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In vivo tumor imaging target molecule and specific probe thereof

A specific and probe technology, applied in the field of molecular imaging, which can solve the problems of limited expression distribution and no biological function of proteins

Inactive Publication Date: 2011-08-24
NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This result shows that although Sp17 is not tumor-specific, due to its limited expression distribution in normal tissues, the expression level in other normal tissues except testis is very small, and the protein has no important biological functions, so it can be used as a target for immunotherapy molecular

Method used

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  • In vivo tumor imaging target molecule and specific probe thereof
  • In vivo tumor imaging target molecule and specific probe thereof
  • In vivo tumor imaging target molecule and specific probe thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055]Example 1 Preparation of anti-sperm protein 17 monoclonal antibody (anti-Sp17 mAb)

[0056] 1. Preparation of anti-Sp17mAb: inject liquid paraffin (0.5ml / mouse) into the peritoneal cavity of mice, and inject hybridoma cells (0.5ml / mouse) with the preservation number CGMCC No.1434 into the peritoneal cavity of mice 14 days later. When the abdomen of the mice was obviously enlarged, the ascites of the mice was extracted with a No. 8 needle, and the ascites was extracted in the same way every 4 days until the mice died. Centrifuge the ascites at 3000rpm for 10 minutes, discard the upper layer of fat and insoluble matter and various cells and insoluble matter in the lower layer, absorb the light yellow supernatant, add an equal volume of glycerol (containing 20g / L Na 2 HPO 4 ), and stored in a -20°C refrigerator after aliquoting.

[0057] 2. Purification of anti-Sp17mAb

[0058] 1) Ammonium sulfate salting out: Add 500g of ammonium sulfate into 500ml of distilled water, h...

Embodiment 2

[0062] Example 2 Establishment of tumor animal models

[0063] BALB / C nude mice (male or female, 4-6 weeks old, 20-25 g). Ovarian cancer cell line HO8910 or liver cancer cell line SMMC-7721 overexpressing Sp17 were cultured in RPMI1640 medium. When the cells cover more than 80% of the bottom of the bottle, digest the cells with 0.25% trypsin-0.02% EDTA for 1 min, stop the reaction with RPMI1640 culture medium containing 10% calf serum, blow and beat to disperse the cells, centrifuge at 1000rpm for 5 min, discard the supernatant, and use PBS to adjust the number of cells, 2 × 10 6 / 0.2ml, the cells were inoculated subcutaneously in the shoulders or buttocks of 20-25g nude mice. After about 10-20 days, the tumors gradually grew.

Embodiment 3

[0064] Example 3 Preparation of Immunomagnetic Nanoprobes (IMNPs)

[0065] Using chemical coupling agent EDC / NHS (1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide / N-hydroxysuccinimide N-hydroxysuccinimide), the two Magnetic nanoparticles with different surface modifications (nanoscale γ-Fe 2 o 3 Particles, prepared by the School of Biological Science and Medical Engineering of Southeast University by co-precipitation method) were coupled with anti-Sp17mAb to prepare immunomagnetic nanoprobes (anti-Sp17IMNPs, hereinafter referred to as IMNPs), and compared the targeting effect of the two on tumors.

[0066] 1. Anti-Sp17-Silane@MNPs probe preparation: Take 0.2 mg of silane-modified magnetic nanoparticles (Silane@MNPs), add 100 μg of anti-Sp17 mAb activated by EDC / NHS, mix well, and shake at room temperature for 2 hours. Then the mixture was magnetically separated and washed with PBS buffer three times to remove free antibodies, and finally the precipitate was suspended in PBS (0...

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Abstract

The invention belongs to the field of molecular imaging, and discloses an in vivo tumor imaging target molecule and a specific probe thereof. The target molecule is sperm protein 17 (Sp17) which is expressed abnormally in a tumor cell; the specific probe comprises targeting agent and tracer; and the targeting agent is monoclonal antibody capable of being combined with Sp17 specific, and the tracer is fluorescent dye or radioactive nuclide. The probe can be combined with the specific of an in vivo tumor, has good imaging effect and good stability, and has good application prospect in preparingtumor molecular imaging diagnostic reagent.

Description

technical field [0001] The invention belongs to the field of molecular imaging, and relates to a living tumor imaging target molecule and a specific probe thereof. Background technique [0002] Molecular imaging is a science that uses imaging methods to display specific molecules at the organ level, tissue level, and cell level, reflects the changes at the molecular level in the living state, and conducts qualitative and quantitative research on its biological behavior in imaging. Molecular imaging is the result of the unprecedented development of molecular biology and cell biology technology in recent years, as well as the application of transgenic animal models, emerging and efficient imaging probes and drugs. It is a combination of molecular biology technology and modern medical imaging. A newly emerging fringe discipline. Classical imaging diagnosis (CT, MRI, etc.) mainly shows the final effect of some molecular changes, and diseases with anatomical changes; while molec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K51/08A61K103/10
CPCC12Q1/6886C12Q2600/158
Inventor 李芳秋张士新刘群
Owner NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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