Broad specificity affinity arrays: a qualitative approach to complex sample discrimination

Abstract

Described is a method for discriminating complex biological samples using an array of discrete biological sensing elements immobilized onto a solid support in which constituents bound to the sensor array is directly determined by measuring the mass increase on the surface; data analysis of said method is performed using neutral network or statical based pattern recognition techniques. In a preferred embodiment the liquid sample is tested for the presence of soluble constituent(s) by contacting said sample with said sensor array under specific conditions, removing unbound sample constituent(s), determining the mass increase on the surface and comprising said mass increase data with a reference standard using pattern recognition software.

1 . A method for discriminating complex biological samples using an array of discrete biological sensing elements immobilized onto a solid support inwhich constituents bound to the sensor array is directly determined by measuring the mass increase on the surface; data analysis of said method is performed using neural network or statical based pattern recognition techniques. 
     
     
         2 . A method for discriminating complex biological samples using an array of discrete biological sensing elements immobilized onto a solid support and testing the liquid sample for the presence of soluble constituent(s) comprising: contacting said sample with said sensor array under conditions wherein said sensing elements permitting binding of constituents in said sample; if any are present; removal of substantially all unbound sample constituents; the direct detection of said bound constituents by determining the mass increase of said components on the surface; comparison of said mass increase data of said sample with a reference standard using pattern recognition software. 
     
     
         3 . The method of  claim 2  wherein said sensing element is selected from a group consisting of, antibodies, lectins, nucleic acids, carbohydrates, genetically modified derivatives and combinations and gradients thereof. 
     
     
         4 . The method in  claim 2  wherein said sensing element is of nonbiological origin but is endowed with biological-like recognition selected from the group consisting of cyclodextran and derivatives thereof, roxane and derivatives thereof, templated or imprinted polymers and combinations thereof. 
     
     
         5 . The method in any of  claims 2  to  4  wherein said immobilization procedure combines the use of gold surfaces, long chain thiol alkanes with terminal activatable groups, hydrophobic/hydrophilic patterning, EDC/NHS coupling amino-biotin and streptavidin. 
     
     
         6 . The method in any of  claims 2  to  5  wherein said solid support is selected from the group consisting of silicon, glass, mica, plastic, platinum, silver, copper, gold and combinations thereof. 
     
     
         7 . The method in any of  claims 2  to  6  wherein said surface mass increase detection techniques are selected from the group consisting of quartz crystal microbalances, optoaucostics, reflectometry, ellipsometry, SAW and surface plasmon resonance. 
     
     
         8 . The method in any of  claims 2  to  7  wherein said surface mass detection is performed in imaging mode with a CCD camera. 
     
     
         9 . The method of diagnosing a disease, said method comprising the method in any of  claims 2  to  8 , wherein said mass increase pattern is diagnostic of the disease, said sample is a patient sample and said standard is the pattern present in an individual without said disease. 
     
     
         10 . The method in  claim 9  wherein said patient sample is a human or animal, tissue or bodily fluid, selected from the group consisting of blood, serum, urine, milk, sweat, exhaled air, skin, bone marrow, cerebrospinal fluid, synovial fluid, amniotic fluid and lymphatic fluid. 
     
     
         11 . The method in  claim 9  wherein said disease is selected from the group consisting of genetic disorders, autoimmune diseases, arthritis, infectious diseases, cancer, heart disease, drug abuse and lung disease. 
     
     
         12 . The method of diagnosing the general state of health, said method comprising the method in any of  claims 2  to  11 , wherein said mass increase pattern is diagnostic of a particular state of health, said sample is a patient sample and said standard is the pattern present in a representative part of the population. 
     
     
         13 . The method in  claim 12  wherein said general state of health is selected from common mild ailments and/or health conditions with diffuse symptoms, consisting of high blood pressure, pregnancy, common colds, injuries, inflammatory reactions, mild immune suppression, doping, altitude sickness, space sickness chronic fatigue syndrome; and effects of low level toxic chemical or radiation exposure, menstrual cycles and subclinical infections. 
     
     
         14 . The method of identifying an organism, said method comprising the method in any of  claims 2  to  8 , wherein said mass increase pattern is unique to a particular organism, said sample is a biological sample from a particular organism and said standard is the pattern normally found in that organism. 
     
     
         15 . The method in  claim 14  wherein said biological sample is tissue or an extract selected from the group consisting of animals, microorganisms, fungi, viruses, bacteria, plants and protozoa. 
     
     
         16 . The method of identifying samples contaminated with toxic compounds, said method comprising the method in any of  claims 2  to  8 , wherein said mass increase pattern is diagnostic of contaminated material, said sample is a environment sample and said standard is the pattern present in an uncontaminated sample. 
     
     
         17 . The method in  claim 16  wherein said environmental sample is the untreated or extracted sample selected from the group consisting of air, soil, water, rock, ice, plant, lichen, animal and food stuffs. 
     
     
         18 . The method in any of  claims 2  to  17  wherein said sensing element array are lectins selected from the group consisting of canavalia ensiformis, bandeiraea simplicifolia BS-I, arachis hypogaea, phytolacca americana, phaseolus vulgaris pha-e, artocarpus integrifolia, triticum vulgaris, pisum sativum. 
     
     
         19 . The method in any of  claims 2  to  18  wherein said the lectin array is expanded to include additional lectin or lectin-like sensing elements and/or used in combination with other biological sensing elements in a diagnostic assay. 
     
     
         20 . The method in any of  claims 2  to  8  wherein said sensing elements are used in gradients in pure and mixed formats and in any combination thereof. 
     
     
         21 . The method in any of  claims 2  to  18  wherein said bound constituents are isolated and used as diagnostic tool, to further the understanding of the disease or to develop a drug or therapy to treat an ailment identified by the said method.