Biomolecular sensors and methods

A sensor and molecular technology, applied in biochemical equipment and methods, nanotechnology for sensing, and microbial assay/inspection, etc., which can solve the functional barriers, inability to engineer design, sensitivity and Issues such as sensor density limitations

Active Publication Date: 2018-05-11
ROSWELL BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, carbon nanostructures present various obstacles in the functionalization of biosensors
In particular, there is no way to engineer attachment sites at specific ideal atomic positions to attach functional or sensitive probe molecules
In addition, current limitations in precision, control, and scale of carbon nanostructure synthesis pose further challenges for th...

Method used

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  • Biomolecular sensors and methods
  • Biomolecular sensors and methods
  • Biomolecular sensors and methods

Examples

Experimental program
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Effect test

Embodiment 1

[0103] Self-assembly of biopolymer bridges

[0104] The end-to-end length was constructed using an oligonucleotide of SEQ ID NO: 1 containing a 5'-thiol modification and an oligonucleotide of SEQ ID NO: 2 containing a 5'-thiol modification and an internal biotin modification A double-stranded DNA biopolymer bridge molecule of approximately 20 nm. Bridge molecules were labeled for visualization purposes using a streptavidin-gold tag. A test array 1200 of gold nanoparticle contacts was fabricated using electron beam lithography ( Figure 12 ) to deposit pairs of gold contacts, each pair of contacts defining a center-to-center contact gap of about 20 nm. A buffer solution containing gold-labeled bridging molecules was contacted with the test array of gold nanoparticle contacts. After a brief incubation period, excess solution was removed, the arrays were washed and imaged by scanning electron microscopy (SEM). Figure 12 A SEM image showing the arrangement of gold contacts 12...

Embodiment 2

[0106] Detection of self-assembly steps

[0107] A sensor device with a single sensor comprising gold contacts attached to electrodes with a center-to-center contact gap of about 20 nm was fabricated using electron beam lithography. The sensor is packaged with a PDMS flow cell that includes a 1 mm wide by 0.4 mm high channel that is open at either end to allow the introduction of liquid into the first end inside the flow cell and the discharge of liquid from the second end of the flow cell. , and the pool contacts the sensor solution. The direction of the flow cell channel is normal to the direction of the electrodes containing the sensor, which is located approximately halfway along the length of the flow cell channel. A low ionic strength buffer solution was introduced into the flow cell and a 0.5 V potential was applied to the sensor throughout the following successive steps: Introduction and self-assembly of double-stranded DNA bridge molecules (as described above in Exam...

Embodiment 3

[0114] Detection of nucleotide base incorporation

[0115] Sensor devices comprising biopolymer bridge molecules and Klenow fragment probes were fabricated and assembled as described in Example 2 above. A sensor device is used to generate signal traces in response to DNA synthesis reactions performed using single-stranded DNA templates of varying length and sequence composition. Figure 17 Signal traces for providing template sequences incorporating single bases are shown. The signal feature at 0.5 s was interpreted as corresponding to template-dependent activity and base incorporation of the Klenow fragment, and the much weaker signal feature after 0.6 s was interpreted as corresponding to some form of noise or spuriousness in the system. Signal. Figure 18 Signal traces for various template regions are shown. The template and primers described in Example 2 above were used in the reactions shown.

[0116] The top and bottom signal traces are control experiments where dNTP...

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Abstract

Electronic sensors configured to detect single molecule targets and methods of using and manufacturing such electronic sensors are disclosed. A sensor may include a first electrode and a second electrode separated by a sensor gap. The first and second electrodes can be coupled by a sensor complex that can include a biopolymer bridge molecule and a probe. The probe can interact with a target molecule, and interaction of the probe and target molecule can produce a signal suitable to provide detection of the target molecule.

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Provisional Patent Application No. 62 / 184,776, the disclosure of which is incorporated herein by reference. technical field [0003] The present disclosure relates to electronic sensor devices. In particular, the present disclosure relates to electronic sensor devices that include one or more biomolecular components in a measurement circuit. Background technique [0004] Measuring properties at the molecular scale presents many challenges due to the sensitivity required and the many potential sources of noise. Therefore, when describing a sensor for this purpose, it is helpful to be clear about all sources of measurement error. In general, for any system or object that may be measured, the measured state m is only an approximation of the actual system state a. This may be due to any of a number of factors, such as imperfect signal interpretation reflecting errors caused by the operati...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/327
CPCG01N27/4145C12Q1/6869C12Q2521/101C12Q2537/157C12Q2563/116C12Q2565/133C12Q2565/607G01N27/3272G01N27/3276B82Y15/00G01N27/3278B82Y40/00G01N27/26G01N27/327
Inventor B·L·梅里曼P·W·莫拉
Owner ROSWELL BIOTECH INC
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