CN107179411B - Multi-functional mass spectrum substrate target holder - Google Patents
Multi-functional mass spectrum substrate target holder Download PDFInfo
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- CN107179411B CN107179411B CN201710539749.7A CN201710539749A CN107179411B CN 107179411 B CN107179411 B CN 107179411B CN 201710539749 A CN201710539749 A CN 201710539749A CN 107179411 B CN107179411 B CN 107179411B
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- substrate
- card slot
- target holder
- mass spectrum
- main body
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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 present invention provides a kind of multi-functional mass spectrum substrate target holder, it includes target holder main body and the gene substrate card slot and albumen microorganism substrate card slot that are arranged in the target holder main body, wherein the gene substrate card slot and the albumen microorganism substrate card slot are square structure, and are respectively arranged with card slot through-hole at four angles of the square structure.Wherein the flatness of target holder card slot directly affects Mass Spectrometer Method Mass accuracy, to influence accuracy, this target holder card slot flatness < 10um, Mass Spectrometer Method Mass accuracy < 350ppm greatly improve Mass Spectrometric Identification accuracy.
Description
Technical field
The present invention relates to field of biological detection, more particularly to the multi-functional mass spectrum substrate target holder detected for BIOMARK.
Background technique
Flight time mass spectrum starts from the 1940s, being confined to the backwardness of electronic technology at that time and instrument and equipment, instrument
Resolution ratio is difficult to promote and apply less than 100.Over time with the progress of science and technology, to the end of the eighties,
After Hillenkamp etc. has invented substance assistant laser desorpted ionized new method, by this soft ionization method and flight time
Mass spectrum combination, forms a kind of new instrument: substance assistant laser desorpted ionized/flight time mass spectrum, i.e. MALDI-TOF MS
(Matrix Assisted Laser Desorption Ionization Time of Flight Mass
Spectrometry), it is the novel organic mass spectrometry of a kind of soft ionization developed in recent years, by introducing substrate molecule, is made
Testing molecule does not generate fragment, solves the problems, such as non-volatile and thermal instability large biological molecule desorption ionization, is point
One of the important means of the organic substance of the difficult volatilization of analysis.
The principle of MALDI-TOF MS: when with the laser irradiation sample of some strength and substrate formed cocrystallization film,
Matrix absorbs energy from laser, sample desorption, and electric charge transfer occurs between matrix-sample and makes ionized sample molecule, electricity
From sample accelerate to fly over dirft tube under electric field action, it is different according to the flight time for reaching detector and be detected, root
The ratio between quality charge according to ion (M/Z) is directly proportional to the flight time of ion to detect different ions.
MALDI TOF has become detection and identification polypeptide, protein, polysaccharide, nucleotide, glycoprotein, high polymer in recent years
And the powerful tool of a variety of synthetic polymers, and it is widely used in the drug development of biotechnology and pharmacy corporation, scientific research
Nuclear radiation, chemical substance and monitoring of bio-pathogen of the bioanalysis in field and chemical detection and security department etc..
MALDI-TOF MS has high sensitivity, accuracy height, high resolution, map simplicity, mass range wide and speed
The features such as fast, operationally sample preparation it is easy, can milligram ammonia, extensive, parallelization and increasingly automated processing biological sample to be checked,
And there is special superiority in measurement large biological molecule and synthetic high polymer application aspect.Such as measured using MALDI-TOF MS
The peptide mass fingerprinting of protein digestion cracks (PSD) fragment ion map after composing (PMF), source and combines mass spectrum network data
Library searching can get the sequence of polypeptide, protein.Genome single nucleotide polymorphism (SNPs) is carried out using MALDI-TOF
Analysis detection, can distinguish and identify relative molecular mass up to 7,000 or so (containing more than 20 a bases), there is only 1 base difference
Different DNA.It is worth pointing out that have become must be indispensable in life science proteome research by MALDI-TOF
One of important key technology.
MALDI-TOF can solve in current protein science following several potential challenges, i.e., the discovery of biomarker,
Molecular diagnosis research and development, protein high throughput analysis and proteomic map, but it is not limited to this, MALDI-TOF MS is applied to
The discovery of biomarker and pathogenic mechanism is disclosed on molecular level, to be applied to molecular diagnosis, target is controlled
It treats and personalized medicine, growth, the rule of development and the vital movements such as metabolic regulation and sternly will be more completely prompted for people
The genesis mechanism of weight disease carries out the diagnosis prevention and treatment of disease for the mankind and new drug development provides important theoretical basis.
Traditional MALDI-TOF MS target plate application field is single, such as the MALDI-TOF Axima that SHIMADZU is produced, target
Plate is served only for albumen microbial identification；The MALDI-TOF MASSARRY of AGENA production, target plate are served only for genetic test；Do not have also
There is a kind of covering gene to detect, three kinds of the albumen microbial identification MALDI-TOF mass spectrum target plates applied.
In addition, MALDI-TOF tradition target plate flux is fixed, albumen microorganism substrate flux has 384well, 48well, base
Because substrate flux has 384well, 96well, 24well, cannot a variety of applications selection collocation, and cannot multiple target plates simultaneously into
Sample, multiple laboratory technicians can not test simultaneously, and Mass Spectrometer Method Mass accuracy is low, and identification accuracy is low etc..
Summary of the invention
In order to solve the above-mentioned technical problem, an aspect of of the present present invention provides multi-functional mass spectrum substrate target holder comprising target holder
Main body and the gene substrate card slot being arranged in the target holder main body and albumen microorganism substrate card slot, wherein the gene substrate
Card slot and the albumen microorganism substrate card slot are square structure, and are respectively arranged with card at four angles of the square structure
Slot through-hole.
Preferably, the target holder main body is rectangle, and the direction lead angle as direction signs is provided at one angle,
Lead angle degree is 45 °.
Preferably, multi-functional mass spectrum substrate target holder is further provided with mass spectrum substrate card slot, and outside the target holder main body
Three steps are arranged in the perpendicularly inward direction in surface in the mass spectrum substrate card slot, and the depth of first step is away from the target holder
The outer surface of main body is 0.5mm-1.5mm, and the depth of second step is 1mm-2mm and third platform away from the bottom of the first step
Bottom of the depth of rank away from the second step is 0.3mm-1mm.
Preferably, the flatness < 10um of the first step, Mass Spectrometer Method Mass accuracy < 350ppm.
Preferably, wherein the second step is placed with the magnetic sheet of mass spectrum substrate back.
Preferably, wherein the third step is placed with round sheet magnetic sheet, fixed absorption mass spectrum substrate.
Preferably, wherein the mass spectrum substrate card slot is rectangular, and card is respectively arranged at four rectangular angles
Slot through-hole.
Preferably, multi-functional mass spectrum substrate target holder is further provided with locating groove 2, width 2mm-10mm, height
For 1mm-5mm, positioned for vacuum chamber sample introduction card slot.
Preferably, wherein the quantity of the gene substrate card slot is 4, the quantity of the albumen microorganism substrate card slot is 2,
It can be used for disposable while detecting 4 gene substrates below and/or 2 albumen microorganism substrates below.
Preferably, wherein the material of the target holder selects the group of free metal, silicon wafer and graphene composition.
By the special warfare and superhigh precision of mass spectrum substrate target holder, MALDI-TOF Mass Spectrometer Method Mass accuracy is improved, from
And the accuracy of genetic test, microprotein identification is improved, belong to Mass Spectrometer Method field.
Mass spectrum substrate target holder of the invention is suitable for the CLIN-TOF- of Beijing Yixin Bochuang Biotechnology Co., Ltd.'s production
II clinic flight time mass spectrum.Gene substrate and albumen microorganism substrate can be placed simultaneously, and single injected sampling can carry out simultaneously
Genetic test and albumen microbial identification avoid different detections from distinguishing sample introduction, save the time, single injected sampling can save 15 points
Clock detects 5 times, saves the time 60 minutes within one day, save the time 1320 minutes within one month, a Spring Festival holidays save time 15840 points for one day
Clock.
In addition, mass spectrum substrate target holder flux of the invention is flexible, a target holder can place gene substrate 4 and open, the micro- life of albumen
Object substrate 2 is opened, i.e. 4+2 mode, can also can be arranged according to the detection demand of biological sample with 2+3,6+1 isotype, flux；
Can more people operation of sample preparation simultaneously, a target holder is divided into for+2 albumen microorganism substrates of 4 gene substrates, altogether
It is divided into 6 unit substrates, sample can be prepared simultaneously with 6 laboratory technicians, printing operation, 6 units can be with single injected sampling.
Accuracy is high, and the flatness of target holder card slot directly affects Mass Spectrometer Method Mass accuracy, so that accuracy is influenced, this target
Card slot flatness < 10um, Mass Spectrometer Method Mass accuracy < 350ppm is ask to greatly improve Mass Spectrometric Identification accuracy.
Detailed description of the invention
Fig. 1 mass spectrum substrate target holder floor map.
Fig. 2 mass spectrum substrate target holder diagrammatic cross-section.
Fig. 3 deafness standard items spectrogram.
Fig. 4 microorganism detection Escherichia coli spectrogram.
Fig. 5 genetic test deafness plasmid spectrogram.
Specific embodiment
The existing various exemplary embodiment that the present invention will be described in detail, the detailed description are not considered as to limit of the invention
System, and it is understood as the more detailed description to certain aspects of the invention, characteristic and embodiment.
It should be understood that it is to describe special embodiment that heretofore described term, which is only, it is not intended to limit this hair
It is bright.In addition, for the numberical range in the present invention, it is thus understood that specifically disclose the range upper and lower bound value and they
Between each median.Median and any other statement value in any statement value or stated ranges or in the model
Lesser range is also included in the present invention each of between median in enclosing.These small range of upper and lower bounds can be only
On the spot include or exclude in range.
Unless otherwise stated, all technical and scientific terms used herein has the routine in field of the present invention
The normally understood identical meanings of technical staff.Although the present invention only describes preferred method and material, of the invention
Implement or also can be used and similar or equivalent any method and material described herein in testing.
Embodiment one, target holder flatness influence qualification result accuracy
By taking deaf gene standard items as an example, totally 18 mass spectra peaks, mass range 3000-9000Da, select three kinds of flatnesses
10 μm, 20 μm, 30 μm, the same sample of every kind of flatness does 12 repetitions, analyzes the mass deviation at every peak of contrasting detection result,
Calculate mass accuracy.
Parameter setting:
Turing mode:linear
Mass Range:3000-9000
Max Laser Rep Rate:10.0
Power:90
Profiles:40
Shots:10
1) 10 μm of substrate slot bit plane degree <, 12 repetitions, each site peak deviation: (grey is maximum deviation)
Maximum deviation is less than 350ppm in the position 8214.4Da, deviation 2.78Da, mass accuracy.
2) 20 μm of substrate slot bit plane degree <, 12 repetitions, each site peak deviation: (grey is maximum deviation)
Maximum deviation is less than 500ppm in the position 8214.4Da, deviation 3.89Da, mass accuracy.
3) 30 μm of substrate slot bit plane degree <, 12 repetitions, each site peak deviation: (grey is maximum deviation)
Maximum deviation is less than 550ppm in the position 8214.4Da, deviation 4.26Da, mass accuracy.
Fig. 3 deafness standard items spectrogram is seen referring specifically to spectrogram.Target holder flatness is to spectrum it can be seen from Fig. 3 and upper table
Plot quality accuracy is affected, and the index that the present invention reaches is: 10 μm of target holder flatness <, mass accuracy < 350ppm.
Embodiment two, microbial identification
5 kinds of fresh bacterial strains of microorganism, 3 repetitions of every kind of bacterial strain are selected, mass range 2000-20000Da acquires data and exists
The marking of BE3.0 software, checks confidence level result.
Parameter setting:
Turing mode:linear
Mass Range:2000-20000
Max Laser Rep Rate:30.0
Power:60
Profiles:100
Shots:5
Spectrogram is shown in Fig. 4 microorganism detection Escherichia coli spectrogram.
Conclusion: confidence level > 25 divides, and is that qualification result is credible, this 15 groups of data authentication result of experiment is correct, correctly
Rate 100%.
Embodiment three, genetic test
Deaf plasmid sample is selected, totally 20 heavy spectral peak, mass range 3000-9000Da, 5 repetitions of positive sample, yin
Property 5 repetitions of sample, analysis detection result identify accuracy.
Parameter setting:
Turing mode:linear
Mass Range:3000-9000
Max Laser Rep Rate:10.0
Power:90
Profiles:40
Shots:10
Deaf plasmid sample qualification result:
Spectrogram is shown in Fig. 5 genetic test deafness plasmid spectrogram.
Conclusion: based on 10 μm of target holder of flatness <, 10 deaf plasmid samples, 20 positions of each deafness plasmid are detected
Point, totally 200 sites, qualification result accuracy rate 100%.
The basic principles, main features and advantages of the present invention have been shown and described above.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, the present invention
Claimed range is delineated by the appended claims, the specification and equivalents thereof from the appended claims.
Claims (3)
1. a kind of multi-functional mass spectrum substrate target holder comprising target holder main body and the gene substrate card being arranged in the target holder main body
Slot (3) and albumen microorganism substrate card slot (4), wherein
The target holder main body is rectangle, which is selected from by silicon wafer or graphene, wherein in one jiao of side of being arranged as of main body
To the oblique angle lead angle (1) of mark, lead angle degree is 45 °, and is respectively arranged in the top and bottom of main body and extends to main body or so two
The locating groove (2) at end is positioned for vacuum chamber sample introduction card slot；
The gene substrate card slot (3) is arranged in parallel in main body left part, and right part is equipped with multiple albumen microorganism substrate card slots in parallel,
Wherein mass spectrum substrate card slot is rectangular, and is respectively arranged with card slot through-hole (8) at four angles of the rectangular card slot, so as to
Clamp substrate；
The substrate card slot direction perpendicularly inward along the target holder body outer surface, successively overlaps three steps:
First step (5) flatness < 10um, Mass Spectrometer Method Mass accuracy < 350ppm, and its depth is away from the target holder master
The outer surface of body is 0.5mm-1.5mm, for placing substrate；
Bottom of the depth away from the first step of second step (6) is 1mm-2mm, for placing the magnetic of mass spectrum substrate back
Piece；
Bottom of the depth away from the second step of third step (7) is 0.3mm-1mm, is placed with round sheet magnetic sheet, is used for
Fixed absorption mass spectrum substrate.
2. the multi-functional mass spectrum substrate target holder according to claim 1, wherein the width of locating groove (2) is 2mm-10mm,
Height is 1mm-5mm.
3. according to claim 1 or multi-functional mass spectrum substrate target holder described in 2, wherein the quantity of the gene substrate card slot is
4, the quantity of the albumen microorganism substrate card slot is 2, can be used for 4 gene substrates below of disposable detection simultaneously with/
Or 2 albumen microorganism substrates below.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201710539749.7A CN107179411B (en) | 2017-07-05 | 2017-07-05 | Multi-functional mass spectrum substrate target holder |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710539749.7A CN107179411B (en) | 2017-07-05 | 2017-07-05 | Multi-functional mass spectrum substrate target holder |
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| CN107179411A CN107179411A (en) | 2017-09-19 |
| CN107179411B true CN107179411B (en) | 2019-01-29 |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108267500A (en) * | 2018-01-03 | 2018-07-10 | 北京毅新博创生物科技有限公司 | Mass spectrum substrate target holder detects the purposes of biological sample for BIOMARK |
| CN108148742A (en) * | 2018-01-03 | 2018-06-12 | 北京毅新博创生物科技有限公司 | A kind of mass spectrum substrate target holder for detection of biological samples |
| CN108709927B (en) * | 2018-05-24 | 2021-09-07 | 华东理工大学 | Disposable mass spectrum target plate and preparation method and application thereof |
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| US7297942B2 (en) * | 2003-01-15 | 2007-11-20 | Bruker Daltonik, Gmbh | Method and device for cleaning desorption ion sources |
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| WO1999000657A1 (en) * | 1997-06-26 | 1999-01-07 | Perseptive Biosystems, Inc. | High density sample holder for analysis of biological samples |
| US6825478B1 (en) * | 2003-10-10 | 2004-11-30 | Perseptive Biosystems, Inc. | MALDI plate with removable magnetic insert |
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