Simultaneous particle separation and chemical reaction
First Claim
1. A method for reacting small primary particles from a primary stream also comprising larger particles, comprising the steps of:
- conducting said primary stream into a laminar flow reaction channel;
separately conducting a reagent stream comprising reagent particles into said reaction channel, such that said primary stream and said reagent stream flow in adjacent laminar streams;
allowing said primary particles to diffuse from said primary stream into said reagent stream, and to react with said reagent particles and form detectable product particles, thereby converting said reagent stream into a product stream and said primary stream into a residual primary stream;
conducting said residual primary stream out of said reaction channel;
separately conducting said product stream out of said reaction channel; and
detecting said product particles.
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Abstract
This invention provides a method and apparatus for detecting the presence of analyte particles in a sample fluid also comprising larger particles, particularly blood. It exploits diffusion to provide simultaneous filtering of the larger particles and reaction of the analyte particles. A sample stream and a reagent stream join on the upstream end of a laminar flow reaction channel and flow in adjacent laminar streams. The reagents can be in solution or immobilized on a bead. The analyte particles diffuse from the sample stream into the reagent stream, leaving behind the larger particles in the residual sample stream. In the reagent stream the analyte particles react with reagent particles and form product particles, thereby creating a product stream. At the downstream end of the reaction channel, the residual sample stream and the product stream are divided. The product particles are then detected, preferably optically, in the product stream. Prior to detection, the product stream can undergo further filtering or separation, or can join a second reagent stream to form secondary product particles. This apparatus and method can be used to implement competitive immunoassays or sandwich immunoassays using enzymatically or fluorescently labeled immunoreagents. The apparatus and method can also be used to synthesize products, in which case two reagent streams join in the laminar flow reaction channel.
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Citations
21 Claims
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1. A method for reacting small primary particles from a primary stream also comprising larger particles, comprising the steps of:
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conducting said primary stream into a laminar flow reaction channel;
separately conducting a reagent stream comprising reagent particles into said reaction channel, such that said primary stream and said reagent stream flow in adjacent laminar streams;
allowing said primary particles to diffuse from said primary stream into said reagent stream, and to react with said reagent particles and form detectable product particles, thereby converting said reagent stream into a product stream and said primary stream into a residual primary stream;
conducting said residual primary stream out of said reaction channel;
separately conducting said product stream out of said reaction channel; and
detecting said product particles. - View Dependent Claims (2, 3)
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4. A method for reacting primary particles from a primary stream, comprising the steps of:
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conducting said primary stream into a first laminar flow reaction channel;
separately conducting a first reagent stream comprising first reagent particles into said first laminar flow reaction channel, such that said primary stream and said first reagent stream flow in adjacent laminar streams;
allowing said primary particles to diffuse from said primary stream into said first reagent stream, and to react with said first reagent particles and form first product particles, thereby converting said first reagent stream into a first product stream and said primary stream into a residual primary stream;
thereafter conducting a first companion stream into said first laminar flow reaction channel such that said first product stream and said first companion stream flow in adjacent laminar streams thereby converting said first product stream into a diffused first product stream and said first companion stream into a diffused first companion stream;
conducting said residual primary stream out of said first reaction channel; and
separately conducting said first diffused product stream out of said first reaction channel. - View Dependent Claims (5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method for detecting antigens in whole blood comprising the steps of:
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conducting a primary stream comprising whole blood containing first antigens into a laminar flow reaction channel;
separately conducting into said reaction channel a reagent stream comprising fluorescently-labeled second antigens identical to said first antigens, complexed with antibodies to said antigens, to form a complex, such that said primary stream and said reagent stream flow in adjacent laminar streams in said channel;
allowing first antigens from said blood to diffuse from said primary stream into said reagent stream, and to react with said complex to displace fluorescently-labeled second antigens therefrom, thereby converting said reagent stream into a product stream and said primary stream into a residual stream;
conducting said residual stream out of said reaction channel;
separately conducting said product stream out of said reaction channel; and
detecting displaced fluorescently-labeled second antigens in said product stream. - View Dependent Claims (15, 16)
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17. A method for detecting antigens in whole blood comprising the steps of:
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conducting a primary stream comprising whole blood containing first antigens into a laminar flow reaction channel;
separately conducting into said reaction channel a reagent stream comprising beads having attached thereto fluorescently-labeled second antigens, identical to said first antigens, complexed with antibodies to said antigens to form a complex, such that said primary stream and said reagent stream flow in adjacent laminar streams;
allowing first antigens from said blood to diffuse from said primary stream into said reagent stream and to react with said complex to displace fluorescently-labeled second antigens on said beads;
thereafter detecting fluorescently-labeled beads and/or fluorescently-labeled second antigens. - View Dependent Claims (18, 19, 20, 21)
conducting said residual stream out of said reaction channel; and
separately conducting said product stream out of said reaction channel via a detection channel.
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21. The method of claim 20 wherein said detection channel is convoluted.
Specification