Specific antibody delivery platform and preparation method and application thereof

A specific and species-specific technology, applied in the field of molecular biology, can solve problems such as immature regulatory means, and achieve the effect of promoting NK cell migration, improving effect and wide application prospect.

Pending Publication Date: 2021-06-22
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, the regulation of cell migration in vivo is still immature, and there are still many unknowns to be explored

Method used

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  • Specific antibody delivery platform and preparation method and application thereof
  • Specific antibody delivery platform and preparation method and application thereof
  • Specific antibody delivery platform and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Preparation of MNP@NK1.1

[0048] Experimental Materials:

[0049] Anti-mouse NK1.1 antibody was purchased from Bio Cell, USA; MNP was purchased from Xi'an Ruixi Biotechnology Co., Ltd.; EDCI was purchased from Tokyo Chemical Industry Co., Ltd. (TCI); NHS was purchased from Aladdin, Shanghai.

[0050] The preparation process is as figure 1 As shown, first combine MNP with an activator for esterification reaction, and activate some carboxyl groups on the surface into active esters; then react the obtained MNP with active esters with amino groups on antibodies that can specifically recognize NK cells to obtain Magnetic nanoparticles that recognize and bind NK cells.

[0051] Performance characterization and optimization:

[0052] 1.1 Optimization of activator concentration:

[0053] The carboxyl group on the surface of the particle can undergo esterification reaction under the action of the activator to generate an active ester, which further interacts with ...

Embodiment 2

[0071] Example 2 The specific binding of MNP@NK1.1 particles to NK cells

[0072]Experimental materials: the mouse B16-F10 melanoma cell line was obtained from the American Standard Biological Collection (ATCC). SPF grade female C57BL / 6 mice, 5-6 weeks old, were purchased from Hunan Slack Jingda Experimental Animal Co., Ltd. The mice were kept in the Experimental Animal Center of South China University of Technology, and the animal experiment procedures followed the relevant provisions of the South China University of Technology's Regulations on the Management of Experimental Animals.

[0073] Anti-mouse NK1.1 antibody was purchased from Bio Cell, USA.

[0074] 2.1 Binding of MNP@NK1.1 particles to NK cells

[0075] To evaluate the ability of magnetic nanoparticles MNP@NK1.1 to bind NK cells, we first explored the ability of MNP@NK1.1 particles to interact with NK cells. Firstly, the NK cells in the spleen of C57B / 6 mice were sorted out by the NK cell magnetic bead sorting ...

Embodiment 3

[0083] Example 3 MNP@NK1.1 particles promote NK cell migration under the action of magnetic field

[0084] In order to verify whether MNP@NK1.1 particles can promote the migration of NK cells under the action of an external magnetic field, we co-incubated NK cells with MNP@NK1.1 particles in a culture dish. Cell density is 1 x 10 6 cells / mL, the particle concentration is 25 μg / mL, and the incubation condition is 30 min in an incubator at 37°C.

[0085] Test results such as Figure 9 As shown, NK cells gathered at the edge of the culture dish under the action of an external magnetic field, indicating that the particles have the ability to promote the migration of NK cells.

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Abstract

The invention relates to a specific antibody delivery platform and a preparation method and application thereof. The preparation method of the specific antibody delivery platform comprises the steps that NK cells serve as target spots, the surfaces of ferroferric oxide nanoparticles are modified, and the ferroferric oxide nanoparticles and the antibody for specifically recognizing the NK cells are combined through chemical bonding. According to the specific delivery platform, the antibody and the particles are combined more stably in a chemical bonding mode, the NK cells can be specifically recognized and combined, and migration of the NK cells can be further promoted under the action of a magnetic field, so that enrichment of the NK cells at tumor sites can be rapidly and efficiently achieved, and the immunotherapy effect can be obviously improved. Therefore, the platform has a wide application prospect in medical detection, blood screening and targeted drug preparation.

Description

technical field [0001] The present invention relates to the field of molecular biology, in particular to a specific antibody delivery platform and its preparation method and application. Background technique [0002] NK cells are an important part of the innate immune system. NK cells have a variety of immune functions, which can not only directly kill cells but also secrete pro-inflammatory factors to enhance the body's immune response. NK cells have the ability to directly kill tumor cells, and anti-tumor immunotherapy based on NK cells has developed rapidly in the 21st century. Although these therapies have not achieved great clinical success like adoptive T cell therapy, the results of preclinical and clinical trials have highlighted the great potential of NK cell-based treatment of tumors. In the process of NK cells functioning, NK cells need to be enriched in the tumor site under the action of pro-inflammation, and then recognize the tumor through the "missing self" ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K17/00C07K16/28A61K47/69A61K47/68A61K41/00A61P35/00B82Y5/00G01N33/543G01N33/574G01N33/68
CPCA61K41/00A61K47/68A61K47/6923A61P35/00B82Y5/00C07K16/2803C07K16/2842C07K16/2851C07K17/00G01N33/54346G01N33/574G01N33/6854G01N2800/52
Inventor 王均张鹏飞沈松叶倩妮
Owner SOUTH CHINA UNIV OF TECH
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