Method for screening and identifying ECH1 autoantibody and application thereof

An autoantibody and identification method technology, applied in the field of ECH1 autoantibody screening and identification, can solve the problem of unsatisfactory tumor markers, and achieve the effects of low cost, high accuracy and simple operation

Active Publication Date: 2017-05-17
郑州臻合生物科技有限公司
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Problems solved by technology

[0005] The purpose of the present invention is to overcome the problem of unsatisfactory existing tumor mark...

Method used

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  • Method for screening and identifying ECH1 autoantibody and application thereof
  • Method for screening and identifying ECH1 autoantibody and application thereof
  • Method for screening and identifying ECH1 autoantibody and application thereof

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Embodiment Construction

[0019] The technical solution of the present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0020] The present invention adopts serum proteomics technology (such as figure 1 ) to screen lung cancer-associated tumor antigens, and use ELISA to detect the level of ECH1 antibody in the serum of lung cancer patients.

[0021] 1) Two-dimensional electrophoresis was performed on the whole protein extract of the lung cancer cell line H1299, and western blotting was performed with the sera of 5 lung cancer patients and 5 normal human sera to screen meaningful tumor autoantigens. After comparison and analysis, they only appeared in the serum of patients The protein spots were further analyzed (such as figure 2 ).

[0022] 2) After the protein spot of interest on the SDS-PAGE gel was cut and digested, the above protein spot was analyzed by LC-MS / MS mass spectrometry method, and it was identified as ECH1 aft...

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Abstract

The invention discloses a method for screening and identifying an ECH1 autoantibody and an application thereof. The method comprises the following steps: adopting a lung cancer cell line H1299 holoprotein extractive for performing two-dimensional electrophoresis; adopting a western blot method for processing the serum of 5 patients with lung cancer and serum of 5 normal people, screening the significant tumor autoantibody, and after comparative analysis, only further analyzing the protein spots appearing in the serum of the patients; cutting and digesting the interesting protein spots on SDS-PAGE gelatin, and then adopting a LG-MS/MS mass spectrometry method for analyzing the protein spots, detecting and identifying as ECH1; and further verifying if the ECH1 autoantibody can be used as a lung cancer diagnosis marker. Through research, the invention finds that the detection for the serum ECH1 autoantibody according to an ELISA method can be used for more accurately identifying the patients with lung cancer from the normal people, identifying the patients with chronic obstructive pulmonary diseases and early diagnosing the lung cancer.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a screening and identifying method of ECH1 autoantibody and its application. Background technique [0002] Lung cancer is a malignant tumor with the highest morbidity and mortality in China and the world. In the past 40 years, the 5-year survival rate of lung cancer has only increased from 12% to 16%. The main reason is that it is diagnosed at an advanced stage. On the contrary, the survival rate of early-diagnosed lung cancer can be increased to 80% after surgery. It can be seen that early detection and early diagnosis have important clinical significance for the treatment and prognosis of lung cancer. Currently widely used detection methods include non-invasive examinations (such as X-ray, CT, mammography, etc.) possibility of widespread use. Finding new lung cancer molecular markers, especially serum molecular markers, so that lung cancer patients can be detected, d...

Claims

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Application Information

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IPC IPC(8): G01N33/574
CPCG01N33/57423G01N33/57488
Inventor 代丽萍王晓李记天张建营
Owner 郑州臻合生物科技有限公司
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