CN-115792102-B - Multi-parameter integrated bionic odor sensing chip and preparation method and application thereof

CN115792102BCN 115792102 BCN115792102 BCN 115792102BCN-115792102-B

Abstract

The invention discloses a multiparameter integrated bionic odor sensing chip, a preparation method and application thereof, and the sensing chip is characterized in that tens of human olfactory receptor proteins and field effect transistors are integrated on the same chip by combining MEMS technology and bionic sensing technology, and the sensing chip has the characteristics of high flux, small size, high integration level and low energy consumption. After the gas molecules are combined with the olfactory receptor proteins on the chip, protein structure phase change is caused, the grid current of the field effect transistor is changed, the current signals are normalized, the response patterns of different gas molecules are constructed to form an odor database, and the high-dimensional detection and accurate identification of a large number of different odors can be realized by utilizing the cross sensitivity of the olfactory receptor proteins and combining a pattern recognition algorithm. The odor sensing chip provided by the invention has wide application prospects in the fields of air quality monitoring, food safety, hazard detection and the like.

Inventors

PAN YUXIANG
WANG DI
XU FENG
CHENG CHEN

Assignees

之江实验室

Dates

Publication Date: 20260508
Application Date: 20221031

Claims (4)

1. A multiparameter integrated bionic odor sensing chip is characterized by comprising a plurality of mutually independent graphene FET sensors, wherein each graphene FET sensor comprises a silicon oxide substrate, a source electrode, a drain electrode, a graphene gate electrode, a source lead, a drain lead, a gate lead, an olfactory receptor protein and a bonding pad, the source electrode, the drain electrode and the graphene gate electrode are attached to the silicon oxide substrate, the graphene gate electrode is positioned between the source electrode and the drain electrode, the three electrodes are respectively connected with the bonding pad through the corresponding source lead, the drain lead and the gate lead, olfactory receptor proteins are fixed on the surface of the graphene gate electrode, the olfactory receptor proteins are human olfactory receptor proteins, the olfactory receptor proteins are respectively modified on the mutually independent graphene gate electrodes of the FET sensors, the olfactory receptor proteins are OR1A1、OR1A2、OR1A3、OR1D2、OR1G1、OR2B11、OR2C1、OR2J2、OR2J3、OR2M4、OR2W1、OR3A1、OR5B17、OR10A6、OR10G7、OR52D1; as graphene FET sensitive materials of the sensors, and are subjected to conformational change after being combined with gas molecules, so that the FET sensors are changed, the response characteristics of different proteins are different from each other, the FET devices have cross specific recognition capability, and have different current response to different current chips, have different gas-phase response patterns, and different gas-response patterns are different to different gas-phase response patterns, and different gas-response patterns are realized, and different gas-like recognition patterns are realized: The odor response heat map construction comprises the steps of testing hundreds of known odors, extracting signal characteristics [ I, n ] of each FET sensitive unit after different types of odors are processed, wherein I is a peak current signal value, n is the number of the FET sensitive unit, spatial position information of a graphene FET sensor is contained, and the peak current signal is converted into a visual response heat map to form an odor response map database; And (3) odor identification, namely extracting signal characteristics [ I, n ] of each FET sensitive unit after unknown gas reacts, converting the signal characteristics into a one-dimensional array, and comparing the Euclidean distance with the Euclidean distance by calculating the Euclidean distance of the array and an odor response database and selecting the closest value to realize odor identification.
2. The multi-parameter integrated bionic odor sensing chip of claim 1 wherein the source and drain electrodes are gold and the gate electrode is graphene.
3. A method for preparing a multi-parameter integrated bionic odor sensing chip according to any one of claims 1-2, comprising the steps of: s1, adopting a silicon wafer as a substrate, and carrying out organic cleaning on the silicon wafer; s2, preparing a silicon oxide layer on the surface of the silicon wafer; S3, transferring the graphene film onto a silicon oxide substrate; S4, etching a grid pattern on the graphene film to serve as a graphene grid of the FET device; s5, manufacturing a source electrode, a drain electrode, a source electrode wire, a drain electrode wire and a bonding pad on the silicon oxide layer; s6, forming a silicon oxide insulating layer on the source electrode wire and the drain electrode wire; s7, dicing and packaging the silicon wafer; S8, expressing human olfactory receptor protein by adopting a cell-free system method; S9, purifying olfactory receptor proteins by adopting a protein purification device; S10, constructing phospholipid vesicles by using a micro extruder, mixing and transferring olfactory receptor proteins, the phospholipid vesicles and Brij35 into a dialysis membrane for dialysis for 3-4 days, and centrifuging for 60 minutes to collect three-dimensional reconstructed olfactory receptor proteins; s11, dripping the recombined olfactory receptor protein on the polylysine treated FET array graphene gate electrode 5, and completing interface coupling of the olfactory receptor protein and a graphene FET chip through electrostatic adsorption.
4. Use of a multiparameter integrated biomimetic odour-sensing chip according to any one of claims 1-2 for odour detection, comprising the steps of: (1) The signal detection, namely cleaning the graphene FET sensors by using ultrapure water, drying the graphene FET sensors by using N 2 , dripping 20 mu L of odor buffer solution to be detected on the surface of each independent graphene FET sensor, connecting a bonding pad with a source meter by using a lead, and recording a voltage-current characteristic curve through a Keithley 2636A source meter; (2) The odor response heat map construction comprises the steps of testing hundreds of known odors, extracting signal characteristics [ I, n ] of each FET sensitive unit after different types of odors are processed, wherein I is a peak current signal value, n is the number of the FET sensitive unit, spatial position information of a graphene FET sensor is contained, and the peak current signal is converted into a visual response heat map to form an odor response map database; (3) And (3) odor identification, namely extracting signal characteristics [ I, n ] of each FET sensitive unit after unknown gas reacts, converting the signal characteristics into a one-dimensional array, and comparing the Euclidean distance with the Euclidean distance by calculating the Euclidean distance of the array and an odor response database and selecting the closest value to realize odor identification.

Description

Multi-parameter integrated bionic odor sensing chip and preparation method and application thereof Technical Field The invention relates to the technical field of bionic odor detection, in particular to an odor sensing chip based on human olfactory receptor protein and FET sensor arrays. Background The gas detection technology has great economic and social benefits in the fields of food safety, industrial production, national defense safety and medical health, and the core component of the gas detection technology is a gas sensor chip. Conventional gas sensors include semiconductor, electrochemical, catalytic combustion, thermal conductivity, and infrared gas sensors. The existing gas sensor has the problems of few sensitive elements, high energy consumption, poor selectivity, low integration level and the like, and is limited to be widely applied to the fields of industrial Internet of things, security protection, military industry, food and the like. Human olfactory receptor proteins are chemoreceptors expressed on the membrane of the olfactory receptor neurons and are responsible for detecting odors that produce olfaction. The human olfactory system is composed of more than 400 olfactory receptor proteins, and recognition of millions of odors can be achieved by virtue of the cross-specificity of the olfactory receptor proteins. The human olfactory receptor protein is used as the sensitive element of the bionic odor sensing chip, so that the human olfactory system can be restored to the greatest extent in vitro. Tens of olfactory receptor proteins are covalently coupled with a field effect transistor array to construct a bionic odor sensing chip, so that the detection range, accuracy and sensitivity of a traditional gas sensor can be effectively improved. The high-dimensional detection and accurate identification of complex smell are realized, and the sense of smell reality of human beings is further simulated. Disclosure of Invention The invention aims to solve the technical problem of providing a gas sensor array based on human olfactory receptor protein and a manufacturing method thereof, wherein the gas sensor array has the advantages of multiple detection parameters, high integration level, high sensitivity and mass production. The invention provides a multi-parameter integrated bionic odor sensing chip, which comprises a plurality of mutually independent graphene FET sensors, wherein each graphene FET sensor comprises a silicon oxide substrate, a source electrode, a drain electrode, a graphene gate electrode, a source electrode lead, a drain electrode lead, a grid lead, an olfactory receptor protein and a bonding pad, the source electrode, the drain electrode and the graphene gate electrode are attached to the silicon oxide substrate, the graphene gate electrode is positioned between the source electrode and the drain electrode, the three electrodes are respectively connected with the bonding pad through the corresponding source electrode, the drain electrode and the grid lead, the olfactory receptor protein is fixed on the surface of the graphene gate electrode, the olfactory receptor protein is used as a graphene FET sensitive material of the sensor, and is subjected to conformational change after being combined with gas molecules, so that the leakage current of the FET sensor is changed, each FET device has different current responses, the chip has different response heat maps for different gases, and the detection algorithms of different types of gases are realized through combination mode identification. Further, the source electrode and the drain electrode are made of gold, and the gate electrode is made of graphene; further, the olfactory receptor protein is human olfactory receptor protein, and is respectively modified on the graphene grid electrodes of the FET sensors which are mutually independent. In a second aspect, the invention also provides a preparation method of the bionic odor sensing chip based on multi-parameter integration, which comprises the following steps: s1, adopting a silicon wafer as a substrate, and carrying out organic cleaning on the silicon wafer; s2, preparing a silicon oxide layer on the surface of the silicon wafer; S3, transferring the graphene film onto a silicon oxide substrate; S4, etching a grid pattern on the graphene film to serve as a graphene grid of the FET device; s5, manufacturing a source electrode, a drain electrode, a source electrode wire, a drain electrode wire and a bonding pad on the silicon oxide layer; s6, forming a silicon oxide insulating layer on the source electrode wire and the drain electrode wire; s7, dicing and packaging the silicon wafer; S8, expressing human olfactory receptor protein by adopting a cell-free system method; S9, purifying olfactory receptor proteins by adopting a protein purification device; S10, constructing phospholipid vesicles by using a micro extruder, mixing and transferring olfactory receptor proteins, the phosp