CN104359967B - A kind of analytic method of the overlapping isotope profile of biological mass spectrometry - Google Patents
A kind of analytic method of the overlapping isotope profile of biological mass spectrometry Download PDFInfo
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Abstract
The present invention relates to a kind of analytic method of the overlapping isotope profile of biological mass spectrometry.This analytic method is based on the mass spectrographic original firsts and seconds mass spectrum, by calculate overlapping isotopic peak gedanken experiment intensity and its and actual experiment intensity deviation so as to calculating actual distribution intensity of the overlapping isotopic peak in each isotope profile.Compared with prior art, parsing and fractionation amount of calculation of the analytic method of the present invention to the intensity of overlapping isotopic peak are small, and flux is high, and the degree of accuracy is high, suitable for large biological molecule (such as protein, carbohydrate) mass spectrum and tandem mass spectrometry efficient parsing and structure precise Identification.
Description
Technical field
The present invention relates to a kind of analytic method of the overlapping isotope profile of biological mass spectrometry, relate generally to related to biological mass spectrometry
System biology field, including proteomics, metabolism group, sugar group learn etc. technical field.
Background technology
Development and business with Soft ionization techniques (such as electron spray ionisation) and High Resolntion Mass-Spectrometry analyzer (such as orbit trap)
Change, mass spectrum is increasingly being used for the analysis of large biological molecule (such as protein, polysaccharide).
The A of Chinese patent CN 103389335 disclose one kind and identify biology by analyzing large biological molecule mass spectrometric data
The analytical equipment and method of macromolecular primary structure and composition.The analytical equipment and method are based on the original of the large biological molecule
Firsts and seconds mass spectrum, by isotopic peak mass-to-charge ratio and profile fingerprint comparison so as to identify the large biological molecule.On
The Initial experiments mass spectrometric data that analytical equipment and method are directly gathered using mass spectrograph is stated, it is same to precursor ion and fragment ion
The accurate mass-to-charge ratio and relative intensity of each Initial experiments isotopic peak are compared with corresponding theoretical value in the plain profile in position, point
Candidate large biological molecule Yong Yu not be found from database and confirms wherein confidence level highest one using tandem mass spectrometry, so as to
The qualitative and quantitative analysis of high confidence level is carried out to biomolecule.
The charged ion (precursor ion, first mass spectrometric) of these macromoleculars of mass-spectrometer measurement and corresponding fragment (fragment ion,
Second order mses).Mass spectrum is the two-dimentional spectrogram that transverse axis is mass-to-charge ratio, the longitudinal axis is intensity.Due to the presence of isotope, all ions
(including precursor ion and fragment ion) is the isotope profile comprising several isotopic peaks in mass spectrum.Because mass spectrum is horizontal
Axle, that is, analysis of the mass analyzer to mass-to-charge ratio have certain scope (such as 500-2000), and large biological molecule is broken
Piece is more (and each fragment typically occurs with multiple valence states), there is more intensive the isotope profile of fragment ion in second order mses
It is overlapping.Method of the tradition based on " deconvoluting " (also referred to as removing isotope) fails to carry out effectively these overlapping isotope profiles
Parsing and fractionation；" the isotope profile fingerprint comparison " of newly-developed although the ratio in algorithm is high with the point-score degree of accuracy,
Speed is slower.
The content of the invention
It is an object of the present invention to overcome the above-mentioned drawbacks of the prior art and provide a kind of amount of calculation is small, flux
The analytic method of the overlapping isotope profile of biological mass spectrometry high, the degree of accuracy is high.
The purpose of the present invention can be achieved through the following technical solutions：
A kind of analytic method of the overlapping isotope profile of biological mass spectrometry, comprises the following steps：
(1) found out by experiment isotope profile in mass spectrogram with the fingerprint comparison of theoretical isotope profile in database overlapping
The theoretical relative intensity of ion and its all isotopic peaks and experiment absolute intensity, including overlapping isotopic peak are theoretical relatively strong
Degree and actual experiment absolute intensity sum；
(2) top non-overlapping using in each theoretical isotope profile for participating in overlapping ion is as reference peak, to phase
The laboratory strength of each isotopic peak is normalized in the experiment isotope profile answered；
(3) gedanken experiment absolute intensity of the overlapping isotopic peak in each overlapping ion is calculated；
(4) overlapping isotopic peak actual experiment absolute intensity sum is calculated relative to the strong of gedanken experiment absolute intensity sum
Spend deviation；
(5) actual experiment absolute intensity of the overlapping isotopic peak in the overlapping ion of each participation is calculated.
In step (3), gedanken experiment absolute intensity of the overlapping isotopic peak in each overlapping ion passes through below equation
It is calculated：
The theoretical relative intensity * of the overlapping isotopic peak of the gedanken experiment absolute intensity of each overlapping isotopic peak=respectively participate in
With reference to isotopic peak actual experiment absolute intensity/refer to isotopic peak theory relative intensity.
In step (4), overlapping isotopic peak actual experiment absolute intensity sum is relative to gedanken experiment absolute intensity sum
Strength variance be calculated by below equation：
Overlapping isotopic peak actual experiment absolute intensity sum relative to gedanken experiment absolute intensity sum strength variance
=[the actual experiment absolute intensity sum of overlapping isotopic peak-(gedanken experiment of the overlapping isotopic peak in each overlapping ion is exhausted
To intensity summation)]/(gedanken experiment absolute intensity summation of the overlapping isotopic peak in each overlapping ion).
In step (5), actual experiment absolute intensity of the overlapping isotopic peak in the overlapping ion of each participation passes through following
Formula is calculated：
Actual experiment absolute intensity=overlapping isotopic peak of the overlapping isotopic peak in the overlapping ion of each participation is each
(the overlapping isotopic peak actual experiment absolute intensity sums of 1+ are relative to preferable real by gedanken experiment absolute intensity * in overlapping ion
Test the strength variance of absolute intensity sum).
According to actual experiment absolute intensity of the overlapping isotopic peak being calculated in the overlapping ion of each participation, make
Isotope profile fingerprint comparison figure after parsing.
Method of the present invention is equally applicable to overlapping profile information sum in other mass spectrums and any 2-D data
According to parsing.
Compared with prior art, analytic method of the invention is based on the mass spectrographic original firsts and seconds mass spectrum, passes through
Calculate the gedanken experiment intensity of overlapping isotopic peak and its exist with the deviation of actual experiment intensity so as to calculate overlapping isotopic peak
Actual distribution intensity in each overlapping isotope profile.Parsing of the analytic method of the present invention to the intensity of overlapping isotopic peak and
It is small to split amount of calculation, flux is high, and the degree of accuracy is high, suitable for large biological molecule (such as protein, carbohydrate) tandem mass spectrometry efficient parsing
And structure precise Identification.
Brief description of the drawings
Figure 1A is that fragment ion y52-6+ mass-to-charge ratioes are in ubiquitin protein energetic encounter induction dissociation second order mses
Isotope profile fingerprint ratio before the 982.37582 and 983.388428 overlapping parsing of overlapping isotopic peak (arrow marks in figure)
To figure；
Figure 1B is that fragment ion y52-6+ mass-to-charge ratioes are in ubiquitin protein energetic encounter induction dissociation second order mses
Isotope profile fingerprint ratio after the 982.37582 and 983.388428 overlapping parsing of overlapping isotopic peak (arrow marks in figure)
To figure；
Fig. 2A is fragment ion y61-H in ubiquitin protein energetic encounter induction dissociation second order mses2O-7+ mass-to-charge ratioes are
Isotope profile fingerprint ratio before the 982.37582 and 983.388428 overlapping parsing of overlapping isotopic peak (arrow marks in figure)
To figure；
Fig. 2 B are fragment ion y61-H in ubiquitin protein energetic encounter induction dissociation second order mses2O-7+ mass-to-charge ratioes are
Isotope profile fingerprint ratio after the 982.37582 and 983.388428 overlapping parsing of overlapping isotopic peak (arrow marks in figure)
To figure.
M/z is mass-to-charge ratio in figure, and Abundance (a.u.) is the absolute laboratory strength of arbitrary unit；Theo. it is theoretical same position
Plain profile, Exp. are experiment isotope profile.
Embodiment
The present invention is described in detail with specific embodiment below in conjunction with the accompanying drawings.
Embodiment
A kind of analytic method of the overlapping isotope profile of biological mass spectrometry, comprises the following steps：
(1) found out by experiment isotope profile in mass spectrogram with the fingerprint comparison of theoretical isotope profile in database overlapping
The theoretical relative intensity of ion and its all isotopic peaks and experiment absolute intensity, including overlapping isotopic peak are theoretical relatively strong
Degree and actual experiment absolute intensity sum；
(2) top non-overlapping using in each theoretical isotope profile for participating in overlapping ion is as reference peak, to phase
The laboratory strength of each isotopic peak is normalized in the experiment isotope profile answered；
(3) gedanken experiment absolute intensity of the overlapping isotopic peak in each overlapping ion is calculated：
The theoretical relative intensity * of the overlapping isotopic peak of the gedanken experiment absolute intensity of each overlapping isotopic peak=respectively participate in
With reference to isotopic peak actual experiment absolute intensity/refer to isotopic peak theory relative intensity；
(4) overlapping isotopic peak actual experiment absolute intensity sum is calculated relative to the strong of gedanken experiment absolute intensity sum
Spend deviation：
Overlapping isotopic peak actual experiment absolute intensity sum relative to gedanken experiment absolute intensity sum strength variance
=[the actual experiment absolute intensity sum of overlapping isotopic peak-(gedanken experiment of the overlapping isotopic peak in each overlapping ion is exhausted
To intensity summation)]/(gedanken experiment absolute intensity summation of the overlapping isotopic peak in each overlapping ion)；
(5) actual experiment absolute intensity of the overlapping isotopic peak in the overlapping ion of each participation is calculated：
Actual experiment absolute intensity=overlapping isotopic peak of the overlapping isotopic peak in the overlapping ion of each participation is each
(the overlapping isotopic peak actual experiment absolute intensity sums of 1+ are relative to preferable real by gedanken experiment absolute intensity * in overlapping ion
Test the strength variance of absolute intensity sum).
Then the actual experiment absolute intensity according to the overlapping isotopic peak being calculated in the overlapping ion of each participation,
Make the isotope profile fingerprint comparison figure after parsing.
Y52-6+ and y61-H in dissociation second order mses is induced with ubiquitin protein energetic encounter below2O-7+ two it is overlapping from
Illustrate the specific implementation step and result of this analytic method exemplified by the parsing and fractionation of son, as shown in table 1 and Figure 1A~Fig. 2 B.Table
1 row and column is marked with Arabic numerals and English alphabet respectively, and grid is then represented with the combination of letter and number, such as
B6 represents the row of column b the 6th.For overlapping isotopic peak of the mass-to-charge ratio at 982.385132, the first step uses B7*D8/B8 respectively
Y52-6+ and y61-H is calculated with B12*D13/B132Number in the gedanken experiment absolute intensity of O-7+ ions, i.e. grid E3 and F3
Value；Second step, it is exhausted relative to gedanken experiment to calculate actual experiment absolute intensity sum with formula (D3- (E3+F3))/(E3+F3)
To the strength variance of intensity sum, i.e. numerical value in grid G3；3rd step, counted respectively with formula E3* (1+G3) and F3* (1+G3)
Calculate y52-6+ and y61-H2O-7+ ions mass-to-charge ratio is the actual experiment absolute intensity of 982.385132 isotopic peak.From table 1
Arrange H and arrange in G from the point of view of the comparison of relative intensity deviation, the error after parsing have dropped 88 and 125 percentages respectively before relatively parsing
Point.Y52-6+ and y61-H2Another overlapping isotopic peak (mass-to-charge ratio 983.388428) of O-7+ ions is by above-mentioned same
Result after step is parsed and split is also shown in Table 1 below.The front and rear isotope of the overlapping isotopic peak parsing of y52-6+ ions
Profile fingerprint comparison figure is respectively at shown in Figure 1A, Figure 1B.y61-H2The front and rear isotope of the overlapping isotopic peak parsing of O-7+ ions
Profile fingerprint comparison figure is as shown in Fig. 2A, Fig. 2 B.
Table 1
Table 1 only lists y52-6+ and y61-H2The overlapping isotopic peak of two ions of O-7+ and for test definitely it is strong
Spend the isotopic peak for being normalized and (being transformed into relative intensity).It is inclined to test relative intensity, mass-to-charge ratio deviation and relative intensity
The calculation formula of difference is as follows as 982.37582 isotopic peak demonstration using mass-to-charge ratio：Experiment relative intensity (E7)=D7*B8/D8,
Mass-to-charge ratio deviation (F7)=(C7-A7) * (1E-6)/A7, relative intensity deviation (G7)=E7-B7.
Relative intensity deviation in table 1 in H6 and H11 is the deviation before overlapping parsing and fractionation.
The above-mentioned description to embodiment is understood that for ease of those skilled in the art and using invention.
Person skilled in the art obviously can easily make various modifications to these embodiments, and described herein general
Principle is applied in other embodiment without by performing creative labour.Therefore, the invention is not restricted to above-described embodiment, ability
Field technique personnel do not depart from improvement that scope made and modification all should be the present invention's according to the announcement of the present invention
Within protection domain.
Claims (2)
1. the analytic method of the overlapping isotope profile of a kind of biological mass spectrometry, it is characterised in that comprise the following steps：
(1) overlapping ion is found out with the fingerprint comparison of theoretical isotope profile in database by testing isotope profile in mass spectrogram
And its theoretical relative intensity of all isotopic peaks with experiment absolute intensity, including the theoretical relative intensity of overlapping isotopic peak with
Actual experiment absolute intensity sum；
(2) top non-overlapping using in each theoretical isotope profile for participating in overlapping ion is as reference peak, to corresponding
The laboratory strength of each isotopic peak is normalized in experiment isotope profile；
(3) gedanken experiment absolute intensity of the overlapping isotopic peak in each overlapping ion is calculated；
(4) it is inclined relative to the intensity of gedanken experiment absolute intensity sum to calculate overlapping isotopic peak actual experiment absolute intensity sum
Difference；
(5) actual experiment absolute intensity of the overlapping isotopic peak in the overlapping ion of each participation is calculated；
In step (3), gedanken experiment absolute intensity of the overlapping isotopic peak in each overlapping ion is calculated by below equation
Obtain：
The theoretical relative intensity * references of the overlapping isotopic peak of the gedanken experiment absolute intensity of each overlapping isotopic peak=respectively participate in
Isotopic peak actual experiment absolute intensity/refer to isotopic peak theory relative intensity；
In step (4), overlapping isotopic peak actual experiment absolute intensity sum is strong relative to gedanken experiment absolute intensity sum
Degree deviation is calculated by below equation：
Overlapping isotopic peak actual experiment absolute intensity sum relative to gedanken experiment absolute intensity sum strength variance=[weight
The actual experiment absolute intensity sum of folded isotopic peak-(gedanken experiment of the overlapping isotopic peak in each overlapping ion is definitely strong
Spend summation)]/(gedanken experiment absolute intensity summation of the overlapping isotopic peak in each overlapping ion)；
In step (5), actual experiment absolute intensity of the overlapping isotopic peak in the overlapping ion of each participation passes through below equation
It is calculated：
Actual experiment absolute intensity=overlapping isotopic peak of the overlapping isotopic peak in the overlapping ion of each participation is each overlapping
(the overlapping isotopic peak actual experiment absolute intensity sums of 1+ are exhausted relative to gedanken experiment by gedanken experiment absolute intensity * in ion
To the strength variance of intensity sum).
A kind of 2. analytic method of the overlapping isotope profile of biological mass spectrometry according to claim 1, it is characterised in that according to
Actual experiment absolute intensity of the overlapping isotopic peak being calculated in the overlapping ion of each participation, makes the same position after parsing
Plain profile fingerprint comparison figure.
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| CN111551626A (en) * | 2020-05-18 | 2020-08-18 | 苏州市汉诺生物科技有限公司 | Cascade mass spectrometry analysis method based on molecular composition and structural fingerprint identification |
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| CN101115991A (en) * | 2005-05-13 | 2008-01-30 | 株式会社岛津制作所 | Mass analysis data analysis device and program |
| CN101384898A (en) * | 2006-06-08 | 2009-03-11 | 株式会社岛津制作所 | Data processing device for chromatograph mass spectrum analysis |
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| JP2001160373A (en) * | 1999-12-02 | 2001-06-12 | Hitachi Ltd | Ion trap mass spectrometry and ion trap mass spectrometer |
| WO2006106724A1 (en) * | 2005-03-31 | 2006-10-12 | National Institute Of Advanced Industrial Science And Technology | Method of protein analysis, apparatus and program |
| JP4821400B2 (en) * | 2006-03-28 | 2011-11-24 | 株式会社島津製作所 | Structural analysis system |
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| CN101115991A (en) * | 2005-05-13 | 2008-01-30 | 株式会社岛津制作所 | Mass analysis data analysis device and program |
| CN101384898A (en) * | 2006-06-08 | 2009-03-11 | 株式会社岛津制作所 | Data processing device for chromatograph mass spectrum analysis |
| CN101871945A (en) * | 2010-06-13 | 2010-10-27 | 中国科学院计算技术研究所 | Spectrum library generating method and spectrogram identifying method of tandem mass spectrometry |
| CN103389335A (en) * | 2012-05-11 | 2013-11-13 | 中国科学院大连化学物理研究所 | Analysis device and method for identifying biomacromolecules |
| CN103245714A (en) * | 2013-03-25 | 2013-08-14 | 暨南大学 | Protein secondary mass spectrum identification method of marker loci based on candidate peptide fragment discrimination |
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