2. All Rights Reserved AEIC 2004

15. Protein Au Vial Absorbent Pad Test Line Sample Extract Filter Pad Gold Pad Membrane Control Line

16. All Rights Reserved AEIC 2004

19. 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

20. All Rights Reserved AEIC 2004

27. 0 1 2 3 4 0.010.1110100 % GMO Corn OD Event176 Bt11 Mon810

28. 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

29. 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 ® ®

35. Binding Sites Heavy Chain Light Chain

36. All Rights Reserved AEIC 2004 Binding sites Proteins Heavy chain Light chain

37. All Rights Reserved AEIC 2004

43. Target Analyte Linker Derivative Carrier Protein Immunogen All Rights Reserved AEIC 2004

44. Target Analyte Analyte bound by antibody How to detect binding? All Rights Reserved AEIC 2004

45. Immunoassay Conjugate

46. Target Analyte Assay Conjugate Capture Antibody Detector and Analyte compete to bind with antibody All Rights Reserved AEIC 2004

47. I. No Analyte - high detection signal II. Analyte present - detection signal reduced All Rights Reserved AEIC 2004

48. Competitive Immunoassay Data 0 10 20 30 40 50 60 70 80 90 100 0.11101001000 Concentration of Analyte Percent of Maximum Absorbance

49. Target Analyte Capture Antibody Detector Antibody Signal Recognition Sites (Epitopes) All Rights Reserved AEIC 2004

50. 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

51. All Rights Reserved AEIC 2004 Color Substrate Bt Cry1Ab Antibodies Enzyme Bt

52. All Rights Reserved AEIC 2004