Carcino-embryonic antigen monoclonal antibody, chip containing same and application

A carcinoembryonic antigen and antibody chip technology is applied in the field of bioengineering to achieve accurate qualitative detection, convenient operation and strong specificity

Inactive Publication Date: 2012-12-26
李彬
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But smoking, pregnancy, cardiovascular disease, diabetes, non-specific colitis and other diseases, 15% to 53% of patients wi

Method used

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  • Carcino-embryonic antigen monoclonal antibody, chip containing same and application
  • Carcino-embryonic antigen monoclonal antibody, chip containing same and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: Preparation of the monoclonal antibody of the present invention

[0045] The preparation method of antibody comprises the following process steps:

[0046] (1) Cell fusion: Sp2 / 0 myeloma cells with high activity were mixed with splenocyte suspensions sensitized with self-designed and synthesized carcinoembryonic antigen polypeptide (sequence TQQAT PGPAYSGREI) at a ratio of 1:10-100, and added Polyethylene glycol (produced by Sigma, USA) makes the cells fused with each other, and the culture medium is added dropwise to the mixed cell suspension of the two cells, and the cells are cultured with HAT selective medium (produced by Haiclone, USA);

[0047] (2) Screen hybridoma cells: when the cells to be fused are cultured to the 5th to 10th day, absorb the culture supernatant of the clonal cell clusters in the wells of the 96-well culture plate, and use the enzyme-linked immunosorbent assay method to detect the antibody content, and limit dilution Three times ...

Embodiment 2

[0068] Embodiment 2: the preparation method of antibody chip

[0069] Increase following processing steps on the basis of embodiment 1:

[0070] Step (5) Label complement C3 antibody with CY3 fluorescein.

[0071] Step (6) Spot 96 spots on one chip, two spots for each monoclonal antibody, and the content of each spot is 0.01-1ng / ml.

Embodiment 3

[0072] Embodiment 3: Detection of various indicators of the monoclonal antibody described in the present invention

[0073] The monoclonal antibody secreted by the hybridoma cell line screened in Example 1 was subjected to sequence analysis, and its heavy chain variable region was as follows:

[0074] VKLQESGPGL VAPSQSLSMS CTVSGFSLSS YGVHWVRQPPGKGLEWLGVI WAGGTTNYNS ALMSRLSISK DNSKSQVLLKMNSLQTDDTA MYYCATTTMI TLMDYWGQGT TVTVSSA (shown in SEQ ID No. 1)

[0075] Its light chain variable region is as follows:

[0076] DVQLNQAKSS LSASLGDRVT ISCRASQDIS NYLNWYQQKP DGTVKLLIYY TSRLHSGVPP RFSGSGSGTD YSLTISNLEQ DIATYFCQQGNTVPWTFGG GTKLEIS (shown in SEQ ID No. 2)

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Abstract

The invention relates to the technical field of bioengineering and discloses a monoclonal antibody of carcino-embryonic antigen and a chip containing the antibody. A heavy chain variable region amino acid sequence of the monoclonal antibody is shown as SEQ ID NO.1, and a light chain variable region amino acid sequence is shown as SEQ ID NO.2. A reaction result of the antigen antibody is convenient for quality control due to the homogeneity and the biological activity unicity of the monoclonal antibody, which is beneficial to standardization and normalization. The chip containing the antibody can be directly applied to medical and scientific research institutions to diagnosing and preventing various tumor diseases, and the technical problems of time and labor consumption, poor result repeatability and the like of the traditional technology are solved, so that the invention has extremely high application values in the field of clinical diagnosis and wide application prospect.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a carcinoembryonic antigen monoclonal antibody, a chip containing the antibody and its application. Background technique [0002] Carcinoembryonic antigen (CEA) is an acidic protein, originally found in colon cancer and fetal intestinal tissue, hence the name carcinoembryonic antigen. The content of CEA in the normal mucosa adjacent to the cancer was little or negative. The positive rate of CEA in gastric cancer was 85.58%. Among them, mucinous adenocarcinoma and signet ring cell carcinoma (mucinous cell carcinoma) were 100%. Under the electron microscope, it can be seen that CEA is distributed in the membrane of cancer cells, and in the organelles of protein synthesis and transport in the cytoplasm of cancer cells (such as nuclear membrane, endoplasmic reticulum, Golgi apparatus and its secretory vesicles). Elevated CEA is common in colorectal cancer, pancreatic cancer...

Claims

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Application Information

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IPC IPC(8): C07K16/30G01N33/68G01N33/577C40B40/10
Inventor 李彬
Owner 李彬
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