Analytical biochemistry system with robotically carried bioarray

Abstract

An analytical biochemistry system featuring a substrate with reactants immobilized thereon at fixed, known locations, a holder supporting the substrate and a manipulator for transporting the holder to a fixed sample and to an inspection station. The reactants are binding agents for a target biomolecule in a sample which forms a bound substance having a detectable characteristic. The holder may be a standard pipettor, optionally carried by a robot arm or hand as the manipulator to contact the sample for detection of the presence of target biomolecules within the sample. In one embodiment, the holder is a pipette tip within which the substrate is housed, or it may be a pipette adapter which bears the substrate and fits within the sample wells of a standard microtiter plate. After appropriate incubation, the substrate and holder may be moved from contact with the sample, and the substrate may be exposed to any necessary development steps before being moved by the manipulator to a location for probing, such as by a beam. Probing of the substrate is performed to identify binding or complexing of target biomolecules of the sample with the reactants immobilized on the substrate, and may be accomplished via fluorescence detection, light scattering, autoradiography, or some other detection method.

1 . A system for detecting the presence of a target biomolecule within a sample comprising: 
 a support surface treated with distinct reactants immobilized thereon in a spaced-apart relation, forming a substrate having a plurality of spaced-apart active sites, at least one of the reactants being reactive with a target biomolecule to form a bound complex having a detectable characteristic,    holder means for supporting the substrate,    an inspection station having means for probing the spaced-apart active sites of the substrate for said detectable characteristic of the sample, and    manipulator means for bringing the holder means into contact with the sample and into said inspection station.    
     
     
         2 . The system of  claim 1  further comprising a plurality of sample reservoirs arranged in fixed, closely spaced, known locations, said manipulator means comprising a robotic arm movable among said sample holders and said inspection station.  
     
     
         3 . The system of  claim 1  wherein the holder means comprises a pipettor.  
     
     
         4 . The system of  claim 2  wherein the robotic arm comprises a movable upright tower supporting a cross beam, wherein said holder means receives cantilever support from the cross beam.  
     
     
         5 . The system of  claim 1  wherein the substrate has opposed ends fixed to the holder means and an unsupported region between the opposed ends.  
     
     
         6 . The system of  claim 5  wherein the holder means comprises a pipette tip within which the sample may be drawn by aspiration.  
     
     
         7 . The system of  claim 6  wherein the pipette tip supports the substrate longitudinally along an inside wall of the pipette tip.  
     
     
         8 . The system of  claim 6  wherein the pipette tip has an optical surface.  
     
     
         9 . The system of  claim 6  wherein the pipette tip has a narrow bore for minimizing sample volume and maximizing proximity of the sample and the substrate.  
     
     
         10 . The system of  claim 6  wherein the pipette tip has at least one flattened surface for minimizing sample volume and maximizing proximity of the sample and the substrate, and for enhancing optical inspection of the substrate.  
     
     
         11 . The system of  claim 5  wherein the holder means comprises a plunger-type pipette tip within which the sample may be drawn by withdrawal of the plunger and resulting aspiration of the sample and the sample may be dispensed by depression of the plunger.  
     
     
         12 . The system of  claim 11  wherein the plunger-type pipette tip supports the substrate longitudinally along an inside wall of the plunger-type pipette tip.  
     
     
         13 . The system of  claim 11  wherein the plunger-type pipette tip has an optical surface.  
     
     
         14 . The system of  claim 11  wherein the plunger-type pipette tip has a narrow bore for minimizing sample volume and maximizing proximity of the sample and the substrate.  
     
     
         15 . The system of  claim 1  wherein the holder means comprises a pipette adapter having a bracket for holding the substrate.  
     
     
         16 . The system of  claim 15  wherein the pipette adapter has a coupler for fitting the pipette adapter to a pipetting tool.  
     
     
         17 . The system of  claim 15  wherein the pipette adapter is sufficiently narrow at the bracket to immerse the bioarray in a sample well of a microtiter plate.  
     
     
         18 . The system of  claim 15  wherein the bracket of the pipette adapter holds the substrate between opposed prongs of the bracket.  
     
     
         19 . The system of  claim 15  wherein the substrate is planar and the pipette adapter supports the substrate in an orientation such that the active sites face downward.  
     
     
         20 . The system of  claim 15  further comprising knobs on the bracket prongs, the knobs positioning the substrate a fixed distance from the sample in a position to protect the substrate from abrasion and contamination.  
     
     
         21 . The system of  claim 15  further comprising an evaporation barrier disposed about the pipette adapter.  
     
     
         22 . The system of  claim 1  wherein the manipulator means comprises a pipette adapter having a flat bottom surface for bearing the substrate.  
     
     
         23 . The system of  claim 22  wherein the pipette adapter has a coupler means for fitting the pipette adapter to a pipetting tool.  
     
     
         24 . The system of  claim 22  wherein the pipette adapter is sufficiently narrow at the flat bottom surface to immerse the substrate in a sample well of a microtiter plate.  
     
     
         25 . The system of  claim 22  wherein the substrate is attached to the flat bottom surface.  
     
     
         26 . The system of  claim 22  wherein the substrate is planar and the pipette adapter supports the substrate in an orientation such that the active sites face downward.  
     
     
         27 . The system of  claim 22  further comprising knobs on the flat bottom surface of the pipette adapter, the knobs positioning the substrate a fixed distance from the sample in a position to protect the substrate from abrasion and contamination.  
     
     
         28 . The system of  claim 22  further comprising an evaporation barrier disposed about the flat-bottom pipette adapter.  
     
     
         29 . The system of  claim 1  wherein the substrate is a flat bottom surface of a pipette adapter.  
     
     
         30 . The system of  claim 1  wherein the substrate has a linear arrangement of the reactants.  
     
     
         31 . The system of  claim 30  wherein the substrate has a segmented linear arrangement of the reactants.  
     
     
         32 . The system of  claim 1  wherein the substrate has an area-wide arrangement of the reactants.  
     
     
         33 . The system of  claim 1  wherein the substrate is preformed with reactants prior to being affixed to the manipulator means.  
     
     
         34 . The system of  claim 1  wherein the substrate is affixed to the manipulator means before reactants are immobilized on the substrate.  
     
     
         35 . The system of  claim 1  wherein the manipulator means has a surface and the substrate is formed directly on the manipulator means by utilizing the surface as the substrate.  
     
     
         36 . The system of  claim 1  wherein the reactants comprise complementary DNA strands.  
     
     
         37 . The system of  claim 1  wherein the reactants comprise immunological biomolecules.  
     
     
         38 . The system of  claim 1  wherein the substrate comprises a polypropylene member.  
     
     
         39 . The system of  claim 1  wherein the substrate comprises a glass member.  
     
     
         40 . The system of  claim 1  wherein the reactants are anchored on the substrate via biopolymer synthesis.  
     
     
         41 . The system of  claim 1  wherein the reactants are anchored on a solid support member via biopolymer synthesis with subsequent segmenting of the solid support member and mounting of the solid support member on the support surface to form said substrate.  
     
     
         42 . The system of  claim 1  wherein the substrate has reactants deposited at discrete known locations on the substrate.  
     
     
         43 . The system of  claim 42  wherein reactants are dispensed onto the substrate through movable nozzles.  
     
     
         44 . The system of  claim 1  wherein the reactants are affixed to the substrate via covalent linkage.  
     
     
         45 . The system of  claim 1  wherein the reactants are affixed to the substrate via physical absorption.  
     
     
         46 . The system of  claim 1  wherein the inspection station further comprises: 
 means for impinging the light beam on the substrate,  
 a light collector for collecting optical signals emitted from the substrate, and  
 a detector disposed to detect the collected signals from the light collector.  
 
     
     
         47 . The system of  claim 46  wherein the detector comprises a fluorescence detector.  
     
     
         48 . The system of  claim 1  wherein the inspection station comprises a beam passing through the substrate to a detector measuring absorbance.  
     
     
         49 . The system of  claim 1  wherein the inspection station comprises a beam impinging on the substrate with a detector disposed for measuring reflectance.  
     
     
         50 . The system of  claim 1  wherein the inspection station comprises a beam impinging on the substrate with a detector disposed for measuring light scattering.  
     
     
         51 . The system of  claim 1  wherein the inspection station comprises a beam impinging on the substrate with a detector disposed for measuring chemiluminescence.  
     
     
         52 . A method for detecting the presence of a target biomolecule within a sample, the method comprising: 
 (a) immobilizing distinct reactants in a spaced-apart relation on a surface of a support member, thereby forming a substrate having a plurality of active sites, at least one of the reactants being reactive with a target biomolecule to form a bound substance having a detectable characteristic,    (b) affixing the substrate onto a holder,    (c) moving the holder into contact with the sample for a period of time,    (d) removing unbound biomolecules from the substrate, and    (e) moving the substrates to a location for probing the active sites of the substrate, and    (f) detecting a detectable characteristic of the bound structure on the substrate.    
     
     
         53 . The method of  claim 52  further defined by robotically moving the holder.  
     
     
         54 . The method of  claim 52  wherein immobilizing the reactants comprises 
 synthesizing reactants by biopolymerization reactions anchored at discrete locations on the substrate.  
 
     
     
         55 . The method of  claim 52  wherein immobilizing the reactants further comprises 
 synthesizing reactants by biopolymerization reactions anchored at discrete locations on a solid support surface,  
 segmenting the solid support surface to separate distinct reactants, and  
 mounting the segmented solid support surface to the support member to form said substrate with spaced-apart active sites.  
 
     
     
         56 . The method of  claim 52  wherein immobilizing the reactants further comprises 
 dispensing the reactants at discrete locations on the substrate via movable nozzles.  
 
     
     
         57 . The method of  claim 56  further comprising 
 activating the surface of the support member before dispensing the reactants.  
 
     
     
         58 . The method of  claim 56  further comprising 
 affixing an optical bar code pattern to the support member for identification of the substrate.  
 
     
     
         59 . The method of  claim 56  wherein dispensing the reactants at discrete locations further comprises directing the reactants to form round spots on the support member.  
     
     
         60 . The method of  claim 56  wherein dispensing the reactants at discrete locations further comprises directing the reactants to form linear bands on the support member.  
     
     
         61 . The method of  claim 56  wherein dispensing the reactants at discrete locations further comprises deflecting the reactants to form a desired pattern on the support member.  
     
     
         62 . The method of  claim 52  wherein immobilizing the reactants further comprises 
 affixing excess amount of reactants to the support member compared to the amount of suspected target biomolecules of the sample which would react with the reactants.  
 
     
     
         63 . The method of  claim 52  wherein immobilizing the reactants comprises forming covalent linkages of the reactants with the support member.  
     
     
         64 . The method of  claim 52  wherein immobilizing the reactants comprises adsorption of the reactants on the support member.  
     
     
         65 . The method of  claim 52  wherein affixing the substrate onto a holder further comprises affixing the substrate in a linear arrangement along an inside wall of a pipette tip, and wherein contacting the sample within the substrate further comprises aspirating the sample into the pipette tip.  
     
     
         66 . The method of  claim 52  wherein affixing the substrate onto a holder further comprises affixing the substrate in a linear arrangement along an inside wall of a plunger-type pipette tip, wherein contacting the sample with the substrate further comprises drawing the sample into a plunger-type pipette tip by withdrawal of the plunger, and wherein removal of the substrate from the sample further comprises dispensing the sample from the plunger-type pipette tip by depression of the plunger.  
     
     
         67 . The method of  claim 52  wherein affixing the substrate onto a holder further comprises affixing the substrate between opposed prongs of a bracket of a pipette adapter, and wherein contacting the sample with the substrate further comprises placing the sample in a vial or well and placing the pipette adapter, with the prongs in an inverted position, into the vial or well.  
     
     
         68 . The method of  claim 52  wherein affixing the substrate onto a holder further comprises affixing the substrate to a flat, bottom surface of a pipette adapter and wherein contacting the sample with the substrate further comprises placing the sample in a vial or well and placing the pipette adapter, with the flat, bottom surface down, into the vial or well.  
     
     
         69 . The method of  claim 52  further comprising 
 exposing the substrate to a development chemical to enhance visualization of complementation, after removing the substrate from the sample and before probing the active site.  
 
     
     
         70 . The method of  claim 69  wherein exposing the substrate to a development chemical further comprises exposing the substrate to a fluorescence dye.  
     
     
         71 . The method of  claim 52  wherein detecting a detectable characteristic comprises inspecting the bound structure with a light beam for fluorescence.  
     
     
         72 . The method of  claim 52  wherein detecting a detectable characteristic comprises inspecting the bound structure with a light beam for absorbance.  
     
     
         73 . The method of  claim 52  wherein detecting a detectable characteristic comprises inspecting the bound structure with a light beam for reflectance.  
     
     
         74 . The method of  claim 52  wherein detecting a detectable characteristic comprises inspecting the bound structure with a light beam for light scattering.  
     
     
         75 . The method of  claim 52  wherein detecting a detectable characteristic comprises inspecting the bound structure for chemiluminescence.  
     
     
         76 . The method of  claim 52  further comprising 
 recording the sample volume exposed to the substrate,  
 quantifying the amount of bound substance at the active sites on the substrate, and  
 calculating the amount of the target biomolecule present in the sample from the sample volume exposed to the substrate and the amount of bound substance at the active sites of the substrate.  
 
     
     
         77 . A holder for bearing a substrate having reactant binding agents for at least one target biomolecule in a sample to form a bound substance having a detectable characteristic for detection of target biomolecules within the sample, the holder comprising: 
 a pipette tip for use in conjunction with a pipettor, the pipette tip bearing a substrate with distinct reactants in a spaced-apart relation, the substrate disposed longitudinally along an inside wall of the pipette tip,    wherein the pipette tip has means for aspirating the sample into the pipette tip for contact between the reactants and the sample on the substrate.    
     
     
         78 . The holder of  claim 77  wherein the aspirating means is a robotic pipettor.  
     
     
         79 . The holder of  claim 77  wherein the aspirating means is a hand-held pipettor.  
     
     
         80 . The holder of  claim 77  wherein the pipette tip has a narrow bore to maximize proximity of the substrate and the sample.  
     
     
         81 . The holder of  claim 77  wherein the pipette tip has at least one flattened surface.  
     
     
         82 . The holder of  claim 77  wherein the pipette tip has an optical surface opposite the substrate.  
     
     
         83 . A holder for bearing a substrate having reactant binding agents for at least one target biomolecule in a sample to form a bound substance having a detectable characteristic for detection of target biomolecules within the sample, the holder comprising 
 a pipette tip having a plunger and bearing a substrate with distinct reactants in a spaced-apart relation, the substrate disposed longitudinally along an inside wall of the pipette tip,    wherein the plunger draws the sample into the pipette tip for contact between the reactants and the sample on the substrate.    
     
     
         84 . The holder of  claim 83  wherein withdrawal of the plunger is automated.  
     
     
         85 . The holder of  claim 83  wherein the pipette tip has a narrow bore to maximize proximity of the substrate and the sample.  
     
     
         86 . The holder of  claim 83  wherein the pipette tip has at least one flattened surface.  
     
     
         87 . The holder of  claim 83  wherein the pipette tip has an optical surface opposite the substrate.  
     
     
         88 . A holder for bearing a substrate having reactant binding agents for at least one target biomolecule in a sample to form a bound substance having a detectable characteristic for detection of target biomolecules within the sample, the holder comprising: 
 a pipette adapter having a bracket with opposed prongs, the opposed prongs bearing the substrate with distinct reactants in a spaced-apart relation, the substrate disposed in a downward-facing orientation,    wherein the bracket of the pipette adapter is placed on the sample for contact of the sample with the substrate.    
     
     
         89 . The holder of  claim 88  wherein the pipette adapter has a coupling means for coupling the pipette adapter to a robotic arm.  
     
     
         90 . The holder of  claim 88  wherein the pipette adapter has a coupling means for coupling the pipette adapter to a hand-held pipette.  
     
     
         91 . The holder of  claim 88  wherein the bracket of the pipette adapter is sufficiently narrow in diameter to fit within a sample well of a microtiter plate.  
     
     
         92 . The holder of  claim 88  wherein the bracket prongs of the pipette adapter have knobs positioning the substrate a fixed distance from the sample.  
     
     
         93 . The holder of  claim 88  wherein the pipette adapter has an evaporation barrier disposed about the pipette adapter.  
     
     
         94 . A holder for bearing a substrate having reactant binding agents for at least one target biomolecule in a sample to form a bound substance having a detectable characteristic for detection of target biomolecules within the sample, the holder comprising: 
 a pipette adapter having an end with a flat bottom surface, the flat bottom surface bearing the substrate with distinct reactants in a spaced-apart relation, the substrate disposed in a downward facing orientation,    wherein the flat bottom surface of the pipette adapter is placed on the sample for contact of the sample with the substrate.    
     
     
         95 . The holder of  claim 94  wherein the pipette adapter has a coupling means for coupling the pipette adapter to a robotic arm.  
     
     
         96 . The holder of  claim 94  wherein the pipette adapter has a coupling means for coupling the pipette adapter to a hand-held pipettor.  
     
     
         97 . The holder of  claim 94  wherein the end of the pipette adapter having the flat bottom surface is sufficiently narrow in diameter to fit within a sample well of a microtiter plate.  
     
     
         98 . The holder of  claim 94  wherein the flat bottom surface of the pipette adapter has knobs positioning the substrate a fixed distance from the sample.  
     
     
         99 . The holder of  claim 94  wherein the pipette adapter has an evaporation barrier disposed about the pipette adapter.