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Applications of isPLA technology based on EML4-ALK antibody in detection of non-small cell lung cancer

A kind of non-small cell lung cancer, technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of difficult to become the clinical standard detection technology of EML4-ALK, limited promotion and use, and the optimal detection method is not determined.

Inactive Publication Date: 2018-07-27
伯仕利生物科技发展(盐城)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although a large number of experimental results have proved the reliability of these three methods, some of their defects have greatly limited their popularization. Although many researchers have compared the effectiveness of RT-PCR, FISH and IHC in the detection of EML4-ALK many times. Specificity and sensitivity, but optimal assay remains undetermined
In addition, researchers have also established a variety of new EML4-ALK detection technologies, but each has certain limitations, and it is difficult to become a clinical standard detection technology for EML4-ALK

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  • Applications of isPLA technology based on EML4-ALK antibody in detection of non-small cell lung cancer
  • Applications of isPLA technology based on EML4-ALK antibody in detection of non-small cell lung cancer
  • Applications of isPLA technology based on EML4-ALK antibody in detection of non-small cell lung cancer

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Embodiment

[0024] The sources of reagents and materials used in the present invention are as follows:

[0025] Materials: Human lung cancer cell lines H2228 and H460 were purchased from ATCC. Human NSCLC tissue samples were provided by Kyungbuk National University Medical Center, South Korea, and all experiments have been approved by the local ethics committee.

[0026] Reagents: ALK-1 antibody was purchased from Cell Signaling Technology; EML4-1 antibody was purchased from Dako; GAPDH antibody was purchased from Superview Biotechnology; ALK Break Apart FISH Probe kit was purchased from Vysis; SP-9002 immunohistochemical staining reagent Box, HRP-labeled goat anti-rabbit IgG, HRP-labeled goat anti-mouse IgG were purchased from Beijing Zhongshan Jinqiao Company; BCA protein quantification kit was purchased from Shanghai Shenggong Bioengineering Co., Ltd.; ECLWestern Blotting kit was purchased from Pierce Company; IllustraTM NAP-5columns Purchased from GE Healthcare; adjacent probe probe arm P...

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Abstract

The invention relates to applications of an ALK(3B12) monoclonal antibody and EML4(1D11) monoclonal antibody combined in-situ proximity ligation assay (isPLA) technology in detection of non-small celllung cancer. According to the present invention, two high-affinity monoclonal antibodies for respectively recognizing EML4 and ALK are prepared by using a monoclonal antibody technology, and by usingthe two high-affinity monoclonal antibodies as the proximity probes, the novel EML4-ALK detection technology (the isPLA technology based on novel EML4 and ALK antibodies) is established; and the technology has advantages of high efficiency, sensitivity, economy, convenience and the like, can be used in clinical diagnosis, and provides the new direction for the detection of tumor markers and treatment targets in the future.

Description

Technical field [0001] The present invention belongs to the field of biotechnology, and relates to the application of isPLA technology based on EML4-ALK antibody in detecting non-small cell lung cancer. Background technique [0002] Lung cancer is a disease caused by the loss of control of cell growth in lung tissue. In the past half-century, the incidence and mortality of lung cancer have increased year by year. It has evolved from a rare disease in the early 20th century to the world's number one cancer killer today. According to the data regularly published by the World Health Organization (WHO), the incidence and mortality of lung cancer are on the rise in countries around the world, especially in developed countries. It has become the most common tumor in many developed countries and ranks first among men with common malignant tumors. It is the second most common malignant tumor in women and women, and it has become the most common cause of death in malignant tumors. In my...

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Application Information

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IPC IPC(8): G01N33/577G01N33/574
CPCG01N33/57423G01N33/57488G01N33/577
Inventor 张炬吴金芸
Owner 伯仕利生物科技发展(盐城)有限公司