Methods for reducing the range in concentrations of analyte species in a sample

Abstract

The present invention relates to the fields of molecular biology, combinatorial chemistry and biochemistry. Particularly, the present invention describes methods and kits for dynamically reducing the variance between analyte taken from complex mixtures.

1 - 39 . (canceled)  
   
   
       40 . A method for reducing the relative amounts of analytes in a sample comprising: 
 a) providing a test sample comprising first different amounts of a plurality of different analytes, wherein the first different amounts have a first variance;    b) contacting the test sample with amounts of each of a plurality of different binding moieties;    c) capturing amounts of the different analytes from the test sample with the different binding moieties and removing unbound analytes; and    d) isolating the captured analytes from the binding moieties to produce a second sample comprising second amounts of a plurality of different analytes, wherein the second amounts have a second variance;    wherein the amounts of each of the plurality of different binding moieties are selected to capture amounts of the different analytes whereby the second variance is less than the first variance.    
   
   
       41 . The method of  claim 40 , wherein the test sample is a biological fluid selected from the group consisting of blood, biopsy tissue amniotic fluid, milk, urine, sweat, saliva, a cell extract or derivatives thereof.  
   
   
       42 . The method of  claim 40 , wherein the test sample is an environmental sample.  
   
   
       43 . The method of  claim 40 , wherein the plurality of different analytes is at least four different analytes.  
   
   
       44 . The method of  claim 40 , wherein the plurality of different analytes is at least one thousand, at least ten thousand, at least one-hundred thousand, at least one million or at least ten million different analytes.  
   
   
       45 . The method of  claim 40 , wherein the contacting step comprises admixing the binding moieties with the test sample.  
   
   
       46 . The method of  claim 40 , wherein the binding moieties are a combinatorial library.  
   
   
       47 . The method of  claim 40 , further comprising fractionating the captured analytes based on a physical or chemical property.  
   
   
       48 . The method of  claim 40 , wherein the plurality of binding moieties comprises bio-organic polymers.  
   
   
       49 . The method of  claim 46 , wherein the bio-organic polymers are selected from the group consisting of peptides, oligonucleotides and oligosaccharides.  
   
   
       50 . The method of  claim 49 , wherein the bio-organic polymers are peptides at least four amino acids in length.  
   
   
       51 . The method of  claim 49 , wherein the peptides are antibodies.  
   
   
       52 . The method of  claim 51 , wherein the antibodies are surface molecules of a phage display library.  
   
   
       53 . The method of  claim 49 , wherein the bio-organic polymers are oligonucleotides of at least fifteen units in length.  
   
   
       54 . The method of  claim 53 , wherein the binding moieties are aptamers.  
   
   
       55 . The method of  claim 53 , wherein the binding moieties are oligosaccharides of at least five monosaccharide units in length.  
   
   
       56 . The method of  claim 40 , wherein the contacting step is performed with the analyte in a gaseous state, a liquid state or in a suspension.  
   
   
       57 . The method of  claim 40 , wherein removing unbound analytes comprises washing the captured analytes with a wash buffer.  
   
   
       58 . The method of  claim 40 , wherein isolating the captured analytes comprises eluting the analytes from the binding moieties by contacting the captured analytes with an elution buffer and collecting the analytes in the elution buffer free of binding moieties.  
   
   
       59 . The method of  claim 58 , wherein the elution buffer is a solution comprising a component selected from the group consisting of a detergent a salt, an organic solvent, an acid, a base, a chaotropic agent and a competing ligand for the binding moiety.  
   
   
       60 . The method of  claim 40 , wherein the second variance is at least 25% less than the first variance.  
   
   
       61 . The method of  claim 40 , further comprising fractionating the analytes into subgroups.  
   
   
       62 . The method of  claim 61 , wherein fractionating comprises segregating the analytes using a technique selected from the group consisting of chromatography, electrophoresis, capillary electrophoresis, filtration and precipitation.  
   
   
       63 . The method of  claim 40 , wherein the different binding moieties are coupled to a solid support.  
   
   
       64 . The method of  claim 63 , wherein the solid support comprises a hydrogel.  
   
   
       65 . The method of  claim 63 , wherein the coupling comprises a covalent bond.  
   
   
       66 . The method of  claim 63 , wherein the solid support is a plurality of insoluble surfaces, each insoluble surface coupled to a single species of binding moiety.  
   
   
       67 . The method of  claim 66 , wherein the insoluble surfaces are beads.  
   
   
       68 . The method of  claim 63 , wherein the binding moieties are coupled to the solid support by a capture moiety.  
   
   
       69 . The method of  claim 68 , wherein the capture moiety is selected from the group consisting of FLAG tag, His tag and biotin.  
   
   
       70 . The method of  claim 40 , further comprising detecting the isolated analytes.  
   
   
       71 . The method of  claim 63 , wherein detecting the analytes is performed using an assay comprising a technique selected from the group consisting of: colorimetric, spectrophotometric, magnetic resonance, ellipsometric, mass spectroscopic, electrophoretic, chromatographic and enzymatic.  
   
   
       72 . The method of  claim 64 , wherein the assay is mass spectroscopic and the second variance of the isolated analytes allows quantifying at least 30% of the captured analytes using a single detector setting.  
   
   
       73 . A kit for detecting a plurality of analytes in a sample, comprising: 
 (a) a plurality of binding moieties, each binding moiety in an amount selected to capture a pre-determined amount of a different analyte;    (b) a plurality of containers retaining components for sample preparation and analyte isolation; and    (c) instructions for using the kit.    
   
   
       74 . The kit of  claim 73 , wherein the binding moieties are coupled to the solid support.  
   
   
       75 . The kit of  claim 73 , wherein the components for sample preparation comprise a wash solution sufficient for removing unbound material from a binding moiety specifically bound to an analyte, and an elution solution sufficient to release analyte specifically bound by a binding moiety.