Methods and apparatus for conducting multiple measurements on a sample

Abstract

Multiplexed test measurements are conducted using an assay module having a plurality of assay domains. In preferred embodiments, these measurements are conducted in assay modules having integrated electrodes with a reader apparatus adapted to receive assay modules, induce luminescence, preferably electrode induced luminescence, in the wells or assay regions of the assay modules and measure the induced luminescence.

1 . An assay module for conducting luminescence assays comprising a plurality of assay domains, said assay domains comprising a first assay domain having a first reagent and a second assay domain having a second reagent, wherein said first assay domain is capable of generating luminescence at least 10 times brighter than said second assay domain and said first assay domain and said second assay domains are not adjacent and/or are separated by at least one other assay domain to reduce luminescence emitted from said first assay domain from interfering with luminescence emitted from said second assay domain. 
     
     
         2 . The assay module of  claim 1 , wherein said first assay domain comprises at least 10 times capture reagent than said second assay domain. 
     
     
         3 . The assay module of  claim 1 , wherein said first assay domain is capable of generating luminescence at least 100 times brighter than said second assay domain. 
     
     
         4 . The assay module of  claim 1 , wherein said first assay domain is capable of generating luminescence at least 1,000 times brighter than said second assay domain. 
     
     
         5 . The assay module of  claim 1 , wherein said second assay domain has one or more properties that diminish emitted luminescence. 
     
     
         6 . The assay module of  claim 5 , wherein said one or more properties include a capture reagent that diminishes emitted luminescence and/or a target analyte that diminishes emitted luminescence. 
     
     
         7 . The assay module of  claim 1 , wherein said first assay domain is separated from said second assay domain by at least one other assay domain. 
     
     
         8 . The assay module of  claim 1 , wherein said first assay domain and said second assay domain are separately addressable. 
     
     
         9 . The assay module of  claim 1 , wherein said assay module is a multi-well plate comprising a plurality of wells, each of said wells having a copy of said plurality of assay domains. 
     
     
         10 . An assay module comprising a plurality of assay domains for measuring a panel of analytes, said panel comprising a plurality of analytes that are members of an analyte category selected from cytokines and/or their receptors; growth factors and/or their receptors; second messengers; drugs of abuse; therapeutic drugs; auto-antibodies; allergen specific antibodies; tumor markers; cardiac markers; markers associated with hemostasis; markers of acute viral hepatitis infection; markers of Alzheimers Disease; markers of osteoporosis; markers of fertility; markers of congestive heart failure; markers of thyroid disorders; markers of prostrate cancer; nucleic acid arrays for measuring mRNA levels of mRNA coding for cytokines, growth factors, components of the apoptosis pathway, expression of the P450 enzymes or expression of tumor related genes; nucleic acid arrays for genotyping individuals, pathogens, or tumor cells. 
     
     
         11 . An assay module of  claim 10 , wherein said assay domains comprise: (i) analytes or analogs of analytes; (ii) binding reagents that bind analytes; and/or (iii) substrates for catalytic activity of analytes. 
     
     
         12 . A method of performing an assay for a plurality of analytes using an assay module of  claim 10 , said method comprising:
 (a) contacting a sample with a plurality of assay domains within said assay module; and   (b) measuring said analytes at said assay domains.   
     
     
         13 . A method for detecting or correcting for a source of assay error in an assay for an analyte comprising:
 (a) introducing a sample comprising analyte into an assay module comprising   (i) a first assay domain comprising a first reagent capable of binding said analyte; and   (ii) a second assay domain comprising said first reagent;   wherein said first assay domain and said second binding domain having different properties, said different properties selected from the group consisting of surface area, reagent concentration, heat stability, pH stability, hydrophobic or hydrophilic properties, co-reactants, epitope specificity, binding kinetics, non-specific binding properties, binding constants, and/or thermodynamics; and   (b) comparing the binding of the analyte to the first and second assay domains so as to detect or correct for sources of assay error.   
     
     
         14 . An assay module for conducting an assay on a sample comprising an analyte and at least one interferent, said module comprising:
 (a) an assay domain capable of binding said analyte; and   (b) a sequestering domain capable of binding said interferent.   
     
     
         15 . The assay module of  claim 14 , wherein said module is adapted to measure said interferent bound to said sequestering domain. 
     
     
         16 . A method for measuring an analyte in the presence of an interferent comprising:
 (a) introducing the sample into the assay module of  claim 14 ; and   (b) detecting the binding of said analyte to said assay domain.   
     
     
         17 . An assay module comprising a plurality of assay domains, wherein said plurality of assay domains comprises one or more assay domains comprising one or more binding reagents and further comprising a first control domain comprising pre-bound label, a second control domain providing a defined reaction and a third control domain consisting essentially of blocking agent. 
     
     
         18 . The assay module of  claim 17 , wherein said first control domain controls for one or more of the following: (i) chemical interference with ECL generation; (ii) optical interference with light transmission; and/or (iii) module variation. 
     
     
         19 . The assay module of  claim 17 , wherein said second control domain controls for one or more of the following: (i) non-specific binding or biochemical activity; (ii) chemical interference with ECL generation; (iii) optical interference with light transmission; (iv) pipetting errors; (v) timing errors; (vi) variations in mixing; (vii) variations in temperature; and (viii) assay module variation. 
     
     
         20 . The assay module of  claim 17 , wherein said third control domain controls for non-specific binding. 
     
     
         21 . An assay module comprising one or more assay domains comprising assay reagents and one or more sequestrating domains comprising sequestration agents for sequestering interfering species. 
     
     
         22 . The assay module of  claim 21 , wherein said one or more sequestrating domains sequest one or more of the following interfering species: biotin; anti-Streptavidin; hemoglobin; bilirubin; lipid; high and low albumin; HAMA; anti-ruthenium; high Rheumatoid Factor; DLIF's for digoxin or digitoxin type molecules; drugs/co meds; cross reactive analytes; and combinations thereof. 
     
     
         23 . An assay module comprising one or more assay domains comprising assay reagents and one or more sequestrating domains comprising sequestration agents capable of sequestering a labeled reagent.