MXPA06002442A - Device and method for the quantitative evaluation of the polypeptides contained in a body fluid sample, and marker for the detection of pathological states - Google Patents

Abstract

Disclosed are devices and methods for quantitatively evaluating the polypeptides contained in a body fluid sample and markers for detecting pathological states. The polypeptides are compared to reference values stored in a database. Said reference values are stored as data records of state-relevant polypeptides, which comprise at least one respective indication about the probability for the polypeptides to occur or the concentration thereof for a pathological state in samples of healthy and ill subjects.

Description

DEVICE AND METHOD FOR THE QUANTITATIVE EVALUATION OF THE POLYPEPTIDES CONTAINED IN A SAMPLE OF BODY FLUID, AND MARKER TO DETECT PATHOLOGICAL STATES DESCRIPTION OF THE INVENTION The invention relates to devices and methods for the quantitative evaluation of the polypeptides contained in a body fluid sample and the comparison with reference values stored in a data bank as well as markers to recognize disease states. A method and a device for the qualitative and / or quantitative determination of a protein and / or polypeptide standard from a fluid sample is known from DE 100 21 737 C2. The proteins and / or the polypeptides of a fluid sample are separated by capillary electrophoresis, then directly ionized and transferred online through an interface to a mass spectrometer coupled thereto. To control the state of a human or animal body over a longer period of time, the reference and test values that describe states as well as the deviations and coincidences that result from this are automatically stored in a data bank. a new determination of a protein and / or polypeptide pattern automatically searches for the best possible matches. REF.:170518 It has been proven that with the polypeptide patterns registered up to now it is already possible to make very reliable statements about the states of the human or animal body. These can complement or substitute the usual research and diagnostic methods. In particular, for diabetic and diabetic nephropathy, numerous reference and test values have been obtained for the determination of polypeptide patterns. Diabetes is often a previous stage of diabetic clothing that can develop over years and decades. Nephropathy goes through several states in diabetes. With the currently available means it is hardly possible to make a premature diagnosis of diabetic nephropathy, and only at great expense and also only at a relatively late time. The timely diagnosis and consequent therapy of manifest nephropathy would not only prevent or delay the need for dialysis, but also reduce the high cardiovascular risk of the patient with diabetes. It has been found that the polypeptide patterns of a fluid sample, for example from urine, allow a diagnosis in a premature state of the disease. Other investigations allow us to recognize that by means of polypeptide patterns it is also possible to diagnose other diseases different from diabetes and diabetic nephropathy mentioned above. The invention aims to indicate a definition of the polypeptides suitable for computer-controlled storage and evaluation, useful for a device and a method for the quantitative evaluation of the polypeptides contained in a body fluid sample and its comparison with the values of reference stored in a data bank. This problem is solved in a device for the quantitative evaluation of the polypeptides contained in a body fluid sample and its comparison with the reference values stored in a database characterized in that the reference values are stored as data registers of State-relevant polypeptides which in each case comprise at least one indication on the probability of the occurrence and / or concentration of polypeptides for a disease state in samples from healthy and diseased subjects, and in a method for the quantitative evaluation of the contained polypeptides in a body fluid sample and its comparison with reference values stored in a data bank that is characterized because reference values are used as records for comparison of polypeptide data relevant to the state by comparing an indication on the probability of the occurrence and / or concentration of the polypeptides in the body fluid sample with the indication of in each case at least one reference value on the probability of occurrence and / or concentration of the polypeptides for a pathological state in samples from healthy and sick subjects. The invention further has the object of specifying a marker for recognizing disease states by a suitable definition for the storage and evaluation of the contained polypeptides. This problem is solved with a marker to recognize pathological states characterized by a multitude of state-relevant polypeptides which in each case are linked with an indication about the probability of the occurrence and / or concentration of the polypeptide for a pathological state in samples from healthy subjects and sick. The improvements and favorable configurations are derived from the respective subordinate claims. With the invention, the polypeptide data whose concentration varies markedly in the body fluid in the case of a pathological condition compared to a normal state are evaluated. These polypeptides can be determined in previous studies with a multitude of test subjects.

Each polypeptide is linked to indications that for a pathological condition comprise an indication about the probability of occurrence or concentration in a healthy subject and in a sick subject. With the comparison of the polypeptides of fluid samples with reference values, it results for each polypeptide of the fluid sample compared an association in the sense that if its occurrence and / or concentration represents the healthy state or the diseased state. By comparing a multitude of polypeptides it is possible to reduce or prevent counterfeiting of the overall result due to isolated individual deviations from the probability of occurrence and typical concentrations. In accordance with an improvement it is possible to define the polypeptides by indicating their relevant mass and their relevant retention time in capillary electrophoresis. In this way, the data of capillary electrophoresis and mass spectrometry are directly adopted. These specifications are different but unequivocal for all polypeptides, so that the specifications are sufficient for the definition. The retention time in capillary electrophoresis can be determined, for example, by capillary electrophoresis with the use of a 90 cm long glass capillary with an internal diameter (ID) of 75 μm and an external diameter (OD) of 360 μm with a tension of 30 kV, being that 30% of methanol, 0.5% of formic acid in water are used as solvent for the sample. For the diagnosis of "diabetes" and "kidney injury", special data records of state-relevant polypeptides were compiled into a data bank, which were determined and verified by urine samples from a multitude of test subjects. It is possible to compare here individual polypeptides, a combination of polypeptides or all polypeptides. In view of the fact that the representativeness of many polypeptides is redundant for the healthy state and the pathological state, in the simplest case it could be sufficient with the comparison of only one of these polypeptides. However, in order to eliminate errors due to statistical insecurities or individual deviations, one must aspire to compare a combination of state-relevant polypeptides. An optimal result is obtained if all the listed polypeptides are compared. These polypeptides that serve as markers can potentially also be therapeutic targets. Thus, the development of therapeutic agents that either rely on these polypeptides or have them as an objective structure is possible. In this, the occurrence and / or concentration of the polypeptides in each case is modified in the body by complementation and / or antibodies, so that their concentration returns to normal values in the body fluid examined. The invention is explained below by means of the figures. They show: Figure la-le graphical representations of individual intermediate stages in obtaining data for the definition of polypeptide patterns. Figure 2a- 2f graphic representations of polypeptide patterns and their relation. In an exemplary embodiment of the invention, the presence and concentration of a large number of polypeptides in urine is analyzed. This happens now by means of capillary electrophoresis coupled to a mass spectrometer (CE-MS), but nevertheless it can also be carried out objectively with other methods. The results of the CE-MS measurement symbolized by the graph in the figure lead, after the three-dimensional representation by application of the retention time on the x-axis, the mass on the y-axis, and the signal amplitude on the x-axis. a polypeptide pattern of typically 500-1500 polypeptides. Figure lb shows the figure of the "raw data" from which the relevant signals are first separated by filtering, in the figure they are designated as "legitimate signals" by means of a software (software) for filtering and evaluation, and then a polypeptide pattern, designated in Figure Id as a "polypeptide standard, is calculated.The polypeptides are scored / identified by their mass and retention time in the EC.Their concentration is calculated by the amplitude of the signal. The data that constitute the polypeptide pattern are deposited in a data bank In a separate data set, the information necessary for the unambiguous identification is stored for each relevant peptide The figure symbolically shows a representation on the screen of several data records. In order to recognize the essential polypeptides, the urine of patients with and without diagnosed diabetic nephropathy and respectively with and without diagnosed diabetes was examined by CE-MS, by comparison in the database of more than 50 measurements it was possible to elaborate a typical polypeptide pattern subjugated with healthy kidney. With the same technology, urine samples from more than 200 patients with type I and type II diabetes were measured. These patients constitute groups of different states of renal diseases, from completely imperceptible to values higher than 3 g protein / day in the urine. For the analysis of the data, the patients were divided into four groups, as shown in the following table.

A specific peptide pattern is developed from the pooled polypeptide samples of an examined group. The polypeptide standards obtained in this manner show typical deviations compared to normal samples, ie, changes in individual polypeptides. For clarification, the graphic representations of the polypeptide patterns specific to the group according to the table are shown in figure 2.

The polypeptide patterns of the group KO, PO, Pl, P2 are reproduced in each case in figures 2a, 2b, 2c, 2d. From the polypeptide patterns of the groups KO, PO, Pl, P2 a synthetic global concept is compiled, and this is used to elaborate a polypeptide pattern according to figure 2a as a marker profile. For the search of relevant markers only polypeptides were used that could be found in at least one of the groups in a relevant amount, in this case greater than 40%. Next, those polypeptides that allow differentiation between the "healthy" and "sick" classes were considered. In this aspect, a consistent development on the groups was taken into account. Variations in the polypeptide pattern are partly conditioned by the disease causing diabetes, and therefore can be detected uniformly in all diabetics, partly conditioned by or also cause of incipient / progressive nephropathy. Therefore, it is possible to resort to these polypeptides as markers for the diagnosis of diabetes and diabetic nephropathy. The polypeptides present there are then searched in the samples of patients being examined, after which their presence or deficiency is used for diagnosis, as shown in Figure 2f. The following table compiles the particularly relevant marker polypeptides found within the context of the detection of diabetic nephropathy. The polypeptides mentioned herein serve to detect the disease in an incipient state, and for this they can be used individually, in extracts or in fully combined. The list contains 380 polypeptides defined by their mass and their retention time in capillary electrophoresis. The consecutive numbers 1 to 157 constitute the marker peptides used for the diagnosis "diabetes". Consecutive numbers 158 to 380 comprise the marker peptides which are used for the diagnosis of "kidney injury". Within these two groups the polypeptides are first classified according to whether it is a "positive" one, that is, it occurs multiplied in case of illness, or of a "negative" one. The polypeptides are then classified according to their mass in an ascending sequence. In the following table the individual columns have the following meaning: Table (continued 1) Table (continued 2) Table (continuation 3) Table (continued 4) Table (continued 5) Table (continued 6) Table (continued 7) Table (continued 8) Table (continued 9) Table (continued 10) Table (continued 11) Table (continued 12) Table (continued 13) Table (continued 14) Table (continuation 15) Table (continued 16) Table (continued 17) In a preferred embodiment of the invention, the device according to the invention or the method according to the invention is further characterized in that the polypeptides are defined by the indication of their relevant mass and their retention time in relevant capillary electrophoresis., as can be determined with capillary electrophoresis coupled to a mass spectrometer. In a further preferred embodiment the data bank for diagnosis "diabetes" comprises at least one, a sub-combination or all data records of polypeptides No. 1 to No. 157 of the preceding table. In another preferred embodiment, the data bank for the diagnosis "kidney injury" comprises at least one, a sub-combination or all the data records of the polypeptides No. 158 to 380 of the preceding table. In a preferred embodiment of the invention, the marker according to the invention is further characterized in that the polypeptides are defined by the indication of their relevant mass and their retention time in relevant capillary electrophoresis, as determined by coupled capillary electrophoresis to a mass spectrometer. In particular, the marker is characterized in that for the diagnosis "diabetes" there is (are) at least one polypeptide, a sub-combination of the polypeptides or all the polypeptides of No. 157 of the preceding table, or for diagnosis " kidney injury "at least one polypeptide, a subcombination of the polypeptides or all polypeptides No. 158 to 380 of the preceding table.

In a particularly preferred embodiment of the invention it is possible to use combinations of particularly preferred polypeptides for the device, method or marker. As "positive" diabetes polypeptides, polypeptides No. 32 (A), 1 (B), 48 (C), 2 (D), 44 (E), 22 (F), 9 (G), 23 ( H) and 20 (1) and their combinations, in particular as indicated below. As "negative" diabetes polypeptides, polypeptides No. 123 (A), 153 (B), 155 (C), 105 (D), 150 (E), 121 (F), 157 (G), 92 ( H) and 69 (1) and their combinations, in particular as indicated below. As polypeptides of "positive" nephropathy, polypeptides No. 225 (A), 208 (B), 164 (C), 166 (D), 171 (E), 204 (F), 206 (G), 182 ( H) and 210 (1) and their combinations, in particular as indicated below. As "negative" nephropathy polypeptides, polypeptides No. 262 (A), 260 (B), 306 (C), 358 (D), 279 (E), 318 (F), 305 (G), 261 (H) are preferred. ) and 278 (I) and combinations thereof, in particular as indicated below In another still more preferred embodiment, two of the preferred polypeptides A, B, C, D, E, F, G, H and I mentioned above as polypeptides of "negative" diabetes, "positive" diabetes, "negative" nephropathy or "positive" nephropathy. In particular, these are the combinations of the polypeptides: - and B, A and C, A and D, A and E, A and F, A and G, A and H, A and I, - B and C, B and D, B and E, B and F, B and G, B and H, B el, - C and D, C and E, C and F, C and G, C and H, C and I, - D and E, D and F, D and G, D and H, D and I, - E and F, E and G, E and H, E and I, - F and G, F and H, F, G and H, G, or H and I. In another further preferred embodiment of the invention, three of the preferred polypeptides A, B, C, D, E, F, G, H are used. I previously cited as polypeptides of "negative" diabetes, "positive" diabetes, "negative" nephropathy or "positive" nephropathy. In particular, these are the combinations of the polypeptides: A and B in combination with one of the polypeptides C, D, E, F, G, H or I, A and C in combination with one of the polypeptides D, E, F, GH or I, - A and D in combination with one of the polypeptides E, F, GH or I, - A and E in combination with one of the polypeptides F, G, H or I, - A and F in combination with one of the polypeptides G, H or I, - A and G in combination with one of the polypeptides H or I, - A and H in combination with the polypeptide I, - B and C in combination with one of the polypeptides D, E, F, G, H or I, - B and D in combination with one of the polypeptides E, F, G , H or I, - B and E in combination with one of the polypeptides F, G, H or I, - B and F in combination with one of the polypeptides G, H or I, - B and G in combination with one of the polypeptides H or I, - B and H in combination with I, - C and D in combination with one of the polypeptides E, F, G, H or I, - C and E in combination with one of the polypeptides F, G, H or I, - C and F in combination with one of the G, H or I polypeptides, - C and G in combination with one of the polypeptides H or I, - C and H in combination with I, - D and E in combination with one of the F, G, H or I, - D and F polypeptides in combination with one of the G, H or I polypeptides, - D and G in combination with one of the polypeptides H or I, - D and H in combination with I, - E and F in combination with one of the polypeptides G, H or I, - E and G in combination with one of the polypeptides H or I, - E and H in combination with I, - F and G in combination with H or I, - F and H in combination with I or - G and H in combination with I. In a further preferred embodiment of the invention, four of the preferred polypeptides A, B, C, D, E, F, G, H and I mentioned above are used as polypeptides of "negative" diabetes, of "positive" diabetes, of "negative" nephropathy or "positive" nephropathy. In particular, these are the combinations of the polypeptides: - A, B and C in combination with one of the polypeptides D, E, F, G, H or I, - A, B and D in combination with one of the polypeptides E, F, G, H or I, - A, B and E in combination with one of the polypeptides F, G, H or I, - A, B and F in combination with one of the polypeptides G, H or I, - A, B and G in combination with one of the polypeptides H or I, - A, B and H in combination with I, - A, C and D in combination with one of the polypeptides E, F, G, H or I, - A, C and E in combination with one of the polypeptides F, G, H or I, - A, C and F in combination with one of the polypeptides G, H or I, - A, C and G in combination with one of the polypeptides H or I, - A, C and H in combination with I, - A, D and E in combination with one of the polypeptides F, G, H or I, - A, D and F in combination with one of the polypeptides G, H or I, - A, D and G in combination with one of the polypeptides H or I, - A, D and H in combination with I, - A, E and F in combination with one of the polypeptides G, H or I, - A, E and G in combination with one of the polypeptides H or I, - A, E and H in combination with I, - A, F and G in combination with one of the polypeptides H or I, - A, F and H in combination with I, - A, G and H in combination with I, - B, C and D in combination with one of the polypeptides E, F, G, H or I, - B, C and E in combination with one of the polypeptides F, G, H or I, - B, C and F in combination with one of the polypeptides G, H or I, - B, C and G in combination with one of the polypeptides H or I, - B, C with H in combination with I, - B, D and E in combination with one of the polypeptides F, G, H or I , - B, D and F in combination with one of the polypeptides G, H or - B, D and G in combination with one of the polypeptides H or I, - B, D and H in combination with I, - B, E and F in combination with one of the polypeptides G, H or I, - B, E and G in combination with one of the polypeptides H or I, - B, E and H in combination with I, - B, F and G in combination with one of the polypeptides H or I, - B, F and H in combination with I, - B, G and H in combination with I, - C, D and E in combination with one of the polypeptides F, G, H or I, - C, D and F in combination with one of the polypeptides G, H or I, - C, D and G in combination with one of the polypeptides H or I, - C, D and H in combination with I, - C, E and F in combination with one of the polypeptides G, H or I, - C, E and G in combination with one of the polypeptides H or I, - C, E and H in combination with I, - C, F and G in combination with one of the polypeptides H or I, - C, F and H in combination with I, - C, G and H in combination with I, - D, E and F in combination with one of the polypeptides G, H or I, - D, E and G in combination with one of the polypeptides H or I, - D, E and H in combination with I, - D, F and G in combination with one of the polypeptides H or I, - D, F and H in combination with I, - D, G and H in combination with I, - E, F and G in combination with one of the polypeptides H or I, - E, F and H in combination with I, - E, G and H in combination with I or - F, G and H in combination with I. In another further preferred embodiment of the invention five of the preferred polypeptides A, B, C, D, E, F are used , G, H and I mentioned above as polypeptides of "negative" diabetes, "positive" diabetes, "negative" nephropathy or "positive" nephropathy. In particular, these are the combinations of the polypeptides: - A, B, C and D in combination with one of the polypeptides? F, G, H or I, - A, B, C and E in combination with one of the F polypeptides, G, H or I, - A, B, C and F in combination with one of the polypeptides G, H or I, -A, B, CyGen in combination with one of the polypeptides H or I, A, B, C and H in combination with the polypeptide I, A, B, D and E in combination with one of the polypeptides F, G, Go I, A, B D and F in combination with one of the G, H or I polypeptides, A, B, D and G in combination with one of the polypeptides H or I, A, B, D and H in combination with the polypeptide I, A, B, E and F in combination with one of the polypeptides G, H or I, A, B, E and G in combination with one of the polypeptides H or I, A, B, E and H in combination with the polypeptide I, A, B, F and G in combination with one of the polypeptides H or I, A, B, F and H in combination with polypeptide I, A, B, G and H in combination with polypeptide I, A, c, D and E in combination with one of the polypeptides F, G, H or I, A, c, D and F in combination with one of the G, H or I polypeptides, A, c, D and G in combination with one of the polypeptides H or I, A, c, D and H in combination with polypeptide I, A, c, E and F in combination with one of the polypeptides G, Gol, A, c, E and G in combination with one of the polypeptides H or I, A, c, E and H in combination with polypeptide I, A, c, F and G in combination with one of the polypeptides H or I, A, c, F and H in combination with polypeptide I, A, c, G and H in combination with polypeptide I, A, D, E and F in combination with one of the G, H or I polypeptides, A, D, E and G in combination with one of the polypeptides H or I, A, D, E and H in combination with polypeptide I, A, D, F and G in combination with one of the polypeptides H or I, - A, D, F and H in combination with polypeptide I, - A, D, G and H in combination with polypeptide I, - A, E, F and G in combination with one of the polypeptides H or I, - A, E, F and H in combination with polypeptide I, - A, E, G and H in combination with polypeptide I, - A, F, G and H in combination with polypeptide I, - B, C, D and E in combination with one of the F, G, H or I polypeptides, - B, C, D and F in combination with one of the G, H or I polypeptides, - B, C, D and G in combination with one of the polypeptides H or I, - B, C, D and H in combination with polypeptide I, - B, C, E and F in combination with one of the G polypeptides , H or I, - B, C, E and G in combination with one of the polypeptides H or I, - B, C, E and H in combination with the polypeptide I, - B, C, F and G in combination with one of the H polypeptides or I, - B, C, F and H in combination with the polypeptide I, - B, C, G and H in combination with the polypeptide I, - B, D, E and F in combination with one of the G polypeptides, H or I, - B, D, E and G in combination with one of the polypeptides H or I, - B, D, E and H in combination with polypeptide I, - B, D, F and G in combination with one of the H polypeptides or I, - B, D, F and H in combination with polypeptide I, - B, D, G, and H in combination with polypeptide I, - B, E, F and G in combination with one of the H polypeptides or I, - B, E, F and H in combination with polypeptide I, - B, E, G and H in combination with polypeptide I, - B, F, G and H in combination with polypeptide I, - C, D, E and F in combination with one of the G, H or I, C, D, E and G polypeptides in combination with one of the H or polypeptides.

I, - C, D, E and H in combination with the polypeptide I, - C, D, F and G combination with one of the polypeptides H or I, - C, D, F and H in combination with the polypeptide I, - C, D, G and H in combination with the polypeptide I, - C, E, F and G combination with one of the polypeptides H or I, - C, E, F in combination with polypeptide I, - C, E, G and H in combination with polypeptide I, - C, F, G and H in combination with polypeptide I, - D, E, F and G in combination with one of the polypeptides H or I, -D, E, F and H in combination with polypeptide I, -D, E, G and H in combination with polypeptide I, -D, F, G and H in combination with polypeptide I or-E, F, G and H in combination with polypeptide I. In another further preferred embodiment of the invention six of the preferred polypeptides A, B, C, D, E, F, G, H and I mentioned above as polypeptides are used. of "negative" diabetes, "positive" diabetes, "negative" nephropathy, or "positive" nephropathy. In particular, these are the combinations of the polypeptides: - A, B, C, D and E in combination with one of the polypeptides F, G, H or I, - A, B, C, D and F in combination with one of the polypeptides G, H or I, - A, B, C or G in combination with one of the polypeptides H or I, - A, B, C, D and H in combination with polypeptide I, - A, B, C, E and F in combination with one of the polypeptides G, H or I, A, B, C, E and G in combination with one of the polypeptides H or I, - A, B, C, E and H in combination with the polypeptide I, - A, B, C, F in combination with one of the polypeptides H or I, - A, B, C, F in combination with polypeptide I, - A, B, C, G and H in combination with polypeptide I, - A, B, D, E in combination with one of the G polypeptides, H or I, - A, B, D, E and G in combination with one of the polypeptides H or I, - A, B, D,? and H in combination with the polypeptide I, - A, B, D, F and G in combination with one of the polypeptides H or I, - A, B, D, F and H in combination with polypeptide I, - A, B, D, G and H in combination with the polypeptide I, - A, B, E, F and G in combination with one of the polypeptides H or I, - A, B, E, F and H in combination with the polypeptide I, - A, B, E, G and H in combination with polypeptide I, - A, B, F, G and H in combination with polypeptide I, - A, C, D, E and F in combination with one of polypeptides G, H or I, - A, C, D, E and G in combination with one of the polypeptides H or I, - A, C, D, E and H in combination with polypeptide I, - A, C, D , F and G in combination with one of the polypeptides H or I, - A, C, D, F and H in combination with polypeptide I, - A, C, D, G and H in combination with polypeptide I, - A, C, E, F and G in combination with one of the polypeptides H or I, - A, C, E, F and H in combination with polypeptide I, - A, C, E, G and H in combination with the polypeptide I, - A, C, F, G and H in combination with the polypeptide I, - A, D, E, F and G in combination with one of the polypeptides H or I, - A, D, E, F and H in combination with polypeptide I, _ A, D, E, G and H in combination with polypeptide I, - A, D, F, G and H in combination with polypeptide I, - A, E, F, G and H in combination with polypeptide I, - B, C, D, E and F in combination with one of the G, H or I polypeptides, - B, C, D, E and G in combination with one of the polypeptides H or I, - B, C, D, E and H in combination with polypeptide I, - B, C, D, F and G in combination with one of the polypeptides H or I, - B, C, D, F and H in combination with polypeptide I, - B, C, D, G and H in combination with polypeptide I, - B, C, E, F and G in combination with one of the H polypeptides or I, - B, C, E, F in combination with polypeptide I, - B, C, E, GyH in combination with polypeptide I, - B, C, FG and H in combination with polypeptide I, - B , D, E, F and G in combination with one of the polypeptides H or I, - B, D, E, F and H in combination with polypeptide I, - B, D, E, GyH in combination with polypeptide I, - B, D, F, G and H in combination with polypeptide I, - B , E, F, G and H in combination with the polypeptide I, - C, D, E, F and G in combination with one of the polypeptides H or I, - C, D, E, F and H in combination with polypeptide I, - C, D, E, G and H in combination with polypeptide I, - C, D, F, G and H in combination with polypeptide I , - C, E, F, G and H in combination with the polypeptide I or - D, E, F, G and H in combination with the polypeptide I, In another further preferred embodiment of the invention seven of the Preferred polypeptides A, B, C, D, E, F, G, H and I mentioned above as polypeptides of "negative" diabetes, "positive" diabetes, "negative" nephropathy or "positive" nephropathy. In particular, these are the combinations of the polypeptides: A, B, C, D, E and F in combination with one of the polypeptides G, H or I, - A, B, C, D, E and G in combination with one of polypeptides H or I, - A, B, C, D, E and H in combination with polypeptide I, A, B, C, D, F and G in combination with one of the polypeptides H or I, - A, B, C, D, F and H in combination with polypeptide I, - A, B, C, D, G and H in combination with the polypeptide I, - A, B, C, E, F and G in combination with one of the polypeptides H or I, - A, B, C, E, F and H in combination with the polypeptide I, - A, B, C, E, G and H in combination with polypeptide I, - A, B, C, F, G and H in combination with polypeptide I, - A, B, D, E, F and G in combination with one of the polypeptides H or I, - A, B, D, E, F and H in combination with polypeptide I, - A, B, D, E, G and H in combination with polypeptide I , - A, B, D, F, G and H in combination with polypeptide I, - A, B, E, F, G and H in combination with polypeptide I, - A, C, D, E, F and G in combination with one of the polypeptides H or I, - A, C, D, E, F in combination with polypeptide I, - A, C, D, E, G and H in combination with polypeptide I, - A , C, D, F, G and H in combination with polypeptide I, - A, C, E, F, G and H in combination with polypeptide I, - A, D, E, F, G and H in combination with polypeptide I, - B, C, D,?, F in combination with one of the polypeptides H or I, - B, C, D, E, F and H in combination with polypeptide I, - B, C, D, E, G and H in combination with polypeptide I, - B, C, D, F, G and H in combination with polypeptide I, - B, C, E, F, G and H in combination with polypeptide I, - B, D, E, F, G and H in combination with polypeptide I or C, D, E, F, G and H in combination with the polypeptide I. In another further preferred embodiment of the invention eight of the preferred polypeptides A, B, C, D, E, F, G, H and E are used. I previously cited as polypeptides of "negative" diabetes, "positive" diabetes, "negative" nephropathy or "positive" nephropathy. In particular, these are the combinations of the polypeptides: - A, B, C, D, E, F, G and H - A, B, C, D, E, F, G and I, - A, B, C, D, E, F, H and i, - A, B, C, D, E, G, H and I, - A, B, C, D, F, G, H and I, - A, B, C, E, F, G, H and I, - A, B, D, E, F, G, H and I, - A, C, D, E, F, G, H and I or - B, C, D , E, F, G, H and I. In another further preferred embodiment of the invention, all nine of the preferred polypeptides A, B, C, D, E, F, G, H and I mentioned above are used as polypeptides of "negative" diabetes, "positive" diabetes, "negative" nephropathy or "positive" nephropathy. In particular, these are the combinations of the polypeptides: - A, B, C, D, E, F, G, H and I. It is noted that in relation to this date, the best method known by the applicant to carry the practice said invention is that which is clear from the present description of the invention.

Claims (12)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. Device for the quantitative evaluation of polypeptides contained in a body fluid sample and comparison with reference values stored in a database, characterized in that the reference values are stored as polypeptide data records relevant to the state that in each case comprise the less an indication on the probability of the occurrence or concentration of the polypeptides for a pathological state in samples from healthy and sick subjects.

Device according to claim 1, characterized in that the polypeptides are defined by the indication of their relevant mass and their relevant retention time in capillary electrophoresis, as determined by capillary electrophoresis coupled to a mass spectrometer.

Device according to claim 2, characterized in that for the diagnosis "diabetes" the data bank comprises at least one, a sub-combination or all the data records of the polypeptides according to the following table: Continuation 1 of the table Continuation 2 of the table Continuation 3 of the table Continuation 4 of the table Continuation 5 of the table Continuation 6 of the table Continuation 7 of the table

4. Device according to claim 2, characterized in that for the diagnosis "renal injury" the data bank comprises at least one, a subcombination or all data records of the polypeptides according to the following table: Continuation 1 of the table Continuation 2 of the table Continuation 3 of the table Continuation 4 of the table Continuation 5 of the table Continuation 6 of the table Continuation 7 of the table Continuation 8 of the table Continuation 9 of the table Continuation 10 of the table

5. Method for the quantitative evaluation of polypeptides contained in a sample of body fluid and comparison with reference values stored in a database, characterized in that reference values are used for comparison as records of polypeptide data relevant to the state when comparing an indication of the probability of the occurrence and / or concentration of the polypeptides in the body fluid sample with the indication in each case of at least one reference value on the probability of the occurrence and / or concentration of the polypeptides for a state pathological in samples of healthy and sick subjects.

Method according to claim 5, characterized in that the polypeptides are defined by the indication of their relevant mass and their relevant retention time in capillary electrophoresis, as determined by capillary electrophoresis coupled to a mass spectrometer.

7. Method according to claim 6, characterized in that for the diagnosis "diabetes" is used at least one, a sub-combination or all data records of the polypeptides according to the following table: Continuation 1 of the table Continuation 2 of the table Continuation 3 of the table Continuation 4 of the table Continuation 5 of the table Continuation 6 of the table Continuation 7 of the table

8. Method according to claim 6, characterized in that for the diagnosis "renal lesion" at least one, a subcombination or all data records of the polypeptides are used according to the following table: Continuation 1 of the table Continuation 2 of the table Continuation 3 of the table Continuation 4 of the table Continuation 5 of the table Continuation 6 of the table Continuation 7 of the table Continuation 8 of the table Continuation 9 of the table Continuation 10 of the table Continuation 11 of the table

9. Marker to detect disease states, characterized in that it comprises a multitude of polypeptides relevant to the state, which in each case are linked with an indication on the probability of the occurrence and / or concentration of the polypeptide for a pathological state in samples from healthy and sick subjects.

10. Marker in accordance with the claim 9, characterized in that the polypeptides are defined by the indication of their relevant mass and their retention time in relevant capillary electrophoresis, as determined with capillary electrophoresis coupled to a mass spectrometer.

11. Marker in accordance with the claim 10, characterized in that for the diagnosis "diabetes" contains at least one polypeptide, a sub-combination of the polypeptides or all the polypeptides of the following table: Continuation 1 of the table Continuation 2 of the table Continuation 3 of the table Continuation 4 of the table Continuation 5 of the table Continuation 6 of the table Continuation 7 of the table Continuation 8 of the table

12. Marker according to claim 10, characterized in that for the diagnosis "renal lesion" contains at least one polypeptide, a sub-combination of the polypeptides or all the polypeptides of the following table: Continuation 1 of the table Continuation 2 of the table Continuation 3 of the table Continuation 4 of the table Continuation 5 of the table Continuation 6 of the table Continuation 7 of the table Continuation 8 of the table Continuation 9 of the table Continuation 10 of the table