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Method and apparatus for detecting molecular binding events

a molecular binding and event technology, applied in biochemistry apparatus and processes, instruments, material analysis, etc., can solve the problems of detection methods, detection methods, and difficult or even impossible to label one or all of the molecules needed for a particular assay,

Inactive Publication Date: 2002-06-13
MDS SCIEX
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, in many cases it is difficult or even impossible to label one or all of the molecules needed for a particular assay.
In addition, none of these labeling approaches determines the exact nature of the binding event, so for example active site binding to a receptor is indistinguishable from non-active-site binding such as allosteric binding, and thus no functional information is obtained via the present detection methodologies.
This configuration has several drawbacks, including some substantial limitations on the frequencies useable in the detection strategy, and a profound limitation on the sensitivity of detecting molecular systems.
In general, limitations exist in the areas of specificity and sensitivity of most assay systems.
Cellular debris and non-specific binding often cause the assay to be noisy, and make it difficult or impossible to extract useful information.
As mentioned above, some systems are too complicated to allow the attachment of labels to all analytes of interest, or to allow an accurate optical measurement to be performed.
Further, a mentioned above, most of these detection technologies yield no information on the functional nature of the binding event.

Method used

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  • Method and apparatus for detecting molecular binding events
  • Method and apparatus for detecting molecular binding events
  • Method and apparatus for detecting molecular binding events

Examples

Experimental program
Comparison scheme
Effect test

second embodiment

[0127] FIG. 1B illustrates the bio-assay system comprising an array of resonant microstrip circuits 170. Each resonant circuit 170 consists of a transmission line 172 terminating in an open-circuited stub 176. Those skilled in the art of circuit design will appreciate other resonant structures may be employed in lumped element, distributed, or a combination of both circuit topologies.

[0128] FIG. 1C illustrates a cross-section view of one resonant circuit 170. The open-circuited stub 176 forms the bio-electrical interface of the resonant circuit 170 and closely parallels the bio-electrical interface shown in FIG. 1A. In particular, the open-circuited stub 176 consists of an interface transmission line 176a deposited on a dielectric layer 176b, and is positioned above ground plane 176c.

[0129] In this embodiment, the MBL 176d is coupled via a direct connection to transmission line 176a. The MBL 176d can bind along the interface transmission line in a specific or non-specific manner. As...

example 1

A. Example 1

Detection of a Ligand Binding to the Surface.

[0282] Primary binding of urease to an ITO surface was demonstrated in the bio-assay device as shown in FIG. 2A. The binding surface of the bio-assay device comprised a cover glass treated with ITO deposited via chemical vapor deposition (CVD). The ITO transmission line was carefully examined to ensure that it contained no microfractures or breaks in it. The transmission line was measured with a Tektronix 11801 signal analyzer with a TDR module, and found to have a broadband reference impedance of 32 .OMEGA.. The line length was about 2.6 nsec in length, the binding surface was found to have an impedance of 34 .OMEGA., and a length of about 200 psec. Separation between the top and bottom plates were 10 mils, and the chamber was 1 / 2" long. No side walls were used; instead, the capillary action of the top and bottom plates retained the solution in place.

[0283] Next, the bio-assay device filed with a solution of d-PBS. With the b...

example 2

B. Example 2

Identification of Collagenase and Lysozyme through Primary Binding

[0287] Using a bio-assay device similar to the one cited in example 1 above, and prepared and characterized in a similar manner, we carried out a series of experiments to examine the differing responses of different proteins over the frequency range of 1-10GHz. The same device was used for each experiment (to eliminate small differences in fabrication from one device to another), but was thoroughly washed with SDS between the application of each of the proteins. FIGS. 12C and 12D illustrate the transmission loss measurements of the primary binding effects of collagenase and lysozyme samples, respectively, over the test frequency range from 1 GHz to 10 GHz. In both instances, the signal response exhibited a pattern of peaks and valleys which can be used to detect and identify the ligand uniquely. In particular, the frequency response of the collagnase sample exhibited a strong positive peak near 5 GHz. The ...

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Abstract

Systems and methods for detecting molecular binding events and other environmental effects using the unique dielectric properties of the bound molecular structure or structures are presented. A molecular binding layer is coupled along the surface of a signal path. A test signal is propagated along the signal path, whereby the test signal couples to the molecular binding layer, and in response, exhibits a signal response.

Description

BACKGROUND OF THE INVENTION[0001] Virtually every area of biomedical sciences is in need of a system to assay chemical and biochemical reactions and determine the presence and quantity of particular analytes. This need ranges from the basic science research lab, where biochemical pathways are being mapped out and their functions correlated to disease processes, to clinical diagnostics, where patients are routinely monitored for levels of clinically relevant analytes. Other areas include pharmaceutical research, military applications, veterinary, food, and environmental applications. In all of these cases, the presence and quantity of a specific analyte or group of analytes, needs to be determined.[0002] For analysis in the fields of chemistry, biochemistry, biotechnology, molecular biology and numerous others, it is often useful to detect the presence of one or more molecular structures and measure binding between structures. The molecular structures of interest typically include, b...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/543
CPCG01N33/54373
Inventor HEFTI, JOHN
Owner MDS SCIEX