Breast cancer cell detection biological reagent and application

A breast cancer cell and biological reagent technology, applied in the field of breast cancer cell detection biological reagents, can solve the problems of high cost, unsuitable large-scale production, cumbersome construction process and the like

Inactive Publication Date: 2018-07-20
刘双萍
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the construction process of the above-mentioned imaging material system is cumbersome and expensive, and is not suitable for large-scale production.

Method used

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  • Breast cancer cell detection biological reagent and application
  • Breast cancer cell detection biological reagent and application
  • Breast cancer cell detection biological reagent and application

Examples

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Embodiment 1

[0021] Example 1 Escherichia coli surface display vector construction

[0022] In this embodiment, the breast cancer cell-targeting bacteria is Escherichia coli BL21 strain, which is transformed into a plasmid containing both the green fluorescent protein (GFP) gene and the breast cancer cell-targeting polypeptide gene, and the C-terminus of the specific polypeptide is connected to the ice nuclei protein The N-terminus of the peptide was displayed on the surface of E. coli, and the green fluorescent protein was used as a marker protein for subsequent detection.

[0023] The specific method is as follows:

[0024] 1. Artificial synthesis of truncated ice nuclei protein

[0025] The polypeptide targeting breast cancer cells is a polypeptide discovered in the previous research of our laboratory. The sequence is SEQ ID NO1, which contains 8 amino acids, and its nucleotide sequence is SEQ ID NO 2.

[0026] SEQ ID NO 1

[0027] Tyr Leu Arg Leu Va l Phe Asn Lys

[0028] SEQ ID 2 ...

Embodiment 2

[0041] Example 2 Breast cancer cell detection biological reagent binding experiment

[0042] Digest skbr3 cells, MCF10A cells, and As578Bst cells respectively, count them, and resuspend them with PBS solution;

[0043] The ratio of immobilized target bacteria and various cells in the breast cancer cell detection biological reagent is 500:1, mix the bacteria and different cells according to the above ratio, and incubate at 4°C for 45min; In the control group expressing the targeted polypeptide, the targeted bacteria were mixed and incubated with various cells according to the same ratio and time. After the above two groups were incubated, they were centrifuged at 5000rpm for 5min, the supernatant was discarded, the precipitate was washed 3 times with PBS solution, the precipitate was resuspended in PBS solution, and analyzed by flow cytometry. The above experiment was repeated 5 times, and the graph was drawn according to the statistical data, and the results were as follows ...

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Abstract

The invention relates to a breast cancer cell detection biological reagent. The breast cancer cell detection biological reagent is formed by breast cancer target bacteria through paraformaldehyde fixation; polypeptides with the sequence of SEQ ID NO.1 are displayed on the surfaces of the breast cancer target bacteria; an expression vector is transferred into a bacterium body to form the breast cancer target bacteria; and a gene encoding the polypeptide, a green fluorescent protein (GFP) gene and an ice nucleation protein N-terminal gene are contained in the expression vector. The breast cancercell detection biological reagent provided by the invention can be used for detection of breast cancer cells.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a biological reagent for detecting breast cancer cells and its application. Background technique [0002] Breast cancer is one of the most common types of tumors in the world. Although some current treatments can help relieve the pain of most cancer patients, breast cancer still poses a great threat to the safety of women's lives, resulting in a decline in the quality of life. Every year, there are more than 1.3 million new cases of breast cancer and 450,000 deaths from the disease worldwide. In breast cancer patients diagnosed at an early stage, the prognosis of surgical treatment is significantly improved compared with that in the middle and late stages, so it is particularly important to improve the accuracy of early diagnosis of breast cancer. [0003] At present, the early diagnosis of breast cancer mainly depends on the detection of clinical medical imaging (B-ultrasound, CT, ...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/569C12N15/70C12N15/65C12N15/62C12N1/21C12R1/19
CPCC07K14/21C07K2319/33C12N15/65C12N15/70G01N33/56966G01N33/57415
Inventor刘双萍
Owner刘双萍