Systems and methods using paramagnetic agents for in vitro diagnostic applications

Abstract

The present invention discloses a method and systems for detecting a target biochemical molecular species whose main component is water. In one aspect, the method comprises steps of: (a) obtaining a sample whose main component is water; (b) providing Functionalized Paramagnetic Particles (FPP) comprising a paramagnetic core and a moiety configured to interact with the target biochemical molecular species; (c) contacting the FPP with the sample; (d) exposing the sample to an applied magnetic field; (e) measuring a change in a nuclear relaxation property of the sample, caused by the interaction between the FPP and the biochemical molecular species in the applied magnetic field; and (f) correlating the change to the presence of the biochemical molecular species in the sample. According to a main aspect of the invention, a change in T.sub.1 nuclear relaxation property of the water protons in the sample is correlated to the presence of the target biochemical molecular species.

What is claimed is: 
     
         1 . A method for detecting a target biochemical molecular species in a sample whose main component is water, comprising the steps of:
 a. obtaining a sample whose main component is water;   b. providing Functionalized Paramagnetic Particles (FPP) comprising a paramagnetic core and a moiety configured to interact with said target biochemical molecular species;   c. contacting said FPP with said sample;   d. exposing said sample to an applied magnetic field;   e. measuring a change in a nuclear relaxation property of said sample, caused by said interaction between said FPP and said biochemical molecular species in the applied magnetic field with a magnetic resonance device (MRD); and   f. correlating said change to the presence of said biochemical molecular species in said sample;   
       wherein a change in T.sub.1 nuclear relaxation property of the water protons in said sample is correlated to the presence of said target biochemical molecular species; and 
       wherein said FPP comprises a non ferrous oxide paramagnetic core. 
     
     
         2 . The method according to  claim 1 , comprising at least one additional step selected from the group consisting of:
 performing two or more measurements to determine the relaxation time of the sample, wherein the measurements are performed before and after at least one addition of said FPP;   detecting said target biochemical molecular species in vitro;   forming said FPP as a single molecule, a multimeric system, a micro-sized vesicle or particle, a nano-sized vesicle or particle, a liposome, a probe or any combination thereof;   selecting said sample from a group consisting of a liquid, a gas, a slurry, a liquid containing particulates, a gas containing particulates, a gel, a sol, a suspension, a solution, a dispersion, a colloid, a mixture, an emulsion, an aerosol, a liquid containing solid objects, a gas containing solid objects, and any combination thereof;   selecting said sample from a group comprising a biological fluid, a biological tissue, a tissue extract, an industrial fluid, food sample, a beverage, wine, water, potable water, sewage, irrigation water, sea water, river water, lake water, industrial effluent, farm effluent, effluent from human habitation, road runoff, cleaning fluid, a gas sample or any combination thereof;   selecting said biological fluid from a group comprising urine, blood, lymph, plasma, cerebrospinal fluid, saliva, amniotic fluid, bile and tears;   providing said sample within a production process in an industry selected from pharmaceuticals production, food production, beverage production, chemical refining, chemical processing, medical products, biological products, metal casting, metal refining, desalination, fluid purification, and sewage processing;   applying a magnetic field, thereby enhancing the change in a paramagnetic nuclear relaxation property of said sample upon comparing relaxation rates at two different magnetic fields;   measuring a change in a nuclear relaxation property of said sample using a portable NMR or MRI measuring means;   measuring a change in a nuclear relaxation property of said sample using a magnetic resonance device (MRD) consisting of magnets housed within a cage; and   measuring a change in a nuclear relaxation property of said sample using a self-fastening cage type of a magnetic resonance device (MRD).   
     
     
         3 . A method according to  claim 1 , wherein said change in nuclear relaxation property is measured using a self-fastening cage magnetic resonance device (MRD), and comprising at least one additional step selected from the group consisting of:
 providing a homogeneous, stable and uniform magnetic field therein, further wherein said self-fastening cage type MRD is additionally characterized by an outside shell comprising at least three flexi-jointed superimposed walls;   providing an MRD characterized by an outside shell, said outside shell comprising at least three flexi-jointed superimposed walls disposed in a predetermined clockwise or counterclockwise arrangement, said MRD comprising at least six side-magnets arranged in two equal groups being in a face-to-face orientation in a magnetic connection with said outside shell, increasing the overall strength of the magnetic field provided in said cage; at least two pole-magnet pieces, arranged in a face-to-face orientation in between said side-magnets; at least two main-magnets, located on said pole-pieces, arranged in a face-to-face orientation, generating the static magnetic field in said cage; and shimming mechanism, said mechanism selected from the group consisting of an array of active shim coils, passive shimming elements or a combination thereof; wherein at least a portion of said side-magnets ( 2 ) are superconductors or ferromagnets;   providing a magnetic resonance device adapted to producing high contrast high resolution images of said sample;   providing a magnetic resonance device comprising: an envelope for least partially confining said sample; a plurality of magnets located at least partially around said envelope, said plurality of magnets comprising: a least one first magnet configured to provide a high magnetic field for generating multiple time-resolved one or more first images at high resolution of at least a portion of said sample; and a least one second magnet configured to provide a low magnetic field for generating multiple time-resolved one or more second images at high contrast of at least portion of same said sample; wherein at least one image of said first images and at least one image of said second images being generated in a time no greater than approximately the time between two first images; and, a CPU to process said images comprising a computer readable medium containing instructions for generating at least one third image superimposing at least one image of said first images with at least one image of said second images;   generating multiple time resolved one or more first images at high resolution of at least a portion of said sample; generating multiple time resolved one or more second images at high contrast of at least portion of same said sample; and then superimposing at least one image of said first images with at least one image of said second images; whereby a high-contrast, high resolution real-time continuous image of said sample is obtained;   selecting said at least one first magnet to be of 2 Tesla and lower;   selecting said at least one first magnet to be of 2 Tesla and higher; said at least one first magnet from a group consisting of permanent magnets, electromagnets, superconducting magnets, and any combination thereof;   selecting said at least one second magnet to be of 2 Tesla and lower;   selecting said at least one second magnet to be of 2 Tesla and higher;   selecting said at least one second magnet from a group consisting of permanent magnets, electromagnets, superconducting magnets, and any combination thereof;   generating a magnetic resonance signal in the range of about 0.1 Tesla and about 10 Tesla;   generating a magnetic resonance signal in the range of 2 Tesla and lower;   generating a magnetic resonance signal in the range of 2 Tesla and higher;   applying a magnetic resonance frequency in the range of about 5 MHz to about 40 MHz;   selecting said magnets from a group consisting of permanent magnets, electromagnets, superconducting magnets, and any combination thereof;   selecting said moiety from a group comprising antibodies, antibody fragments, monoclonal antibody, receptors, ligands, macromolecules, peptides, hormones, fatty acids, lipids, receptor agonists and antagonists, amino acids, sugars, lectins, albumins, polycarbon molecules, glycoproteins, nucleic acids, pegylated molecules, liposomes, chelators, cells, viruses, chemotherapeutic agents and any combination thereof;   selecting said target biochemical molecular species from a group comprising a biological molecule, a chemical molecule, an analyte, a contaminant, a particle, a pathogen or any combination thereof;   selecting said target biochemical molecular species from a group comprising a protein, a pathogen, a prion, a virus, a bacteria, a contaminant, a pathological isoform, a biomarker, an allergen, a neurotransmitter, an antigenic determinant, an epitope, a cell marker, cell membrane marker or epitope, a membrane marker, an enzyme, a chemical molecule, an analyte, a receptor, a ligand, a macromolecule, a peptide, a hormone, a fatty acid, a lipid, a receptor agonist and antagonist, an amino acid, a sugar, a glycoprotein, a nucleic acid, an antioxidant agent, a chemotherapeutic agent, a biological tissue and any combination thereof;   selecting said analyte from a group comprising an organic analyte and an inorganic analyte;   selecting said inorganic analyte from a group comprising molecular oxygen, oxygen-containing radicals and combinations thereof; detecting oxygen-containing radicals “ad hoc” generated, for assessing the anti-oxidant properties of the sample;   selecting said paramagnetic core as a metal ion, a metal complex, oxides of a metal ion, oxides of a transition metal, mixed oxides of a transition metal and their mixtures;   selecting said paramagnetic core from a group comprising metal complexes, aggregates of metal complexes, polymer-bound metal complexes, stable organic radicals and any combination thereof;   selecting said metal ion from a group comprising an ion of nickel, iron, manganese, copper, gadolinium, europium and mixtures thereof;   assessing redox characteristics, comprising steps of detecting differences in Paramagnetic Relaxation Enhancement (PRE) properties induced by a change in at least one redox characteristic of said FPP, using an applied magnetic field;   selecting said redox characteristic from a group comprising lipid peroxidation, lipid peroxidation followed by a change in membrane permeability, redox potential of metal ions, formation and cleavage of disulfide bonds, oxidation state, antioxidant activity and any combination thereof;   providing said FPP as a liposome loaded with a plurality of paramagnetic payloads;   applying a magnetic field, thereby enhancing a change in the permeability of said liposome so as to affect a nuclear relaxation property of said sample;   applying a magnetic field, thereby enhancing a change in at least one of cleavage or formation of disulfide bonds of said moiety so as to affect a nuclear relaxation property of said sample;   applying a magnetic field, thereby affecting at least one property of said FPP selected from a group comprising concentration, lipid peroxidation, membrane permeability, redox potential, formation and cleavage of disulfide bonds, oxidation state, redox potential, activation state, binding affinity, and any combination thereof, so as to induce a change in a nuclear relaxation property of said sample;   conjugating said liposome with a site-specific ligand;   conjugating said liposome with a biotin activated molecule; and   providing said FPP as a biotinylated liposome.   
     
     
         4 . A method for detection of a biomarker in a sample whose main component is water, comprising the steps of:
 obtaining a sample whose main component is water;   providing liposomes loaded with a plurality of paramagnetic agents, said liposomes conjugated with a site specific moiety configured to interact with said biomarker in said sample;   contacting said liposomes with said sample under conditions that allow the interaction between the site specific moiety and said biomarker;   exposing said sample to an applied magnetic field; and   measuring a change in a nuclear relaxation property of said sample caused by said interaction between the liposomes and said biomarker in the applied magnetic field;   wherein a change in T.sub.1 nuclear relaxation property is correlated to the presence of said biomarker in said sample.   
     
     
         5 . The method according to  claim 4 , comprising additional steps of:
 providing biotinylated liposomes; said liposomes loaded with a plurality of paramagnetic agents;   providing biotinylated ligands, said ligands configured to interact with said biomarker;   providing activated avidin molecules;   contacting said biotinylated liposomes, said biotinylated ligands and said activated avidin molecules with said sample so as to enable avidin-biotin interaction, thereby forming complexes comprising said liposomes, said ligand and said biomarker; such that said complexes are specific to said biomarker; and,   measuring a change in a nuclear relaxation property of said sample caused by said complex formation in the applied magnetic field;   wherein a change in T.sub.1 nuclear relaxation property is correlated to the presence of said biomarker in said sample.   
     
     
         6 . The method according to  claim 4 , wherein said sample is selected from a group consisting of a liquid, a gas, a slurry, a liquid containing particulates, a gas containing particulates, a gel, a sol, a suspension, a solution, a dispersion, a colloid, a mixture, an emulsion, an aerosol, a liquid containing solid objects, a gas containing solid objects, and any combination thereof. 
     
     
         7 . A system for detection of a biomarker in a sample whose main component is water, comprising:
 a. a sample whose main component is water;   b. liposomes loaded with a plurality of paramagnetic agents, said liposomes conjugated with a site specific moiety configured to interact with said biomarker in said sample;   c. a magnetic resonance device (MRD) configured to measure a change in a nuclear relaxation property of the sample whose main component is water removed from a production batch or continuous flow of said FBW or biological fluid;   
       wherein a change in T.sub.1 nuclear relaxation property of the water protons in said sample is correlated to the presence of said biomarker in said sample. 
     
     
         8 . A method of establishing the redox properties of a production batch or continuous flow of a Foodstuff, Beverage or Wine (FBW) or of a biological fluid, comprising the steps of:
 a. obtaining a sample removed from said production batch or continuous flow of said FBW or from said biological fluid;   b. providing Functionalized Paramagnetic Particles (FPP) configured to change their redox property upon interaction with dissolved molecular oxygen or ad hoc generated radicals of said removed sample;   c. contacting said FPP with said removed sample;   d. exposing said removed sample to an applied magnetic field; and,   e. measuring a change in a nuclear relaxation property of said removed sample caused by said change in the redox properties of said FPP in the applied magnetic field;   
       wherein a change in T.sub.1 nuclear relaxation property is correlated with the presence and/or concentration of said molecular oxygen or of the ad hoc generated radicals of said removed sample, thereby establishing the redox properties of said production batch or continuous flow of said FBW or of said biological fluid. 
     
     
         9 . The method according to  claim 8 , wherein said method further comprises at least one step selected from the group consisting of the following:
 d. providing said FPP conjugated with at least one moiety configured to interact with dissolved molecular oxygen or with ad hoc generated radicals;   e. selecting said moiety selected from the group consisting of a lipid, a fatty acid, an amino acid, a peptide, a protein, a molecule containing at least one disulfide bond, a liposome, and any combination thereof;   f. measuring a change in a nuclear relaxation property of said sample caused by a change in the antioxidant activity of said removed sample in the applied magnetic field;   g. measuring a difference in Paramagnetic Relaxation Enhancement (PRE) property of said removed sample caused by said interaction of said FPP with said dissolved molecular oxygen or with said “ad hoc” generated radicals;   h. contacting said removed sample with FPP configured to form a liposome structure, said FPP comprising a paramagnetic core and a fatty acid moiety, wherein peroxidation of said fatty acid moiety substantially changes the permeability of said liposome;   i. measuring a change in a nuclear relaxation property of said removed sample based on competition for molecular oxygen consumption; providing said FPP comprising a paramagnetic core selected from a group comprising a metal ion, a metal complex, oxides of a metal ion, oxides of a transition metal, mixed oxides of a transition metal and their mixtures;   j. selecting said metal ion from a group comprising an ion of nickel, iron, manganese, copper, gadolinium, dysprosium, europium and mixtures thereof; and   k. selecting said biological fluid from a group comprising urine, blood, lymph, plasma, cerebrospinal fluid, saliva, amniotic fluid, bile and tears.   
     
     
         10 . A system for establishing the redox properties of a production batch or continuous flow of a Foodstuff, Beverage, Wine (FBW) or of a biological fluid, comprising:
 l. a magnetic resonance device (MRD) configured to measure a change in a nuclear relaxation property of a sample removed from said production batch or continuous flow of said FBW or of said biological fluid; and   m. a plurality of Functionalized Paramagnetic Particles (FPP) configured to be in contact with said removed sample, said plurality of FPP are further configured to change at least one of their redox properties upon interaction with dissolved molecular oxygen or with “ad hoc” generated radicals of said removed sample;   
       wherein a change in T.sub.1 nuclear relaxation property measured by said MRD is correlated to the presence or concentration of dissolved molecular oxygen or “ad hoc” generated radicals of said removed sample, thereby the redox properties of said production batch or continuous flow of said FBW or of said biological fluid are established. 
     
     
         11 . The system according to  claim 10 , wherein said biological fluid is selected from the group consisting of urine, blood, lymph, plasma, cerebrospinal fluid, saliva, amniotic fluid, bile and tears. 
     
     
         12 . A system for establishing the potability of a production batch or continuous flow of a flowable Foodstuff, Beverage or Wine (FBW), comprising:
 a. a magnetic resonance device (MRD) configured to measure a change in nuclear relaxation property of a sample removed from said production batch or continuous flow of said flowable FBW; and,   b. a plurality of functionalized paramagnetic particles (FPP) configured to be in contact with said sample, said plurality of FPP are further configured to change their redox/oxidative properties upon interaction with dissolved molecular oxygen or ad hoc generated radicals of said removed sample;   
       wherein a change in T.sub.1 nuclear relaxation property measured by said MRD correlates with dissolved molecular oxygen or ad hoc generated radicals concentration of said FBW sample, thereby establishing the potability of said production batch or continuous flow of said flowable FBW. 
     
     
         13 . The system according to  claim 12 , wherein at least one of the following is held true:
 a. a concentration value lower than about 6 ml of dissolved oxygen per liter of said removed sample is indicative of the potability of said production batch or continuous flow of said FBW;   b. a value lower than X ml of oxygen per liter is correlated to >7 of the 9 point hedonic scale;   c. a value lower than X ml of oxygen per liter is correlated to >y of the hybrid scale; and   d. a value lower than X ml of oxygen per liter is correlated to >y of the self-adjusting scale.