Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Fluorescent dye labeling aptamer for immune globulin E with sensitive fluorescence anisotropy response

A nucleic acid aptamer and immunoglobulin technology, applied in the field of analysis and detection, can solve the problems of no obvious signal change, influence sensitivity, and insignificant fluorescence polarization, and achieve the effect of significant signal change, large signal change range and good affinity.

Active Publication Date: 2018-04-03
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the end-labeled aptamer such as fluorescein or tetramethylrhodamine binds to the protein, the change in fluorescence polarization (fluorescence anisotropy) caused is sometimes not significant, which affects the analysis of fluorescence polarization (fluorescence anisotropy) sensitivity, even resulting in no appreciable signal change

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
  • Fluorescent dye labeling aptamer for immune globulin E with sensitive fluorescence anisotropy response
  • Fluorescent dye labeling aptamer for immune globulin E with sensitive fluorescence anisotropy response
  • Fluorescent dye labeling aptamer for immune globulin E with sensitive fluorescence anisotropy response

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: Detection of IgE using IgE37-T9-FAM fluorescence anisotropy (fluorescence polarization)

[0066] 1. Mix IgE37-T9-FAM, IgE and binding buffer to obtain a reaction system. The final concentration of IgE37-T9-FAM in the reaction system was 10 nM. The final concentrations of IgE in the reaction system were 0nM, 0.01nM, 0.02nM, 0.05nM, 0.1nM, 0.2nM, 0.5nM, 1nM, 2nM, 5nM, 10nM, 20nM, 50nM, 100nM and 200nM, respectively.

[0067] The above-mentioned IgE37-T9-FAM is an aptamer obtained by FAM-labeling the ninth nucleotide T of the nucleic acid aptamer IgE37.

[0068] 2. Incubate the reaction system in an ice box (4°C) for 20 minutes.

[0069] 3. The fluorescence anisotropy value (r) of IgE37-T9-FAM was measured at 25°C using a fluorometer.

[0070] The result is as figure 1 shown. figure 1 It was shown that the fluorescence anisotropy of IgE37-T9-FAM increased gradually with the increase of IgE concentration. At high concentration of IgE, the fluorescence anisot...

Embodiment 2

[0071] Example 2: Detection of IgE using IgE37-T10-FAM fluorescence anisotropy (fluorescence polarization)

[0072] 1. Mix IgE37-T10-FAM, IgE and binding buffer to obtain a reaction system. The final concentration of IgE37-T10-FAM in the reaction system was 10 nM. The final concentrations of IgE in the reaction system were 0nM, 0.01nM, 0.02nM, 0.05nM, 0.1nM, 0.2nM, 0.5nM, 1nM, 2nM, 5nM, 10nM, 20nM, 50nM, 100nM and 200nM, respectively.

[0073] The above-mentioned IgE37-T10-FAM is an aptamer obtained by labeling the 10th nucleotide T of the nucleic acid aptamer IgE37 with FAM.

[0074] 2. Incubate the reaction system in an ice box (4°C) for 20 minutes.

[0075] 3. The fluorescence anisotropy value (r) of IgE37-T10-FAM was measured at 25°C with a fluorometer.

[0076] The result is as figure 1 shown. figure 1 It is shown that the fluorescence anisotropy of IgE37-T10-FAM increases gradually with the increase of IgE concentration. At high concentration of IgE, the fluorescenc...

Embodiment 3

[0077] Example 3: Detection of IgE using IgE37-T11-FAM fluorescence anisotropy (fluorescence polarization)

[0078] 1. Mix IgE37-T11-FAM, IgE and binding buffer to obtain a reaction system. The final concentration of IgE37-T11-FAM in the reaction system was 10 nM. The final concentrations of IgE in the reaction system were 0nM, 0.01nM, 0.02nM, 0.05nM, 0.1nM, 0.2nM, 0.5nM, 1nM, 2nM, 5nM, 10nM, 20nM, 50nM, 100nM and 200nM, respectively.

[0079] The above-mentioned IgE37-T11-FAM is an aptamer obtained by labeling the 11th nucleotide T of the nucleic acid aptamer IgE37 with FAM.

[0080] 2. Incubate the reaction system in an ice box (4°C) for 20 minutes.

[0081] 3. The fluorescence anisotropy value (r) of IgE37-T11-FAM was measured at 25°C with a fluorometer.

[0082] The result is as figure 1 shown. figure 1 It was shown that the fluorescence anisotropy of IgE37-T11-FAM increased gradually with the increase of IgE concentration. At high concentration of IgE, the fluorescen...

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 fluorescent dye labeling aptamer for immune globulin E with sensitive fluorescence anisotropy (fluorescence polarization) response. The aptamer disclosed by the invention canbe specifically bound to immune globulin E (IgE) so as to produce an obvious fluorescence anisotropy (fluorescence polarization) signal change. A fluorescent dye fluorescein or tetramethyl rhodamineis modified to base sites of specific nucleotides of the aptamer of the IgE, so that the obtained fluorescent dye labeling aptamer maintains high affinity and is capable of producing sensitive signalresponse to the IgE. The aptamer labeled by the fluorescent dyes is capable of sensitively and rapidly detecting the IgE. The fluorescence anisotropy (fluorescence polarization) detection method is simple in operation and excellent in reproducibility.

Description

technical field [0001] The invention belongs to the field of analysis and detection, and in particular relates to a fluorescent dye-labeled nucleic acid aptamer of immunoglobulin E with sensitive fluorescence anisotropy (fluorescence polarization) response. Background technique [0002] Immunoglobulin E (IgE) is an important immune antibody related to various diseases such as allergy, atopic dermatitis, asthma, etc., and can be used as a marker for the diagnosis of some diseases. Sensitive detection of immunoglobulin E is of great significance for the detection of IgE level changes, disease diagnosis and treatment, and drug screening. [0003] Nucleic acid aptamers are single-stranded nucleic acid molecules screened from oligonucleotide libraries that can bind to target molecules with high specificity and high affinity. It does not need animals for screening, and can be synthesized and prepared by chemical methods after the sequence is known. The preparation cost is low, th...

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 Applications(China)
IPC IPC(8): C12N15/115G01N33/68G01N33/58
CPCC12N15/115C12N2310/16C12N2310/315C12N2310/334C12N2310/335C12N2310/3517G01N33/582G01N33/6854
Inventor 赵强白云龙
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com