Method and system for measuring a sample to determine the presence of and optionally treat a pathologic condition

1. A method for determining the presence of at least one hormone form of an endogenous hormone that is involved in diabetes or metabolic syndrome in a sample comprising:

(a) forming within a free flow electrophoretic (FFE) chamber a separation zone between an anode and a cathode that comprises a zone I formed by at least one separation buffer medium type I (SBM type I) and a zone II formed by at least one separation buffer medium type II (SBM type II),

wherein the SBM type I yields denaturing conditions and comprises at least one additive at an effective concentration and the SBM type II yields non-denaturing conditions and does not contain the additive at the effective concentration;

(b) introducing the sample into the separation zone; and

(c) transferring the at least one hormone form from the SBM type I into the SBM type II, or transferring the at least one hormone form from the SBM type II into the SBM type I during the FFE separation, thereby separating the hormone form from an interfering form that would interfere with measuring a parameter of the hormone form by an analytical method employed in step (d),

wherein the endogenous hormone is selected from the group consisting of insulin, glucagon-like-peptide 1 (GLP-1), ghrelin, cholecystokinin (CKK), leptin, and obestatin; and

(d) measuring the parameter of the hormone form by the analytical method.

2. The method according to claim 1 , wherein the electrophoretic separation of step (c) is carried out in an operation method selected from the group consisting of zone electrophoresis, isoelectric focusing, combinations thereof, and isotachophoresis.

3. The method according to claim 1 , wherein the sample comprises a second hormone form, and

wherein the at least one hormone form and the second hormone form are quantitatively separated from each other.

4. The method according to claim 1 , wherein the hormone form is involved in diabetes type I or diabetes type II.

5. The method according to claim 1 , wherein the endogenous hormone is insulin, and wherein the at least one hormone form is selected from the group consisting of heterologous insulin, Glargine, Lispro, Aspart, Glulisine, and Detemir.

6. The method according to claim 1 wherein the analytical method in step (b) step (d) is selected from the group consisting of immuno assay, mass spectrometry, SPR, critical angle refractometry, total internal fluorescence (TIRF), total internal reflection phosphorescence, total internal reflection light scattering, evanescent wave elipsometry, Brewster angle reflectometry, circular dichroism, UV or UV-VIS-spectroscopy, IR-spectroscopy, pulsed amperometric detection, and Raman spectroscopy.

7. The method according to claim 1 wherein the analytical method in step (d) comprises online measuring of the parameter measured in step (d) and is selected from the group consisting of SPR, critical angle refractometry, total internal fluorescence (TIRF), total internal reflection phosphorescence, total internal reflection light scattering, evanescent wave elipsometry, Brewster angle reflectometry, circular dichroism, UV or UV-VIS-spectroscopy, IR-spectroscopy, pulsed amperometric detection, and Raman spectroscopy.

8. The method according to claim 1 , wherein the parameter correlates with the presence/absence, amount, concentration, weight ratio, or concentration ratio of the at least one hormone form.

9. The method according to claim 1 , wherein the analytical method is carried out to determine the concentration/amount of the at least one hormone form that is present in the sample, and

wherein the concentration/amount of the at least one hormone form that is present in the sample is determined, wherein the method further comprises:

determining the absence/presence of a second hormone form of the endogenous compound comprising simultaneously measuring by the analytical method the second hormone form, wherein the second hormone form is selected from the group consisting of another hormone involved in diabetes or metabolic syndrome, a hormone that is not involved in diabetes and metabolic syndrome, a protein, a saccharide, a nucleic acid, and a metabolite of the glucose pathway.

10. The method according to claim 1 , wherein the analytical method is a multiplexing immuno array or SPR comprising a multiplexing biosensor.

11. The method according to claim 1 , wherein the at least one hormone form is separated from an interaction partner of the at least one hormone form,

wherein the introduction step comprises introducing the sample comprising the hormone form and the interaction partner into the zone I,

wherein the presence of the at least one additive at the effective concentration in the zone I suppresses an interaction between the hormone form and the interaction partner,

wherein the interaction partner remains in the zone I, whereas the hormone form is transferred into the zone II,

wherein the zone I comprises a pH plateau,

wherein the interaction partner is albumin, and

wherein the endogenous hormone is a peptide hormone.

12. The method according to claim 3 , wherein the at least one hormone form has an amino acid sequence that differs from an amino acid sequence of another analog of the endogenous hormone by at most 5 amino acids,

wherein the at least one hormone form differs from the other analog by a modification of at least one amino acid, and

wherein the modification comprises a substituent covalently attached to the at least one amino acid that is selected from the group consisting of a C 1 -C 20 fatty acid, a C 1 -C 20 alkyl, a C 1 -C 20 alkanoyl, a monosaccharide, a disasaccharide, a trisaccharide, a polysaccharide, a phosphate, a sulfonate, and a sialyl residue.