CN110596401A - High-field asymmetric waveform ion mobility device and method for protein detection - Google Patents
High-field asymmetric waveform ion mobility device and method for protein detection Download PDFInfo
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- 238000002331 protein detection Methods 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 title claims description 19
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 77
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 77
- 238000000926 separation method Methods 0.000 claims abstract description 47
- 238000000137 annealing Methods 0.000 claims abstract description 37
- 238000001514 detection method Methods 0.000 claims abstract description 17
- 150000002500 ions Chemical class 0.000 claims description 117
- 239000012159 carrier gas Substances 0.000 claims description 30
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 22
- 239000007789 gas Substances 0.000 claims description 19
- 229910052757 nitrogen Inorganic materials 0.000 claims description 11
- 239000001257 hydrogen Substances 0.000 claims description 10
- 229910052739 hydrogen Inorganic materials 0.000 claims description 10
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims description 10
- 229910018503 SF6 Inorganic materials 0.000 claims description 9
- 239000001307 helium Substances 0.000 claims description 9
- 229910052734 helium Inorganic materials 0.000 claims description 9
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims description 9
- 238000001819 mass spectrum Methods 0.000 claims description 9
- SFZCNBIFKDRMGX-UHFFFAOYSA-N sulfur hexafluoride Chemical compound FS(F)(F)(F)(F)F SFZCNBIFKDRMGX-UHFFFAOYSA-N 0.000 claims description 9
- 229960000909 sulfur hexafluoride Drugs 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 8
- 150000004945 aromatic hydrocarbons Chemical class 0.000 claims description 3
- 150000002576 ketones Chemical class 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- 150000001298 alcohols Chemical class 0.000 claims description 2
- 150000002170 ethers Chemical class 0.000 claims description 2
- 230000035945 sensitivity Effects 0.000 abstract description 7
- 238000000132 electrospray ionisation Methods 0.000 abstract description 3
- 239000000543 intermediate Substances 0.000 abstract description 3
- 230000000694 effects Effects 0.000 description 11
- 238000004140 cleaning Methods 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000005684 electric field Effects 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000006317 isomerization reaction Methods 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001871 ion mobility spectroscopy Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6848—Methods of protein analysis involving mass spectrometry
Abstract
The invention discloses a high-field asymmetric waveform ion mobility device for protein detection, which is characterized in that: the electrospray ionization source comprises an electrospray ionization source, a first group of electrode plates, a second group of electrode plates, a mass spectrometer, a sine wave power supply, a high-field asymmetric waveform power supply and a direct current power supply, wherein a sample inlet is formed in one side of the first group of electrode plates, the first group of electrode plates and the second group of electrode plates are coaxially and parallelly arranged in front and at the back, a pre-annealing area is formed between the first group of electrode plates, a separation area is formed between the second group of electrode plates, the rear end of the second group of electrode plates is connected with the mass spectrometer, the first group of electrode plates are connected with the sine wave power supply and a temperature control module, the second group of electrode plates are connected with the high-field asymmetric waveform power supply and. The invention is suitable for detecting molecules with complex structures of stereoisomers and intermediates, particularly protein, and has the advantages that: the resolution and sensitivity of detection are improved, and the repeatability is good.
Description
Technical Field
The invention relates to the technical field of ion separation detection, in particular to a high-field asymmetric waveform ion mobility device and method for protein detection.
Background
High Field Asymmetric Ion Mobility Spectrometry (FAIMS) is a rapid gas phase Ion separation technique developed from conventional Ion Mobility techniques. The mobility of the ions with the high-field asymmetric waveform utilizes the characteristic that the periods of high and low electric fields of the ions are changed alternately, and different ions are separated in the polar plate gap according to the difference of the ion mobility under the conditions of the high and low electric fields. The technology has the advantages of simple structure, small core devices, easiness in continuous detection and the like, has great development potential in the field of field analysis and detection, and is applied to separation and detection of part of protein isomers at present.
The existing high-field asymmetric waveform ion mobility device generally comprises an ionization region, a separation region and a detection region, wherein a sample is ionized into sample ions in the ionization region and then is carried into the separation region by carrier gas, the separation region is used for separating target ions by applying high-field asymmetric waveform voltage, compensation voltage is applied to compensate high-field and low-field motion deviation of the target ions, and the target ions finally enter the detection region to be detected.
The technology is still lack of the following when used for detecting molecules with complex structures such as protein and the like: (1) the resolution is low: because the structure of the protein is complex, the isomers are numerous, and the difference of the three-dimensional structure exists, the resolution of the existing high-field asymmetric waveform ion mobility technology on protein analysis is insufficient, the spatial information is difficult to determine, and the separation and identification of the stereoisomer at the complete protein level are realized; (2) the sensitivity is low: the existing high-field asymmetric waveform ion mobility technology can only pass ions of corresponding compensation voltage at the same time, and other ions are neutralized by upper and lower electrode plates of a separation area, so that ions detected by a subsequent detector are greatly reduced, and the sensitivity is influenced; (3) poor repeatability: because the structure of the protein is easily affected by temperature, the excessive temperature in the separation zone during continuous working can easily cause the protein to generate a self-cleaning phenomenon in the separation process, namely, the intermediate between more stable isomers is eliminated, and simultaneously, the signal can be seriously inhibited, thereby affecting the repeatability of sample detection.
Disclosure of Invention
In order to solve the defects in the prior art, the invention provides the high-field asymmetric waveform ion mobility device and the method for protein detection, which can improve the resolution and sensitivity of protein detection and have good repeatability.
The technical scheme adopted by the invention for solving the technical problems is as follows: a high-field asymmetric waveform ion mobility device for protein detection comprises an electrospray ion source, a first group of electrode plates, a second group of electrode plates, a mass spectrometer, a sine wave power supply, a high-field asymmetric waveform power supply, a direct current power supply and a gas generator, wherein one side of the first group of electrode plates is provided with a sample inlet for protein sample ions generated by ionization of the electrospray ion source to enter, the first group of electrode plates and the second group of electrode plates are coaxially and parallelly arranged in front of and behind, a gap is reserved between the first group of electrode plates and the second group of electrode plates, a pre-annealing area is formed between the first group of electrode plates, a separation area is formed between the second group of electrode plates, the rear end of the second group of electrode plates is connected with a detection inlet of the mass spectrometer, and the first group of electrode plates is electrically connected with the sine wave power supply, the outer side of the first group of electrode plates is provided with a temperature control module for controlling the temperature of the pre-annealing area, the second group of electrode plates is electrically connected with the high-field asymmetric waveform power supply and the direct-current power supply, the gas generator is arranged at the front end of the first group of electrode plates, and the generated carrier gas pushes ionized protein sample ions to sequentially pass through the pre-annealing area and the separation area.
In some embodiments, the first group of electrode plates and the second group of electrode plates are both formed by two plate-type electrode plates which are symmetrical up and down, and the electrode plate spacing between the first group of electrode plates and the second group of electrode plates is equal. Therefore, the method has better protein ion pre-annealing and separating effects.
In some embodiments, the distance between the electrode plates of the first group of electrode plates and the second group of electrode plates is 0.1mm to 10cm, preferably 1mm to 5 mm. Therefore, the method has a better effect and is beneficial to improving the detection resolution.
In some embodiments, the sine wave power supply has a voltage amplitude of no less than 3 kV. Therefore, the annealing reaction of the protein sample ions in the pre-annealing area can be better realized, and the performance stability of the protein ions in the subsequent separation process is improved.
In some embodiments, the high-field asymmetric waveform power supply can adopt a square wave, a half sine wave or a double sine wave, the voltage amplitude of the high-field asymmetric waveform power supply is not less than 3kV, and the frequency is 1 kHz-100 MHz, preferably 100 kHz. Thereby having a superior separation effect.
A method for detecting protein by adopting a high-field asymmetric waveform ion mobility device comprises the following steps:
firstly, using an electrospray ion source to ionize a target protein sample in a solution to generate protein sample ions, wherein the protein sample ions enter a first group of electrode plates and are pushed by carrier gas to move forwards;
providing a sine wave power supply for the first group of electrode plates, providing a stable temperature field for a pre-annealing area of the first group of electrode plates through a temperature control module, and oscillating the protein sample ions in the pre-annealing area under the combined action of the carrier gas, the sine wave power supply and the temperature field to finish pre-annealing;
thirdly, a high-field asymmetric waveform power supply and a direct-current power supply are provided for the second group of electrode plates, protein sample ions continue to move forward to a separation area in the second group of electrode plates under the pushing of carrier gas, target protein ions are separated in the separation area and enter a mass spectrometer for detection, a mass spectrum signal of scanning compensation voltage is obtained, and the rest ions are neutralized in the separation area;
changing the compensation voltage of the direct current power supply, and repeating the steps to obtain mass spectrum signals under different compensation voltages;
integrating mass spectrum signals under different compensation voltages to obtain a signal spectrogram of the high-field asymmetric waveform ion mobility of the target protein ions.
In some embodiments, the electrospray ion source produces protein sample ions including protein positive and negative ions and other ionizable particles. The electrospray ion source can ensure that the spatial structure of the generated protein sample ions is unchanged.
In some embodiments, the carrier gas is one of hydrogen, helium, nitrogen and sulfur hexafluoride, or a mixture of a plurality of hydrogen, helium, nitrogen and sulfur hexafluoride in any proportion, or a mixture of several of hydrogen, helium, nitrogen and sulfur hexafluoride and a doping gas, the doping gas is selected from ethers, alcohols, ketones and aromatic hydrocarbon gases, and the flow rate of the carrier gas is 0.001L/min to 30L/min, preferably 1L/min to 5L/min. Therefore, the carrier gas is the mixture of a plurality of hydrogen, helium, nitrogen and sulfur hexafluoride and the doping gas, and the pre-annealing and separation effects can be further improved by combining the specific flow rate.
In some embodiments, in the second step, the voltage amplitude of the sine wave power supply is not less than 3kV, and the temperature range of the temperature field is 0-500 ℃, preferably 100-300 ℃. Therefore, a stable temperature field is provided for protein ion pre-annealing, the performance of the protein ions after pre-annealing can be better achieved, the self-cleaning phenomenon during subsequent separation is reduced, and the ion signal intensity is ensured.
In some embodiments, the adjustable range of the compensation voltage of the dc power supply is-200V to 200V.
Compared with the prior art, the invention has the advantages that:
(1) the electrospray ionization source is arranged to generate protein sample ions, the protein sample ions sequentially pass through the two groups of electrode plates under the pushing of carrier gas, the protein sample ions oscillate under the action of a high-voltage sine wave power supply and a temperature field of the first group of electrode plates and collide with carrier gas molecules to convert heat energy, the temperature of the sample ions is raised to preheat, then the protein sample ions can automatically maintain the state of a potential low point, and an annealing effect is performed in a pre-annealing area, so that the space structure and the performance of protein can be stabilized, the generation of a protein self-cleaning phenomenon can be reduced, the uncontrollable self-cleaning phenomenon simultaneously occurring in the original separation process can be controlled, the subsequent separation effect is ensured, and the ion signal intensity is improved;
(2) compared with the traditional FAIMS structure, the distance between the electrode plates can be increased from 1.9mm to 5mm, the separation time can be increased from 0.2-0.4 s to 1-1.5 s, and the resolution can be increased from 200-400 to about 800 in a larger range on the premise of no sensitivity loss, so that the accurate identification of the stereoisomer at the complete protein level is realized;
(3) the duration of the protein ions in the pre-annealing stage and the separation stage can be reasonably controlled, and the protein ions after pre-annealing are not influenced by overhigh temperature in continuous working, so that further isomerization is avoided, and the repeatability of a detection map is improved.
Drawings
FIG. 1 is a schematic structural diagram of a high-field asymmetric waveform ion mobility device for protein detection according to the present invention.
The system comprises an electrospray ion source 1, a first group of electrode plates 2, a second group of electrode plates 3, a mass spectrometer 4, a sine wave power supply 5, a high-field asymmetric waveform power supply 6, a direct current power supply 7, carrier gas 8, a sample inlet 9, a pre-annealing area 10, a separation area 11 and a temperature control module 12.
Detailed Description
The high-field asymmetric waveform ion mobility device and method for protein detection according to the present invention will be described in further detail with reference to the accompanying drawings, but the invention is not limited thereto.
Example one
As shown in the figure, the high-field asymmetric waveform ion mobility device for protein detection comprises an electrospray ion source 1, a first group of electrode plates 2, a second group of electrode plates 3, a mass spectrometer 4, a sine wave power supply 5, a high-field asymmetric waveform power supply 6, a direct current power supply 7 and a gas generator, wherein one side of the first group of electrode plates 2 is provided with a sample inlet 9 for protein sample ions generated by ionization of the electrospray ion source to enter, the first group of electrode plates 2 and the second group of electrode plates 3 are coaxially arranged in parallel in the front and at the back, a gap is reserved between the first group of electrode plates 2 and the second group of electrode plates 3, a pre-annealing area 10 is formed between the first group of electrode plates 2, a separation area 11 is formed between the second group of electrode plates 3, the rear end of the second group of electrode plates 3 is connected with a detection inlet of the mass spectrometer 4, the first group of electrode plates 2 is electrically connected with the sine wave power, the second group of electrode plates 3 is electrically connected with a high-field asymmetric waveform power supply 6 and a direct current power supply 7, the gas generator is arranged at the front end of the first group of electrode plates 2, and the generated carrier gas 8 pushes ionized protein sample ions to sequentially pass through a pre-annealing area 10 and a separation area 11.
In this embodiment, the upper electrode plate of the first group of electrode plates 2 is provided with a sample inlet 9, and the sample introduction direction of the protein sample ions generated by the ionization of the electrospray ion source and the carrier gas direction form a certain angle, so that the loss of the sample ions can be reduced better.
In this embodiment, the temperature control module 12 includes a heating plate and a temperature sensor tightly disposed at the periphery of the first group of electrode plates, the heating plate is connected to an external power supply, and the temperature sensor is connected to the control system for accurately controlling the temperature of the pre-annealing region and ensuring a stable temperature field. The gas generator is also connected with a gas flow controller for controlling the flow rate of the carrier gas to be kept stable.
In this embodiment, the first group of electrode plates 2 and the second group of electrode plates 3 are both formed by two vertically symmetric flat plate-type electrode plates, the electrode plate spacing between the first group of electrode plates 2 and the second group of electrode plates 3 is equal to each other, and is 4mm, and in other embodiments, the electrode plate spacing may be 0.1mm, 1mm, 3mm, 5mm, 1cm, 10cm, and the like.
In this embodiment, the voltage amplitude of the sine wave power supply 5 is not less than 3 kV. The high-field asymmetric waveform power supply 6 is formed by superposing double sine waves, wherein the waveform formed by superposition meets the condition that the integral of one period is 0, namely the integral areas of a high-field part and a low-field part are the same, and the high-field asymmetric waveform power supply 6 can realize an approximate effect by adopting square waves, half sine waves or other high-order fitting waveforms in other embodiments. The amplitude of the voltage of the high-field asymmetric waveform power supply 6 is not less than 3kV, and the frequency is 100kHz, in other embodiments, the frequency of the high-field asymmetric waveform power supply 6 may be 1kHz, 10kHz, 500kHz, 10MHz, 100MHz, and the like.
The mass-to-charge ratio (m/z) of the mass spectrometer 4 is in the range of 1-1000000 amu, preferably 1000-200000 amu, and the mass resolution of the mass spectrometer 4 is in the range of 100-20000000. In this embodiment, the mass resolution of the mass spectrometer is not less than 10000 under the conditions of the mass-to-charge ratio of 1000amu and the highest sensitivity of the mass spectrometer, and the mass spectrometer has a better effect.
Example two
The invention relates to a method for detecting protein by adopting a high-field asymmetric waveform ion mobility device, which comprises the following steps of:
firstly, using an electrospray ion source to ionize a target protein sample in a solution to generate protein sample ions, ensuring that the spatial structure of the protein sample ions is unchanged, and enabling the protein sample ions to enter a first group of electrode plates and to move forwards under the pushing of carrier gas;
providing a sine wave power supply for the first group of electrode plates, providing a stable temperature field for a pre-annealing area of the first group of electrode plates through a temperature control module, and oscillating the protein sample ions in the pre-annealing area under the combined action of the carrier gas, the sine wave power supply and the temperature field to finish pre-annealing;
thirdly, a high-field asymmetric waveform power supply and a direct-current power supply are provided for the second group of electrode plates, protein sample ions continue to move forward to a separation area in the second group of electrode plates under the pushing of carrier gas, target protein ions are separated in the separation area and enter a mass spectrometer for detection, a mass spectrum signal of scanning compensation voltage is obtained, and the rest ions are neutralized in the separation area;
changing the compensation voltage of the direct current power supply, and repeating the steps to obtain mass spectrum signals under different compensation voltages;
integrating mass spectrum signals under different compensation voltages to obtain a signal spectrogram of the high-field asymmetric waveform ion mobility of the target protein ions.
The protein sample ions produced by the electrospray ion source include protein positive and negative ions and other ionizable particles.
The carrier gas is one or a mixture of a plurality of hydrogen, helium, nitrogen and sulfur hexafluoride according to any proportion, or the mixture of a plurality of hydrogen, helium, nitrogen and sulfur hexafluoride and doping gas, and the doping gas is selected from ether, alcohol, ketone and aromatic hydrocarbon gas, thereby improving the separation effect of the target sample. The flow rate of the carrier gas is 0.001L/min to 30L/min, preferably 1L/min to 5L/min. In the second step, the voltage amplitude of the sine wave power supply is not less than 3kV, the temperature range of the temperature field is 0-500 ℃, and the temperature is preferably 100-300 ℃.
In the embodiment, the carrier gas is pure nitrogen or a mixture of 75% nitrogen and 25% hydrogen, the flow rate is 2-3L/min, and the pre-annealing temperature is 100-150 ℃. The adjustable range of the compensation voltage of the direct current power supply is-200V. Based on the specific carrier gas flow rate, the pre-annealing temperature and the sine wave voltage, the protein is pre-annealed before separation, the uncontrollable self-cleaning phenomenon which simultaneously occurs in the original separation process is controlled, and the signal intensity of protein detection is ensured.
The invention relates to a high-field asymmetric waveform ion mobility device and a method for protein detection, which have the following basic working principles: protein sample ions are ionized by an electrospray ion source 1 to form sample ions, and then sequentially pass through two groups of flat plate electrode plates under the pushing of a carrier gas, the sample ions are subjected to the action of a stable temperature field of a high-voltage sine wave power supply 5 and a temperature control module 12 between a first group of electrode plates 2 (a pre-annealing area 10), the sample ions oscillate and collide with carrier gas molecules, partial kinetic energy is converted into heat energy, the temperature of the sample ions is increased to be preheated, and then the protein sample ions can be automatically maintained in a low-potential punctiform state to perform an annealing effect. The process can reduce the generation of protein 'self-cleaning', is favorable for stabilizing the spatial structure and performance of the protein, is favorable for subsequent separation, and improves the ionic signal intensity. Then the protein sample ions are separated by the action of the high-field asymmetric waveform power supply 6 between the second group of electrode plates 3 (separation area 11), the target ions fly out of the separation area by the compensation action of the direct current power supply 7 and are finally detected by the mass spectrometer 4, and the rest ions impact on the electrode plates to be neutralized.
Based on the characteristics, the high-field asymmetric waveform ion mobility device and the method for detecting the protein have the advantages that the protein pre-annealing stage is introduced, the sine wave with higher average voltage is adopted to pre-anneal the protein before separation, the space structure and the performance of the protein are stabilized, the generation of the self-cleaning phenomenon of the protein can be reduced, the uncontrollable self-cleaning phenomenon simultaneously generated in the original separation process can be controlled, the subsequent separation effect is ensured, and the ion signal intensity is improved; compared with the traditional FAIMS structure, the distance between the electrode plates can be increased from 1.9mm to 5mm, the separation time is increased from 0.2-0.4 s to 1-1.5 s, the residence time of ions in the separation area is prolonged, the resolution can be increased from 200-400 to about 800 on the premise of no sensitivity loss, and the accurate identification of the complete protein-level stereoisomer is realized; the duration of the protein ions in the pre-annealing stage and the separation stage can be reasonably controlled through the length of the electrode plate and the flow rate of the carrier gas, and the protein ions are not affected by overhigh temperature in continuous working after the pre-annealing, so that the further isomerization can be avoided, and the repeatability of a detection map is improved.
The high-field asymmetric waveform ion mobility device and the method for detecting the protein are also suitable for detecting other molecules with complicated structures such as stereoisomers and intermediates.
It should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and the scope of the present invention is not limited thereby, and the present invention may be modified in materials and structures, or replaced with technical equivalents, in the constructions of the above-mentioned various components. Therefore, structural equivalents made by using the description and drawings of the present invention or by directly or indirectly applying to other related arts are also encompassed within the scope of the present invention.
Claims (10)
1. A high-field asymmetric waveform ion mobility device for protein detection is characterized by comprising an electrospray ion source, a first group of electrode plates, a second group of electrode plates, a mass spectrometer, a sine wave power supply, a high-field asymmetric waveform power supply, a direct current power supply and a gas generator, wherein one side of the first group of electrode plates is provided with a sample inlet for protein sample ions generated by ionization of the electrospray ion source to enter, the first group of electrode plates and the second group of electrode plates are coaxially and parallelly arranged in front of and behind, a gap is reserved between the first group of electrode plates and the second group of electrode plates, a pre-annealing area is formed between the first group of electrode plates, a separation area is formed between the second group of electrode plates, the rear end of the second group of electrode plates is connected with a detection inlet of the mass spectrometer, and the first group of electrode plates is electrically connected with the sine wave power supply, the outer side of the first group of electrode plates is provided with a temperature control module for controlling the temperature of the pre-annealing area, the second group of electrode plates is electrically connected with the high-field asymmetric waveform power supply and the direct-current power supply, the gas generator is arranged at the front end of the first group of electrode plates, and the generated carrier gas pushes ionized protein sample ions to sequentially pass through the pre-annealing area and the separation area.
2. The device of claim 1, wherein the first and second sets of electrode plates are each formed by two plate-type electrode plates that are symmetrical up and down, and the electrode plates of the first and second sets of electrode plates are spaced at equal intervals.
3. The high-field asymmetric waveform ion mobility device for protein detection according to claim 2, wherein the distance between the electrode plates of the first group of electrode plates and the electrode plates of the second group of electrode plates is 0.1 mm-10 cm, preferably 1 mm-5 mm.
4. The device of claim 1, wherein the sine wave power supply has a voltage amplitude of no less than 3 kV.
5. The device of claim 1, wherein the high-field asymmetric waveform power supply is a square wave, a half sine wave or a double sine wave, and the high-field asymmetric waveform power supply has a voltage amplitude of not less than 3kV and a frequency of 1kHz to 100MHz, preferably 100 kHz.
6. A method of protein detection using the high-field asymmetric waveform ion mobility apparatus of claim 1, comprising the steps of:
firstly, using an electrospray ion source to ionize a target protein sample in a solution to generate protein sample ions, wherein the protein sample ions enter a first group of electrode plates and are pushed by carrier gas to move forwards;
providing a sine wave power supply for the first group of electrode plates, providing a stable temperature field for a pre-annealing area of the first group of electrode plates through a temperature control module, and oscillating the protein sample ions in the pre-annealing area under the combined action of the carrier gas, the sine wave power supply and the temperature field to finish pre-annealing;
thirdly, a high-field asymmetric waveform power supply and a direct-current power supply are provided for the second group of electrode plates, protein sample ions continue to move forward to a separation area in the second group of electrode plates under the pushing of carrier gas, target protein ions are separated in the separation area and enter a mass spectrometer for detection, a mass spectrum signal of scanning compensation voltage is obtained, and the rest ions are neutralized in the separation area;
changing the compensation voltage of the direct current power supply, and repeating the steps to obtain mass spectrum signals under different compensation voltages;
integrating mass spectrum signals under different compensation voltages to obtain a signal spectrogram of the high-field asymmetric waveform ion mobility of the target protein ions.
7. The method of claim 6, wherein the electrospray ion source produces protein sample ions comprising protein positive and negative ions and other ionizable particles.
8. The method for detecting protein by using the high-field asymmetric waveform ion mobility device according to claim 6, wherein the carrier gas is one of hydrogen, helium, nitrogen and sulfur hexafluoride, or a mixture of a plurality of hydrogen, helium, nitrogen and sulfur hexafluoride in any proportion, or a mixture of several of hydrogen, helium, nitrogen and sulfur hexafluoride and a doping gas, the doping gas is selected from ethers, alcohols, ketones and aromatic hydrocarbon gases, and the flow rate of the carrier gas is 0.001L/min to 30L/min, preferably 1L/min to 5L/min.
9. The method for detecting protein by using the high-field asymmetric waveform ion mobility device as claimed in claim 6, wherein the voltage amplitude of the sine wave power supply in the step (II) is not less than 3kV, and the temperature range of the temperature field is 0-500 ℃, preferably 100-300 ℃.
10. The method of claim 6, wherein the compensation voltage of the DC power supply is adjustable between-200V and 200V.
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