Evaluating binding affinities by force stratification and force planning

A combination of force and scanning probe technology, applied in the direction of analyzing materials, measuring devices, instruments, etc.

Inactive Publication Date: 2004-01-14
BIOFORCE NANOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these existing methods are limited to complexes that can be bound to probes and dragged across different materials on the surface.

Method used

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  • Evaluating binding affinities by force stratification and force planning
  • Evaluating binding affinities by force stratification and force planning
  • Evaluating binding affinities by force stratification and force planning

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0031] Example description

[0032] In the present description, the term "object" is used to include any material capable of binding to a surface and capable of being detected and / or removed by probes (eg AFM probes). The object may be an inorganic molecule, an organic molecule, a biomolecule, a protein, a phage particle, a cell, etc., all of which are research objects of interest to people. The object binds or adsorbs to the surface, and is then scanned and removed, depending on how well the object binds to the surface. Objects can also be bonded to another material deposited on the surface. Additionally, the term "binding" is not limited to covalent bonding, but may include other types of molecular bonding, including adsorption, ionic bonding, and specific and non-specific molecular interactions.

[0033] The term "relatively" will be used to describe the amount of force exerted by the probe on the object to be investigated. The term "relatively" will describe the force o...

example 1

[0067] In this embodiment, the applied vertical force is measured by monitoring the deflection of the AFM probe up to and including the point at which the object 12 is removed. It will be appreciated that the lag time between the application of increased force to the object 12 and the measurement by the probe can create an amount of force that is exerted in an object that cannot be directly measured. However, the gap can be calculated by an AFM gauge or a computer. Example 1: Molecular force panning

[0068] In this example, the present invention is used to selectively remove antibodies from antigens bound to surface 10. Then, the binding force between the antibody and the antigen is measured. This example shows how the invention can be used to measure the binding force between deposited rabbit IgG and anti-rabbit IgG.

[0069] The surface in this example consisted of a glass slide covered with a layer of sputtered gold. Rabbit IgG was bound to the gold surface through non...

example 2

[0072] In other embodiments, the antigen / antibody complexes can be bound to the surface, ensuring that the entire complex is not dislodged during scans of increasing setpoints. Example 2. Force-scanning based on avidity of selection for phage and virus interactions

[0073] Such as Figure 3a As shown in -c, force saccades can alternatively be used to selectively dislodge virus or phage particles that do not bind well to surfaces, thereby allowing the collection and identification of those particles with high surface binding. Phage display is a method well known in the art for displaying the sum of potentially desired recombinant proteins on the surface of phage particles. Phages are engineered to produce specific proteins on their surfaces. Very large combinatorial libraries of proteins can be displayed on the surface of phage and selected by several screening mechanisms. Once selected, the desired phage can be propagated, and molecules with desired properties can be purif...

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Abstract

The present invention is a method for selectively removing objects from a surface utilizing a probe. The probe is scanned over the surface utilizing a greater and greater relative amount of force so that a certain number of the objects are removed from the surface. The force required to remove the objects from the surface can be calculated utilizing Hook's law and the spring constant of the probe. After removal of the objects that have a relatively weaker binding affinity with the surface, the remaining objects can be harvested, characterized, and subjected to further study.

Description

[0001] priority [0002] This application claims priority to US Provisional Patent Application 60 / 238,556, filed October 10, 2000, and US Nonprovisional Patent Application No., filed October 9, 2001. The entire disclosures of the aforementioned Provisional Application 60 / 238,556 and the aforementioned US Nonprovisional Patent Application are incorporated herein by reference. technical field [0003] The present invention is a method for analyzing objects bound to surfaces or interactions between materials on surfaces as a function of molecular forces. More specifically, the present invention is a method for distinguishing and evaluating objects by their binding affinity. Background technique [0004] Measuring the binding forces between materials, molecules, and cells is critical to a wide range of industries including, but not limited to, materials development, semiconductor production, bioanalytical assays, biomedical diagnostics, and drug development. With the advent of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 埃里克·亨德森柯蒂斯·莫舍
Owner BIOFORCE NANOSCI
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