chip capable of capturing cancer cells, which is formed by reversibly sealing an upper-layer polymer chip and a lower-layer slide carrier, wherein a cell capturing channel is arranged between the upper-layer polymer chip and the lower-layer slide carrier; the bottom surface of the channel is rough; and avidin is fixed on the bottom surface. The preparation of the microfluidic chip comprises the steps of making a chip die by photoetching, and casting to obtain a polymer chip; processing the slide carrier with hydrofluoric acid and bonding with the polymer chip; and fixing avidin in the cell capturing channel to obtain a microfluidic chip. The microfluidic chip disclosed by the invention can be used for separating cancer cells; and the separation comprises the steps of adding biotin-modified aptamer into a sample, and incubating; and leading the mixed solution after the incubation into the microfluidic chip, wherein the target cancer cells in the mixed solution are combined with the avidin through the biotin to realize separation of the target cancer cells. The microfluidic chip disclosed by the invention has the advantages of high cell capturing efficiency, convenience in operation, strong generality, low cost and the like.

Description

technical field [0001] The invention relates to a cell separation technology, in particular to a microfluidic chip and a separation method which can be used for the separation of cancer cells. Background technique [0002] Cancer, also known as malignant tumor, is a disease caused by the abnormal mechanism of controlling cell growth and proliferation. In addition to growing uncontrollably, cancer cells can also partially invade surrounding normal tissues and even transfer to other parts of the body through the internal circulatory system or lymphatic system, seriously threatening human health. The capture of cancer cells, especially the capture of circulating cancer cells that fall off from the original cancer cell tissue and enter the peripheral blood and tissues of the human body, is of great significance for the early diagnosis of cancer and the evaluation of the therapeutic effect. In the past few decades, people have developed a variety of techniques to capture cancer ...

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Problems solved by technology

Joseph A. Phillips et al. used aptamers as capture probes to effectively separate and enrich cancer cells (Joseph A. Phillips, Ye Xu, Zheng Xia et al Enrichment of cancer cells using aptamers immobilized on a microfluidic channel Anal. Chem. 2009, 81, 1033-1039), but this method cannot solve the influence of laminar flow well due to the simple structure of the chip, and because the nucleic acid aptamer is pre-immobilized on the chip, its contact with cells is small, which makes low The capture efficiency at the concentration is not high

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