TETRAD Product Presentation Resources Set 1 Version 1.0 20.Oct.06

1. TETRAD Product Presentation ResourcesSet 1Version 1.020.Oct.06 CyberDiag Product Group PLN (PolyLBLnanofilm) + ChemCheck (PLN-based sensors and analyzers – chem, bio)

2. PLN / ChemCheck Slides About the PLN (PolyLBLnanofilm) core technology and its implementation in ChemCheck – for use in presentations

3. PLN Slides

4. PLN = Polymer LBL Nanofilm(aka PLC, Polymer Laminate Catalyst)[Polymer Layer-by-Layer Nanofilm Catalyst](LBL = Layer-by-Layer)

5. INTERPOLYELECTROLYTE COMPLEXES AT WATER/SOLID INTERFACE solid support PC layer-by-layer (LBL) polyelectrolyte film H2O H2O PA

6. Scheme of Layer-by-Layer Assembly by Alternate Adsorption of Oppositely Charged Linear Polyions and Nanoparticles or Proteins

7. 40-nm silica alternated with PDDA in 6 bilayer film (210 nm). (12-nm SnO2 / PDDA)18, thickness 190 nm (PAH/glucose oxidase)18 multilayer Y. Lvov, K. Ariga, T. Kunitake, Langmuir, 1997, v.13, 6195-6203 "Alternate assembly of ordered multilayers of SiO2 and other nanoparticles and polyions"

8. LATEXES BILAYER VESICLES LINEAR POLYELECTROLYTES POLYMER MICRO AND NANO GELS OBJECTS TO BE INCLUDED IN LBL FILMS METAL SOLS CARBON NANOTUBES SEMICONDUCTOR NANOPARTICLES ENZYMES AND ANTIBODIES AND OTHER CHARGED MICRO AND NANO STRUCTURES

9. Main tasks • LBL process optimization for graphite surfaces (electrodes, HOPG) • Development of biosensor electrodes • Immobilization of antibodies on the HOPG

10. Polymers PDDACl Polydimethyl-dialylammonium chloride PAS Polyanetol sulfonate Na Chitozane Poly-(1,4)-2-amino -2-deoxy-D- glucopuranose PDMEAMMABr Polydimethylethylammoniummethylmethacrylate bromide P4VP Alkyl derivatives of poly-4-vinylpyridine P-n-VBCl-Py Poly-п-vinyl-benzene chloride-pyridine

11. LBL film structures 1 2 3 4 0,05% PDDA, Images 2,5 х 2,5 mkm Adsorption from water (1), 50 mM PB (2), 50 mM PB and 0.1M NaCl (3), 50 mM PB and 1M NaCl (4)

12. LBL 4 layers

13. _ 2e PHENOL TYROSINASE BIOSENSOR Phenol 1/2 O2 Tyrosinase A principle of analysis H2O 1/2 O2 Catechol Graphite electrode Tyrosinase H2O o-Quinone

14. Choline Oxidase biosensor Choline +2O2 + H2O  betaine + 2H2O2 Mn2O G R А P H I T E Choline Oxidase PC

15. Polymer optimization for tyrosinase biosensor

16. Polymer optimization for choline oxidase biosensor

17. Adsorption Kinetic For polycation For Tyrosinase

18. Adsorption kinetic

19. Adsorption kinetic AFM images for PDDA based films PDDA, 10 min PDDA/E, 10 min

20. Number of the layers

21. Number of the layers (PC/Е)1 (PC/Е)2

22. Pretreatment Polydiethylaminophosphazen (PP)

23. Polydiethylaminophosphazen

24. Number of the layers after pretreatment

25. Tyrosinase LBL biosensor Typical response

26. Analytical parameters for tyrosinase biosensor • Linear range for phenol detection: 1∙10-8 М – 1∙10-5 М • Detection limit for phenol detection: 6 nМ • Sensitivity: 2,9 А/(М∙sm2)

27. Choline Oxidase biosensor Choline +2O2 + H2O  betaine + 2H2O2 Mn2O G R А P H I T E Choline Oxidase PC

28. Number of the layers

29. Bienzyme electrode Mn2O ChO G R А P H I T E BuChE Polycation

30. Bienzyme electrode Mn2O BuChE PC ChO G R А P H I T E 8 bilayers Detection Limit for OP – 10-11 M

31. Next-Generation PLN and ChemCheckSlides

32. Miniaturization based on LBL technology

33. INTERPOLYELECTROLYTE COMPLEX AT WATER/SOLID INTERFACE layer-by-layer (LBL) polyelectrolyte film solid support PC A typical plot H2O H2O Thickness of LBL film, nm PA Number of layers

34. LBL 4 layers

35. DILUTED SOLUTIONS: PC conc.  1% SEMI-DILUTED SOLUTIONS: PC conc.  1%

36. LBL FILM FORMATION System with preformed NP/PC complex Simple binary system PC NP PA Complex No control of NP density within a layer Easy control of NP density within a layer

37. LATEXES BILAYER VESICLES LINEAR POLYELECTROLYTES POLYMER MICRO AND NANO GELS OBJECTS TO BE INCLUDED IN LBL FILMS METAL SOLS CARBON NANOTUBES SEMICONDUCTOR NANOPARTICLES PROTEINS INCLUDING ENZYMES AND ANTIBODIES AND OTHER CHARGED MICRO AND NANO STRUCTURES

38. _ 2e PHENOL TYROSINASE BIOSENSOR Phenol 1/2 O2 Tyrosinase A principle of analysis H2O 1/2 O2 Catechol Graphite electrode Tyrosinase H2O o-Quinone

39. DESIGN OF ELECTRODE FOR PHENOL BIOSENSOR Paste electrode LBL electrode Graphite LBL film: polyelectrolyte/enzyme Paste coat: carbon-black/ vaseline/enzyme

40. Analytical parameters for (PC/Tyr)n electrodes Electrode: (PC/Tyr)8 Linear range: 2·10-8 – 1·10-5 М Detection limit: 6nМ Sensityviyty 0,4А/(М·cm2)

41. Operation Stability for (PC/Tyr)n electrodes Electrode: (PC/Tyr)8 Вариационный коэффициент для 30 измерений за 8 часов: 6%

42. ChO biosensor

43. Electrodes design: different possibilities

44. Antibody immobilization using polyanion (PAMPS) pretreatment «Grid» ofPAMPS after washing inPBST p = 0,12 ± 0,01 V = (0,10 ± 0,01)· 106 nm3/mkm2 Antibodyadsorption on polymer «grid» p = 0,53 ± 0,15 V = (6,3 ± 1,4)· 106 nm3/mkm2

45. 2 3 4 1 Working with DNA microarrays Target preparation Probe preparation Data analysis

46. DNA array using MOLEGO approach A – Polymer B – Oligonucleotide with polymer C – Polymer with DNA (non-specific interaction) D – Polymer with DNA (specific interaction)

47. ChemCheck-1 (EasyCheck beta machine)Slides

48. Neurotoxin Analyzer – ChemCheck 1

49. Biochemical reactions Inhibition stage ChE + Inhibitor  ChE(inactive) ChE Biocatalytic stage Butyrylcholine  choline + butyryc acid ChO Biocatalytic stage Choline +2O2 + H2O  betaine + 2H2O2 Amperometric detection using a Pt-Ag/AgCl combined electrode The anodic oxidation of hydrogen peroxide at +600 mV H2O2 O2 + 2H+ + 2e- ChemCheck Scientific Basis

50. Equipment – Alpha Version