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All Rights Reserved AEIC 2004
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Protein Au Vial Absorbent Pad Test Line Sample Extract Filter Pad Gold Pad Membrane Control Line
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Step 6 Data analysis Sample plant tissue Step 1 Weigh Sample Aliquots Step 2 Extract Samples Step 3 Aliquot extracts for analysis and sample back-ups Step 4 ELISA Analysis Step 5
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All Rights Reserved AEIC 2004
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0 1 2 3 4 0.010.1110100 % GMO Corn OD Event176 Bt11 Mon810
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All Rights Reserved AEIC 2004 y = - 0.1567 x 2 + 1.3795x + 0.0656 2 R = 0.9999 0.0 0.5 1.0 1.5 2.0 2.5 3.0 0.00.51.01.52.02.5 %GMO OD 0.1% LOD Soymeal was toasted for 60 min. at 100 ºC Antibody not selective for toasted soymeal Antibody detects toasted soymeal
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Trait Agdia EnviroLogix Neogen SDI CP4 EPSPS E,S S E 1,S Cry1Ab E,S E,S E,S E 2,S Cry9C E 3,S E,S E 3,S Cry2A E,S Cry3A E NK603 S S S www.agdia.com www.envirologix.com www.neogen.comwww.sdix.com www.agdia.comwww.envirologix.comwww.neogen.comwww.sdix.com E ELISA S strip 1 validated by JRC ring test for Roundup Ready Soy bean 2 validated by AACC ring test for YieldGard (Mon810) Corn 3 official method AACC international collaborative trial All Rights Reserved AEIC 2004 ® ®
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Binding Sites Heavy Chain Light Chain
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All Rights Reserved AEIC 2004 Binding sites Proteins Heavy chain Light chain
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Target Analyte Linker Derivative Carrier Protein Immunogen All Rights Reserved AEIC 2004
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Target Analyte Analyte bound by antibody How to detect binding? All Rights Reserved AEIC 2004
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Immunoassay Conjugate
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Target Analyte Assay Conjugate Capture Antibody Detector and Analyte compete to bind with antibody All Rights Reserved AEIC 2004
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I. No Analyte - high detection signal II. Analyte present - detection signal reduced All Rights Reserved AEIC 2004
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Competitive Immunoassay Data 0 10 20 30 40 50 60 70 80 90 100 0.11101001000 Concentration of Analyte Percent of Maximum Absorbance
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Target Analyte Capture Antibody Detector Antibody Signal Recognition Sites (Epitopes) All Rights Reserved AEIC 2004
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Target proteins from the sample bind to the container walls Specific antibodies bind to their target protein and react causing a color change Photometric reader quantifies by degree of color change
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All Rights Reserved AEIC 2004 Color Substrate Bt Cry1Ab Antibodies Enzyme Bt
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All Rights Reserved AEIC 2004
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