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

AFM probe for single molecular force spectrum analysis and substrate functional modification method

A technology of single-molecule force spectroscopy and modification methods, which is applied in scanning probe technology, scanning probe microscopy, measuring devices, etc., and can solve problems such as complicated steps and increased risk of gold surface functionalization failure

Inactive Publication Date: 2014-02-12
SHANGHAI NAT ENG RES CENT FORNANOTECH
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage is that the steps are more complicated, thus increasing the risk of failure to functionalize the gold surface

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
  • AFM probe for single molecular force spectrum analysis and substrate functional modification method
  • AFM probe for single molecular force spectrum analysis and substrate functional modification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Functional modification of gold probes: wash the gold-plated atomic force microscope probes (gold probes) in chloroform, dry under nitrogen, wash with ultraviolet and ozone for 10 min, then wash with ultrapure water, wash with chloroform, and dry under nitrogen; The clean gold probe and gold substrate were put into 1 mM Boc-PEG-3000-SH and mPEG-2000-SH mixture (molar ratio 0.5:99.5) in chloroform for 12h, and then added to trifluoroacetic acid React in medium for 5 minutes, wash with ultrapure water, dissolve the cross-linking agent Sulfo-SMCC in ultrapure water, the concentration is 5mg / mL, dilute to 0.5mg / mL with PBS, take 100μL to react with the gold probe for 30min; dissolve SATA to two In methylformamide, the concentration is 1mg / mL, take 1.5μL SATA solution and add dropwise to 100μL 1mg / mL monoclonal anti-goat IgG, react for 2h, then take 3μL deacetylation buffer (0.5M hydroxylamine, 25mM EDTA, PBS buffer system, pH 7.2-7.5) was added dropwise to 30 μL monoclonal ...

Embodiment 2

[0029] Functional modification of gold probes: wash the gold-plated atomic force microscope probes (gold probes) in chloroform, dry under nitrogen, wash with ultraviolet and ozone for 10 min, then wash with ultrapure water, wash with chloroform, and dry under nitrogen; The clean gold probe and gold substrate were put into 1 mM Boc-PEG-3000-SH and mPEG-2000-SH mixture (molar ratio 1:99) in chloroform for 12h, and then added to trifluoroacetic acid React in medium for 5 minutes, wash with ultrapure water, dissolve the cross-linking agent Sulfo-SMCC in ultrapure water, the concentration is 5mg / mL, dilute to 0.5mg / mL with PBS, take 100μL to react with the gold probe for 30min; dissolve SATA to two In methylformamide, the concentration is 1mg / mL, take 1.5μL SATA solution and add dropwise to 100μL 1mg / mL monoclonal anti-goat IgG, react for 2h, then take 3μL deacetylation buffer (0.5M hydroxylamine, 25mM EDTA, PBS buffer system, pH 7.2-7.5) was added dropwise to 30 μL monoclonal anti...

Embodiment 3

[0033] Functional modification of gold probes: wash the gold-plated atomic force microscope probes (gold probes) in chloroform, dry under nitrogen, wash with ultraviolet and ozone for 10 min, then wash with ultrapure water, wash with chloroform, and dry under nitrogen; Clean gold probes and gold substrates were put into 1 mM Boc-PEG-3000-SH and mPEG-2000-SH mixture (molar ratio 5:95) in chloroform for 12 hours, and then added to trifluoroacetic acid React in medium for 5 minutes, wash with ultrapure water, dissolve the cross-linking agent Sulfo-SMCC in ultrapure water, the concentration is 5mg / mL, dilute to 0.5mg / mL with PBS, take 100μL to react with the gold probe for 30min; dissolve SATA to two In methylformamide, the concentration is 1mg / mL, take 1.5μL SATA solution and add dropwise to 100μL 1mg / mL monoclonal anti-goat IgG, react for 2h, then take 3μL deacetylation buffer (0.5M hydroxylamine, 25mM EDTA, PBS buffer system, pH 7.2-7.5) was added dropwise to 30 μL monoclonal a...

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 provides an AFM single molecular force spectrum measuring probe for single molecular force spectrum analysis and a substrate surface functionalization method. At first, a PEG macromolecule is coupled to an AFM probe decorated by a gold membrane or a gold substrate, one end of the PEG macromolecule is protected by a Boc group, the other end of the PEG macromolecule is a sulfhydryl group, then primary amine contained in a biomolecule is decorated to the sulfhydryl group by means of a protein modification reagent SATA or SATP, finally the Boc protection group is removed and reacts with the sulfhydrylation biomolecule, and therefore protein is decorated to the gold probe and the gold substrate. The method is simple in process and easy and convenient to operate, and targets of reaching interaction force between biomolecules and cell surface single molecular recognition and the like in the single molecule level can be achieved.

Description

technical field [0001] The invention relates to a probe and a substrate functional modification method, in particular to an atomic force microscope probe for single-molecule force spectrum analysis and a substrate functional modification method, belonging to the field of microscience, in particular to nanometer measurement. Background technique [0002] Today, atomic force microscopy single-molecule force spectroscopy has become a powerful tool for studying the interactions between biomolecules. A cutting-edge direction in the interdisciplinary fields of chemistry, chemistry and physics. In the study of single-molecule force spectroscopy, the protein is usually fixed on the tip of the atomic force microscope probe and the substrate through a polyethylene glycol (PEG) polymer chain, so that the spatial freedom of biomolecules can be guaranteed, and the force-distance The graph separates specific interactions between biomolecules from non-specific interactions between the tip...

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
IPC IPC(8): G01Q60/38G01Q60/24
Inventor 钟建马梦佳魏岱旭闫志强周涓李文英何丹农
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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