Method and device for the qualitative and/or quantitative determination of a protein and/or peptide pattern of a fluid sample, which has been taken from a human or animal body
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Publication Number
US-7955862-B2
Publication Date
2011-06-07
Expiration Date
Abstract
Provided are a process and a device for the qualitative and/or quantitative determination of a protein and/or peptide pattern of a liquid sample taken from a human or animal body for checking its state. The peptides and proteins of the liquid sample are processed and then subjected to analysis, wherein reference and sample values describing states of a human or animal body as well as deviations and correspondences derived therefrom are established, automatically stored in a data base, and when the protein and/or peptide pattern is again determined, a search for optimum correspondence is automatically performed. In this process, mass components or structural components of the proteins and/or peptides are established and stored. Subsequently, real masses or real structures are calculated from a common evaluation of the stored mass components or structural components, and an assignment to the proteins and/or peptides contained in the liquid sample is performed from a combination of the calculated real masses or real structures.
Core Innovation
The disclosed invention relates to a process for the determination of a protein and/or peptide pattern of a liquid sample taken from a human or animal body. The process separates the liquid sample into fractions with a provided separation time for each fraction and analyzes the fractions with a mass spectrometer providing masses and amplitudes, and a three-dimensional spectrum of the liquid sample is generated such that the first dimension represents the amplitude of the spectral lines, the second dimension represents separation time, and the third dimension represents the mass.
The three-dimensional spectrum is evaluated to yield real separation time, real masses, and related overall amplitudes of the proteins and/or peptides of the liquid sample. The resulting real separation time, real masses, and related overall amplitudes are defined as a protein and/or peptide pattern of the sample, and the system/process includes automated determination using mass spectrometry with separation into fractions and subsequent generation and evaluation of the three-dimensional spectral representation.
The description further states that mass/structural components are stored to form a multidimensional (3D) spectral data field and that real masses and overall amplitudes are computed via algorithms after interference elimination. Proteins/peptides are assigned by comparing calculated real masses and structures to stored reference patterns, and calibration is supported by using proteins/peptides present in all samples with approximately constant concentration.
Claims Coverage
The independent claim covers an end-to-end process that fractionates a human/animal body liquid sample, analyzes fractions by mass spectrometry to obtain masses and amplitudes, generates a three-dimensional spectrum, and evaluates it to yield a protein and/or peptide pattern. Four inventive features are identified across the independent claim and dependent refinements.
Three-dimensional protein/peptide pattern from fraction-separated mass spectrometric analysis
Separating the liquid sample into fractions with separation time, analyzing the fractions with a mass spectrometer providing masses and amplitudes, generating a three-dimensional spectrum with amplitude, separation time, and mass dimensions, and evaluating the three-dimensional spectrum to yield real separation time, real masses, and related overall amplitudes as a protein and/or peptide pattern.
Capillary electrophoresis with direct ionization and on-line coupled mass spectrometer detection
Obtaining the three-dimensional spectrum by separation via capillary electrophoresis, applying direct ionization, interfacing the separated results to an on-line coupled mass spectrometer, and detecting the result.
Interference elimination before evaluation of stored spectra
Eliminating interferences in a three-dimensional spectrum after storing the three-dimensional spectra and before evaluating the three-dimensional spectra.
Evaluating only masses and overall amplitudes above preset thresholds
Evaluating only masses and overall amplitudes that are above preset threshold values.
Across the independent claim and its dependent refinements, the inventive concept is generating and evaluating a three-dimensional spectrum from fraction-separated mass spectrometric analysis to obtain real separation time, real masses, and overall amplitudes as a protein/peptide pattern, with additional specifications regarding capillary electrophoresis/direct ionization/on-line MS detection, interference elimination, and threshold-limited evaluation.
Stated Advantages
Produces a protein and/or peptide pattern by evaluating the three-dimensional spectrum to yield real separation time, real masses, and related overall amplitudes.
Supports assignment of proteins/peptides by comparing calculated real masses/structures to stored reference patterns.
Computes real masses and overall amplitudes using algorithms after interference elimination.
Enables calibration of masses and overall amplitudes using proteins/peptides present in all samples with approximately constant concentration.
Documented Applications
Distinguishing renal diseases using urine, with marker polypeptides and typical protein/peptide spectral patterns, including comparisons among MNGN, MCG, and FSGS groups versus healthy groups.
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