1. Identify Patient Zero of a Zombie Apocalypse With the Power of an ELISA!

2. Bio-Rad Curriculum and Training Specialist
Tamica Stubbs
Damon Tighe
Sherri Andrews, Ph.D.
Leigh Brown

3. TV or real life?
The Walking Dead show
Feb 11,2014 Atlanta

4. Zombiefication Method of infection Symptoms Treatment
Bodily fluid contact
Airborne
Foodborne
Symptoms
Undead
Hungry for flesh
Treatment
None known

5. Introduction to Immunology, Antibodies and ELISA
Fluid swapping and disease spreading!
Indirect ELISA to narrow down who were the initial carriers of the disease

6. Crash Course in Immunology and ELISA
Immune Response
C. Macrophage
A. Pathogen
Zombie virus
D. Macrophage
B. B cells
F. T cell
E. Macrophage
G. B cell
J. Antibodies attach to pathogen
Antibodies are ineffective
Against Zombie Virus
H. Memory B cells
I. Plasma cells

7. Crash Course in Immunology and ELISA
Antibody Structure
ELISA tests are
based on immune
system antibody
molecules
Light chain
Heavy chain
Disulfide bonds
Antigens

8. ELISA Enzyme-Linked Immunosorbant Assay (indirect)
Add the purified antigen to all the wells. Incubate for 5 min. Rinse
Add serum antibodies (student samples) to the appropriate wells. Incubate for 5 min. Rinse
Add the enzyme-linked antibody to all wells. Incubate for 5 min. Rinse
Add enzyme substrate to all wells. Incubate for 5 min.

9. Swapping Fluids and Disease
Swap with 1 person at your table (record who)
Add your 750 µl together, mix and each take 750 µl back

10. Swapping Fluids and Disease
Swap with 1 person at the closest table (record who)
Add your 750 µl together, mix and each take 750 µl back

11. Swapping Fluids and Disease
Swap with 1 person at the another table (record who)
Add your 750 µl together, mix and each take 750 µl back

12. Testing for who is infected
ELISA Time!
Testing for who is infected

13. ELISA Time! Testing for who is infected
Your ID #
Label Tubes

14. ELISA Time! Testing for who is infected
Your ID #
Add 50 µl “+” (Antigen)

15. Add 50ul “-” (wash buffer)
Your ID #
Your ID #
Your ID #

16. Add 50ul Bodily Fluid
Your ID #
Your ID #
Your ID #

17. ELISA Time! Testing for who is infected
Your ID #
Incubate: 5 minutes

18. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

19. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

20. ELISA Time! Testing for who is infected
Polystyrene (plate)
Protein (Antigen)
Concept Corner
Hydrophobic
interactions

21. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

22. ELISA Time! Testing for who is infected
Your ID #
Wash with Buffer (contains Tween20)

23. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

24. ELISA Time! Testing for who is infected
Your ID #
Wash with Buffer (contains Tween20)

25. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

26. ELISA Time! Testing for who is infected
Add 50 µl “PA” (Primary Antibody)
Your ID #
Your ID #

27. ELISA Time! Testing for who is infected
Incubate: 5 minutes
Your ID #
Your ID #

28. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

29. ELISA Time! Testing for who is infected
Concept Corner
Your ID #
Your ID #

30. ELISA Time! Testing for who is infected
Variable region
Light chain
Heavy chain
constant region
3,368,600 antibody combos
Concept Corner

31. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

32. ELISA Time! Testing for who is infected
Wash with Buffer
Your ID #
Your ID #

33. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

34. ELISA Time! Testing for who is infected
Wash with Buffer
Your ID #
Your ID #

35. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

36. ELISA Time! Testing for who is infected
Add 50ul “SA” (Secondary Antibody)
Your ID #
Your ID #

37. ELISA Time! Testing for who is infected
Your ID #
Incubate: 5 minutes

38. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

39. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

40. ELISA Time! Testing for who is infected
Your ID #
Amplification of signal
Introduce HRP (Horseradish Peroxidase) for colormetric detection
Concept Corner

41. ELISA Time! Testing for who is infected
Concept Corner
Antigen = chicken gamma globulin
Primary Antibody = anti-chicken polyclonal antibody
Secondary Antibody = goat anti-rabbit conjugate to HRP

42. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

43. ELISA Time! Testing for who is infected
Wash with Buffer
Your ID #
Your ID #

44. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

45. ELISA Time! Testing for who is infected
Wash with Buffer
Your ID #
Your ID #

46. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

47. ELISA Time! Testing for who is infected
Wash with Buffer
Your ID #
Your ID #

48. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

49. ELISA Time! Testing for who is infected
Add 50 µl “Sub” (HRP substrate)
Your ID #
Your ID #

50. ELISA Time! Testing for who is infected
Incubate: 5 minutes
Watch for color change!
Your ID #
Your ID #

51. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

52. ELISA Time! Testing for who is infected
Concept Corner
Your ID #

53. ELISA Time! Testing for who is infected
Concept Corner

54. Deducing who was originally infected

55. Deducing who was originally infected
Everyone Stand UP! If your ELISA was negative sit down

56. Deducing who was originally infected
Round 1
If someone you shared with just sat down, also sit down
Round 2
Round 3

57. Deducing who was originally infected
Who is left?

58. Bio-Rad’s Immuno Explorer Lab
What did we use today?
Bio-Rad’s Immuno Explorer Lab
EDU

59. Other Zombie themed learning ->
Texas instruments has a program for modeling (via graphing) the spread of a Zombie Virus:
Bio-Rad’s Secrets of the Rainforest Kit can be used in a Zombie apocalypse story where students use column chromatography to isolate the Zombie Green Fluorescent Protein.