Unlock instant, AI-driven research and patent intelligence for your innovation.

Molecular probe related to disease and method for producing the same

A molecular and disease technology, applied in the field of molecular probes and their preparation, can solve the problems of few aptamer molecules, few pure biomolecules, low screening efficiency, etc., and achieve a wide range of applications, high stability, and specific binding. strong effect

Inactive Publication Date: 2012-06-20
谭蔚泓
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few nucleic acid aptamer molecules currently available for detection, which cannot be widely used by researchers like a wide variety of antibodies.
There are many reasons for this situation, one of which is that the current screening efficiency is not high, and there are not many aptamers screened out; another reason is that nucleic acid aptamers need to be screened with pure target molecules, which are currently available. Very few pure disease-related biomolecules are available
Therefore, it is difficult to obtain a large number of nucleic acid aptamers for diagnosis by conventional methods, and a new method for screening disease-related nucleic acid aptamers is needed to prepare disease-related molecular probes

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Molecular probe related to disease and method for producing the same
  • Molecular probe related to disease and method for producing the same
  • Molecular probe related to disease and method for producing the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1. Preparation of molecular probes related to leukemia

[0042] 1. Screening of nucleic acid aptamers related to leukemia

[0043] (1) Synthesis of random nucleic acid library

[0044] According to the literature (Jayasena S D. Aptamers: an emerging class of molecules thatrival antibodies in diagnostics. Clin. Chem. 1999, 45: 1628-1650), the following random single-stranded DNA library was designed and synthesized: 5'-ATA CCA GCT TAT TCA ATT-52-nt-AGA TAGTAA GTG CAA TCT-3'.

[0045] (2) Screening of nucleic acid aptamers related to leukemia

[0046] Binding buffer: containing 4.5g / L glucose, 5mM MgCl 2 , 1mM CaCl 2 , 2.67mM KCl, 1.47mM potassium dihydrogen phosphate, 137.93mM sodium chloride, 8.06mM disodium hydrogen phosphate, 0.1mg / mL tRNA (R8759 Sigma, Ribonucleic acid, transfer from baker's yeast (S.cerevisiae)) and 1mg / mL bovine serum albumin (Fisher), pH 7.4.

[0047] CCRF-CEM cells (human T lymphocyte acute lymphoblastic leukemia) (CCL-119) and Ram...

Embodiment 2

[0084] Example 2. Molecular probes are used in the detection of leukemia cells

[0085] The three probes ppA, pB and pC obtained in Example 1 were used to detect leukemia cells respectively. Take 1μM molecular probe and incubate for 30 minutes with the isolated leukemia cells and the clinical bone marrow samples of insiders, centrifuge, wash the cells twice, and detect the cells with a flow cytometer.

[0086] The experiment was repeated 3 times and the results were consistent.

[0087] The detection results of the ppA molecular probe are as follows: image 3 Shown (wherein 1 represents the detection result of the leukemia cell CCRF-CEM, and the rest are the detection results of the normal cells (control cells) in the bone marrow). The pB and pC molecular probes have similar effects. It shows that the molecular probe of the present invention can effectively identify leukemia cells and specifically bind to it, and the molecular probe of the present invention can be used for ...

Embodiment 3

[0088] Example 3. Molecular probes are used in disease cell typing research

[0089] The molecular probe sequence related to lymphoma was prepared according to the method described in Example 1. The difference in the method was that lymphoma cells were taken as target disease cells, and 2 of them were labeled with fluorescein as molecules related to lymphoma probe.

[0090] Take 1 μM molecular probes related to lymphoma and the above-mentioned ppA, pB, pC molecular probes, incubate with leukemia cells and lymphoma cells for 30 minutes, centrifuge, wash cells twice, and use flow cytometry to detect different molecules Phenotypic characterization of probe binding to different cells.

[0091] The experiment was repeated 3 times and the results were consistent. The results show that the binding profiles of lymphoma cells and different molecular probes are similar, and the binding profiles of leukemia cells and different molecular probes are similar; it shows that the probes obta...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a molecular probe related to disease and a method for producing the same. The method for screening aptamer related to the disease comprises the steps as follows: a nucleic acid library is screened by target disease cell in vitro to obtain aptamer related to the target disease; the nucleic acid library is library of single stranded random nucleotide sequence. The method of the invention can screen the aptamer related to the disease under the condition of not knowing the molecular transformation of the disease cell surface related to the disease beforehand, and can obtain multiple aptamers related to the disease. So, the method of the invention is widely used and has wide application range. Proved by experiment, the method of the invention can at once screen 3 to 20 aptamers for identifying the different molecules of the cell surface; after modification, the specificity bonding force between the aptamer and the disease cell is stronger and the stability in the biological environment is higher.

Description

technical field [0001] The invention relates to a molecular probe related to a disease and a preparation method thereof. Background technique [0002] The occurrence and development of diseases lead to changes in the types and quantities of molecules in cells. Therefore, detecting and tracking these changes at the molecular level is essential to understand the mechanism of disease, improve diagnostic accuracy, advance the diagnostic window period, formulate reasonable treatment plans, and monitor treatment. A very effective way of effect. However, due to the lack of effective disease-specific molecular probes, it is still a huge challenge to understand and diagnose diseases at the molecular level (Espina V, Geho D, Mehta AI, Petricoin EF 3rd, Liotta LA, & Rosenblatt KP. Pathology of the future: molecular profiling for targeted therapy. Cancer Invest 2005; 23:36-46). The diagnosis of most diseases (such as tumors) still mainly relies on the traditional tissue and cell morph...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C40B30/04C12P19/34C12N15/10C12N15/11
Inventor 谭蔚泓上官棣华方晓红王柯敏杨朝勇
Owner 谭蔚泓