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Sensing device and method for detecting binding energy and binding dynamics of molecules

A technology for detecting molecules, sensing devices

Active Publication Date: 2017-02-22
DEZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has many limitations: first, the graphene functionalization process introduces more defects, which destroys the intrinsic structure of graphene, and the sensitivity and signal-to-noise ratio of the sensor are poor; second, the graphene field effect tube uses In polycrystalline graphene, the chaotic distribution of crystal domains and defects makes graphene performance uniformity poor, and the data measured by different devices are quite different, resulting in poor accuracy; Difficult, the sensor chip cannot be reused
Due to the constraints of sensitivity, signal-to-noise ratio, accuracy, and non-reproducibility, the existing sensors can only be used in a limited analysis system, and it is difficult to become a broad-spectrum, standardized analysis method.

Method used

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  • Sensing device and method for detecting binding energy and binding dynamics of molecules
  • Sensing device and method for detecting binding energy and binding dynamics of molecules
  • Sensing device and method for detecting binding energy and binding dynamics of molecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Embodiment 1: A kind of sensing device that detects binding energy and binding kinetics between molecules, such as figure 1 , 5 As shown, including sensor 2 and microfluidic chip 1;

[0086] Such as figure 2 As shown, the sensor 2 is provided with several field effect transistors, and several field effect transistors are arranged into a field effect transistor array 9, and each field effect transistor is provided with a conductive channel composed of a single-layer single-crystal graphene 12, All FET arrays form multiple parallel detection channels. Single-layer single-crystal graphene such as Figure 6 shown;

[0087] The microfluidic chip 1 is provided with a groove 7, a sample inlet 6, a sample outlet 8, and a grid inlet 5, and the sample inlet 6, the sample outlet 8, and the grid inlet 5 are arranged on the microfluidic chip 1. On the side, the sample inlet 6 and the sample outlet 8 are communicated with both ends of the groove 7 respectively;

[0088] The se...

Embodiment 2

[0093] Example 2: Affinity of DNA hybridization using the device of Example 1. The process of immobilizing the probe molecules on the graphene surface and combining the analyte molecules with the probe molecules is as follows: Figure 7 shown.

[0094] (1) A dimethylformamide (DMF) solution of 10 mM 1-pyrenebutyric acid succinamide (PBASE) was injected into the surface of the graphene single crystal through a microfluidic chip with a syringe pump, incubated at room temperature for 1 h, and injected with pure Wash off excess PBASE with DMF; inject 100mM 5' end aminated single-stranded DNA (sequence: H 2 N-(CH 2 ) 6 -5'-GAGTTGCTACAGACCTTCGT-3', code: P20) aqueous solution onto the graphene surface, incubated at room temperature for 6h, immobilized DNA probe P20 to the graphene single crystal surface;

[0095] (2) Add the DNA to be tested (sequence: 3′-CTCAACGATGTCTGGAAGCA-5′, code: T 20) into 0.01×PBS buffer to form a sample solution group to be tested (concentration: 0.25, ...

Embodiment 3

[0102] Example 3: Using the device in Example 1 to compare the difference between perfectly matched DNA hybridization and single site mismatch hybridization.

[0103] As described in Example 2, the difference is:

[0104] In step (1), the concentration of PBASE is 5 mM, and the 5' end aminated probe DNA (sequence: H2N-(CH2)6-5'-ACCAGGCGGCCGCACACGTCCTCCAT-3'; number: P26);

[0105] In step (2), the DNA to be tested is a complete match DNA (sequence: 3'-TGGTCCGCCGGCGTGTGCAGGAGGTA-5', numbering: T26) and a single site mismatch DNA (sequence: 3'-TGGTCCGCCGGCGCGTGCAGGAGGTA-5', numbering: T26( TC13)); The concentrations of the two DNA sample solutions to be tested are all 5nM;

[0106] Step (3) is with embodiment 2, and fitting result is as shown in the following table:

[0107] Table 2. P26-T26, P26-T26 (T C 13) Kinetic parameters and equilibrium constants of hybridization

[0108] k a (×10 5 m -1 the s -1 )

[0109] The results measured in this example are ...

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Abstract

The invention discloses a sensing device and method for detecting binding energy and binding dynamics of molecules. The sensing device consists of a sensor, a micro-fluidic chip and a measuring circuit. The sensor consists of a plurality of field-effect tubes, each field-effect tube adopts a single-layer graphene monocrystal as a conductive channel and has very high sensitivity and stability. The field-effect tubes are arranged in an array mode and are provided with multi-channel measuring circuits, the binding dynamics processes of different transcripts of different molecules or the same molecule can be parallelly detected, and the high-flux detection requirement is met. A compound in non-covalent binding with the single-layer monocrystal graphene is adopted as a medium to fix a probe molecule to the surface of the conductive channel, accordingly the intrinsic structure of the graphene is retained, and the signal-to-noise ratio and sensitivity of graphene field-effect tubes are improved.

Description

technical field [0001] The invention belongs to the technical field of sensing equipment and detection methods, and relates to a sensing device and method for detecting binding energy and binding kinetics between molecules. Background technique [0002] The detection of binding energy and binding kinetics between molecules has important application value in basic scientific research, screening and development of new drugs, disease diagnosis, process control of food and pharmaceutical industries, etc. Depending on whether the molecule to be tested is required to have a specific label, detection methods can be divided into two categories: label-dependent and label-independent. The latter does not require the molecule to be tested to have a specific label, but only relies on its physical properties such as molecular weight and charge for detection, which has the advantages of convenient operation and wide adaptability. At present, the detection of binding energy and binding ki...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/414
CPCG01N27/4146G01N33/523G01N33/557
Inventor 许士才詹剑王吉华周耀旗
Owner DEZHOU UNIV