Long rigid spacers to enhance binding kinetics in immunoassays

A technology of spacers and binding molecules, applied in the field of devices for target molecules

Active Publication Date: 2014-07-23
SIEMENS MEDICAL SYST NETHERLANDS LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there is not much orientation of the particle-target complex, which results in efficient binding

Method used

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  • Long rigid spacers to enhance binding kinetics in immunoassays
  • Long rigid spacers to enhance binding kinetics in immunoassays
  • Long rigid spacers to enhance binding kinetics in immunoassays

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0265] Example 1 – Using long dsDNA as an adapter

[0266] The length (extension) of the linker was analyzed in one set of experiments. For this reason, it is a very rigid molecule and thus also leads to long dsDNA extending far from the surface to serve as linkers. Specifically, 500 nm streptavidin-coated particles were incubated with dsDNA of different lengths. Each DNA molecule contains a biotin moiety at one end of the DNA molecule and a Texas Red molecule at the other end of the molecule. A solution containing particles and DNA was injected into the cartridge and bound to the sensor surface, coated with anti-Texas Red antibody using magnetic attraction. After a magnetic washing step that removes unbound particles from the surface, the number of particles bound to the surface is determined.

[0267] The binding kinetics of the particles to the surface was found to be significantly enhanced. For example, for magnetic nanoparticles with an average diameter of ~500 nm, ...

Embodiment 2

[0269] Example 2 – Effect of linker surface density on binding kinetics

[0270] The effect of linker surface density on binding kinetics was tested in an assay format using streptavidin-coated 500 nm particles that bind to a single "target molecule" that is present in A dsDNA strand with biotin at one end and a Texas Red molecule attached to the other end (see Figure 9 , left panel, diamonds), which can be used to bind to the sensor surface coated with anti-Texas Red antibody. Subsequently, different amounts of non-functional dsDNA of the same length (but lacking the Texas Red molecule) were also bound to the particles, and the ability of the particles to bind to the sensor surface was compared.

[0271] In a further experiment, particles were loaded with 1999bp dsDNA. Figure 9 , middle panel, shows the relative amount of particle binding after loading with non-functional DNA, compared to particles with only one "functional" DNA molecule.

[0272] In further experiment...

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Abstract

The present invention relates to a device for detecting a target molecule within a sample comprising a sample container for the measurement of the target molecule within a sample, a first particle, wherein said first particle is functionalized with a first binding molecule capable of specifically binding to said target molecule, and a surface structure comprising a second binding molecule, wherein said surface structure covers a flat sensor or is present on a second particle, wherein said first particle is capable of binding said second binding molecule of the surface structure directly or indirectly; wherein said first and / or second binding molecule is indirectly attached to the particle surface of said first and / or second particle and / or the flat sensor surface via a long and rigid linker molecule; wherein the length and the consistency of said linker molecule is selected such as to result in an average extension length of said linker of more than 60 nm; and wherein the number of particle clusters or of bound particles is directly or inversely related to the amount of target molecules present in the sample. In a further aspect the present invention relates to a method of detecting the presence or amount of a target molecule within a sample. The present invention also describes the use of a particle according to the invention for detecting a target molecule within a sample.

Description

technical field [0001] The present invention relates to a device for detecting target molecules in a sample comprising a sample container for measuring target molecules in a sample, a first particle, wherein said first particle is capable of specifically binding to said target molecule functionalized with a first binding molecule, and a surface structure comprising a second binding molecule, wherein the surface structure covers a flat sensor or is present on a second particle, wherein the first particle is capable of directly or indirectly binding to the surface The second binding molecule of the structure; wherein the first and / or second binding molecule is indirectly connected to the particle surface and the particle surface of the first and / or second particle via a long and rigid linker molecule and / or a flat sensor surface; wherein the length and consistency of the linker molecules are selected so as to result in an average extension length of the linker exceeding 60 nm; a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/00G01N33/52G01N33/553G01N33/543G01N33/58
CPCG01N33/54326G01N33/54353G01N33/54366G01N33/6887G01N2333/58G01N33/54393G01N33/74G01N21/552G01N21/00G01N33/553
Inventor T·H·埃弗斯M·科特斯
Owner SIEMENS MEDICAL SYST NETHERLANDS LTD
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