Systems and methods for characterization of molecules

Abstract

The present invention generally provides systems and methods for the detection, identification, or characterization of differences between properties or behavior of corresponding species in two or more mixtures comprised of molecules, including biomolecules and/or molecules able to interact with biomolecules, using techniques such as partitioning. The experimental conditions established as distinguishing between the mixtures of the molecules using the systems and methods of the invention can also be used, in some cases, for further fractionation and/or characterization of the biomolecules and/or other molecules, using techniques such as single-step or multiple-step extraction, and/or by liquid-liquid partition chromatography. The methods could also be used for discovering and identifying markers associated with specific diagnostics, and can be used for screening for such markers once discovered and identified during diagnostics screening.

1 . A method for identifying one or more tools for physiological analysis, comprising:
 partitioning a protein contained within a first mixture of species in at least a first phase and a second phase of a first partitioning system, wherein the second phase is immiscible with the first phase at equilibrium;   determining a relative measure of interaction between the protein and the first and second phases of the first partitioning system;   determining a relative measure of interaction between the protein contained within a second mixture of species, and the first and second phases of the first partitioning system;   determining a difference in the relative measure of interaction of the protein of the first mixture of species with the first and second phases of the first partitioning system, versus the relative measure of interaction of the protein of the second mixture of species with the first and second phases of the first partitioning system   
     
     
         2 . A method as in  claim 1 , further comprising:
 selecting, based upon the difference, the first partitioning system as a tool for determining a physiological condition of a biological system based upon determination of a relative measure of interaction between at least one species of a sample from the biological system and the first and second phases of the first partitioning system.   
     
     
         3 . A method as in  claim 1 , wherein the first partitioning system is an aqueous multi-phase system. 
     
     
         4 . A method as in  claim 1 , comprising selecting the first partitioning system as a tool for determining a physiological condition of a biological system based upon determination of a relative measure of interaction between the protein and the first partitioning system. 
     
     
         5 . A method as in  claim 1 , wherein the first mixture of species and the second mixture of species respectively comprise a sample indicative of an abnormal condition and a control sample, both from a single organism. 
     
     
         6 . A method as in  claim 5 , wherein the samples are taken from the organism at the same time. 
     
     
         7 . A method as in  claim 5 , wherein the samples are taken from the organism at a different time. 
     
     
         8 . A method as in  claim 1 , further comprising determining a difference in the relative measure of interaction of the protein contained within the first mixture of species versus the relative measure of interaction of the protein contained within the second mixture of species, in both the first partitioning system and a different, second partitioning system. 
     
     
         9 . A method as in  claim 1 , comprising determining a difference in the relative measure of interactions of the protein contained within the first mixture of species versus the relative measure of interaction of the protein contained within the second mixture of species, in only a single partitioning system. 
     
     
         10 . A method as in  claim 1 , comprising:
 determining a relative measure of interaction between the protein contained within the first mixture of species and at least first and second interacting components defining at least a first phase and a second phase, respectively, of a second system;   determining a relative measure of interaction between the protein contained within the second mixture of species and the second system;   determining a difference in the relative measure of interaction of the protein contained within the first mixture of species, versus the protein contained within the second mixture of species, with the second system; and   based upon the difference, selecting the second system as a tool, in conjunction with the first partitioning system, for determining the physiological condition.   
     
     
         11 . A method as in  claim 1 , comprising:
 determining a relative measure of interaction between the protein contained within the first mixture of species and at least first and second interacting components defining at least a first phase and a second phase, respectively, of a third system;   determining a relative measure of interaction between the protein contained within the second mixture of species and the third system;   determining a lack of significant difference in the relative measure of interaction of the protein contained within the first mixture of species, versus the protein contained within the second mixture of species, with the third system; and   based upon the lack of significant difference, rejecting the third system as a tool for determining the physiological condition.   
     
     
         12 . A method as in  claim 1 , wherein the protein contained within the first mixture of species is obtained from a biological system with a first physiological condition, and the protein contained within the second mixture of species is obtained from the same biological system with a second physiological condition. 
     
     
         13 . A method as in  claim 1 , wherein biological systems from which the first and second mixture of species are obtained represent the same individual member. 
     
     
         14 . A method as in  claim 13 , wherein the biological systems from which the first and second mixture of species are obtained represent the same individual member and are taken at different times. 
     
     
         15 . A method as in  claim 14 , comprising determining a progression of change of a relative measure of interaction using the difference in the relative measure of interaction. 
     
     
         16 . A method as in  claim 1 , wherein the biological systems from which the first and second mixtures of species are obtained represent the same species but not the same individual member. 
     
     
         17 . A method as in  claim 1 , wherein the first mixture of species is taken from a sample of a biological system having a first physiological condition, the method comprising:
 determining a relative measure of interaction between at least a second species of the first mixture of species, obtained from the biological system, with the first physiological condition and at least the first and second interacting components of the first partitioning system;   determining a relative measure of interaction between at least a second species of the second mixture of species, corresponding to the second species of the first mixture of species, obtained from the biological system, with a second physiological condition, and the first partitioning system;   determining a difference in the relative measure of interaction of the at least one second species of the first mixture of species, versus the at least one second species of the second mixture of species, with the first partitioning system; and   denoting the difference associated with the at least one second species as a tool, in conjunction with the difference in the protein, for determining a difference between the physiological conditions of the first and second biological systems.   
     
     
         18 . A method as in  claim 17 , comprising:
 determining a relative measure of interaction between at least a third species of the first mixture of species, obtained from the biological system, and with the first physiological condition, and at least the first and second interacting components of the first partitioning system;   determining a relative measure of interaction between at least the third species of the second mixture of species, obtained from the biological system, and with the second physiological condition, and the first partitioning system;   determining a lack of significant difference in the relative measure of interaction of the at least one third species of the first mixture of species, versus the at least one third species of the second mixture of species, with the first partitioning system; and   based upon the lack of significant difference, rejecting the third species as a tool for determining a difference between the physiological conditions of the first and second biological systems.   
     
     
         19 . A method as in  claim 1 , wherein the protein contained within the first mixture of species and the protein contained within the second mixture of species are chemically identical. 
     
     
         20 . A method as in  claim 1 , wherein following the selection of the first partitioning system as a tool, the relative measure of interactions are stored for later determination of an unknown sample as belonging to the physiological condition of the biological system. 
     
     
         21 . A method of determining a physiological condition of a biological system comprising:
 determining a relative measure of interaction between a protein arising from a sample from a biological system, and at least first and second interacting components defining at least a first phase and a second phase, respectively, of a first partitioning system;   from the process of determining the relative measure of interaction between the protein and the first and second interacting components of the first partitioning system, determining the physiological condition of the biological system.   
     
     
         22 . A method as in  claim 21 , wherein the method comprises:
 comparing the relative measure of interaction of the protein from the sample with a relative measure of interaction of at least one marker from a different sample from the same biological system,   based on the degree of similarity between the interaction parameters, determining a physiological condition of the biological system.   
     
     
         23 . A method as in  claim 21 , comprising comparing the relative measures of interaction of the protein and a second species with the first system with relative measures of interaction of the protein and a second species of a control with the first partitioning system.