Biotechnology Explorer™ | explorer.bio-rad.com 1 Identify Patient Zero of a Zombie Apocalypse With the Power of an ELISA!

Biotechnology Explorer™ | explorer.bio-rad.com 2 Instructors - Bio-Rad Curriculum and Training Specialists  Sherri Andrews, Ph.D.  Damon Tighe  Leigh Brown, M.A.

Biotechnology Explorer™ | explorer.bio-rad.com 3 TV or real life? Feb 11, 2014 Atlanta The Walking Dead show

Biotechnology Explorer™ | explorer.bio-rad.com 4 Zombiefication  Method of infection –Bodily fluid contact –Airborne –Foodborne –Other?  Symptoms –Undead –Hungry for flesh  Treatment –Under investigation

Biotechnology Explorer™ | explorer.bio-rad.com 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

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

Biotechnology Explorer™ | explorer.bio-rad.com 7 Light chain Heavy chain Disulfide bonds Antigens Antibody Structure ELISA tests are based on immune system antibody molecules Crash Course in Immunology and ELISA

Biotechnology Explorer™ | explorer.bio-rad.com 8 ELISA E nzyme- L inked I mmuno s orbant A ssay (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.

Biotechnology Explorer™ | explorer.bio-rad.com 9 Time to Swap Fluids and disease!

Biotechnology Explorer™ | explorer.bio-rad.com 10 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

Biotechnology Explorer™ | explorer.bio-rad.com 11 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

Biotechnology Explorer™ | explorer.bio-rad.com 12 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

Biotechnology Explorer™ | explorer.bio-rad.com 13 ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 14 Your ID # Label Tubes ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 15 Your ID # Incubate: 5 minutes ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 16 Tap out liquid Your ID # ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 17 Your ID # Wash with Buffer (contains Tween20) ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 18 ELISA Time! Testing for who is infected Tap out liquid Your ID # Repeat

Biotechnology Explorer™ | explorer.bio-rad.com 19 Your ID # Add 50 µl + Control Add 50ul – Control Add 50ul of Bodily Fluid ELISA Time! Testing for who is infected Your ID #

Biotechnology Explorer™ | explorer.bio-rad.com 20 Your ID # Incubate: 5 minutes ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 21 Your ID # Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 22 Your ID # Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 23 RR HH Polystyrene (plate) Protein (Antigen) Hydrophobic interactions Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 24 Tap out liquid Your ID # ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 25 Your ID # Wash with Buffer (contains Tween20) ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 26 Tap out liquid Your ID # ELISA Time! Testing for who is infected Repeat

Biotechnology Explorer™ | explorer.bio-rad.com 27 Your ID # Add 50 µl “PA” (Primary Antibody) ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 28 Your ID # Incubate: 5 minutes ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 29 Your ID # Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 30 Your ID # Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 31 Variable region Light chain Heavy chain constant region 3,368,600 antibody combos Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 32 Concept Corner Antigen = chicken gamma globulin Primary Antibody = anti-chicken polyclonal antibody Secondary Antibody = goat anti-rabbit conjugate to HRP ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 33 Tap out liquid Your ID # ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 34 Your ID # Wash with Buffer ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 35 Tap out liquid Your ID # ELISA Time! Testing for who is infected Repeat

Biotechnology Explorer™ | explorer.bio-rad.com 36 Your ID # Add 50ul “SA” (Secondary Antibody) ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 37 Your ID # Incubate: 5 minutes ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 38 Your ID # Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 39 Your ID # Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 40 Your ID # Amplification of signal Introduce HRP (Horseradish Peroxidase) for colormetric detection Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 41 Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 42 Tap out liquid Your ID # ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 43 Your ID # Wash with Buffer ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 44 Tap out liquid Your ID # ELISA Time! Testing for who is infected Repeat x2

Biotechnology Explorer™ | explorer.bio-rad.com 45 Your ID # Add 50 µl “Sub” (HRP substrate) ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 46 Your ID # Incubate: 5 minutes Watch for color change! ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 47 Your ID # Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 48 Your ID # Concept Corner ELISA Time! Testing for who is infected

Biotechnology Explorer™ | explorer.bio-rad.com 49 Deducing who was originally infected

Biotechnology Explorer™ | explorer.bio-rad.com 50 Deducing who was originally infected Everyone Stand UP! If your ELISA was negative sit down

Biotechnology Explorer™ | explorer.bio-rad.com 51 Deducing who was originally infected  Round 1 –If someone you shared with just sat down, also sit down  Round 2 –If someone you shared with just sat down, also sit down  Round 3 –If someone you shared with just sat down, also sit down

Biotechnology Explorer™ | explorer.bio-rad.com 52 Who is left? Deducing who was originally infected

Biotechnology Explorer™ | explorer.bio-rad.com 53  Jeff Lukens Guest Zombie Instructor

Biotechnology Explorer™ | explorer.bio-rad.com 54 Guest Zombie Instructor  Jeff Lukens

Biotechnology Explorer™ | explorer.bio-rad.com 55 Using Zombies to Model the Spread of an Infection!  Jeff – a few slides of TI stuff can start here

Biotechnology Explorer™ | explorer.bio-rad.com 56 The Zombie Autopsies by Dr. Steven Schlozman, MD

Biotechnology Explorer™ | explorer.bio-rad.com 57 The Scenario  A new virus has been discovered that affects the brains of humans, turning them into zombies! Epidemiologists are trying to figure out what to do to help stop the outbreak. The virus is airborne, being transported in saliva droplets of affected individuals. The zombies are attacking healthy humans as their primary food source and new zombies are showing up at an alarming rate.

Biotechnology Explorer™ | explorer.bio-rad.com 58 Connections to Solid Science Practices  Asking questions and defining problems –What happens to the brain to cause “zombification”? –How does a disease spread through a population?  Using mathematical and computational thinking –Graphically and numerically evaluate the spread of a disease in a population  Developing and using models –Brain models –Graphical representations of data  Analyzing and interpreting data –Using mathematics and graphical literacy to evaluate dat

Biotechnology Explorer™ | explorer.bio-rad.com 59 Symptoms of the Infection  Affected Cerebellum: Zombies clumsily shuffle forward.  Affected Hypothalamus: Zombies have insatiable appetites.  Affected Frontal Lobe: Zombies are poor problem solvers.  Affected Amygdala: Zombies aren't nice. They are full of rage.

Biotechnology Explorer™ | explorer.bio-rad.com 60 Normal Dude—Pre-Infection

Biotechnology Explorer™ | explorer.bio-rad.com 61 Infected Dude—Early Infection

Biotechnology Explorer™ | explorer.bio-rad.com 62 Sick Looking Dude—Stage 2 of Infection

Biotechnology Explorer™ | explorer.bio-rad.com 63 Creepy Looking Dude—Stage 3 of Infection

Biotechnology Explorer™ | explorer.bio-rad.com 64 Trying-to-Eat-You Dude—Full-Blown Infection!!

Biotechnology Explorer™ | explorer.bio-rad.com 65 Graphing the Infection Spread

Biotechnology Explorer™ | explorer.bio-rad.com 66 Zooming In For a Better Look

Biotechnology Explorer™ | explorer.bio-rad.com 67 Fill the Blocks With Zombies!!

Biotechnology Explorer™ | explorer.bio-rad.com 68 Stocking Up For the “Z” Apocalypse!

Biotechnology Explorer™ | explorer.bio-rad.com 69 Hmmm….Why are the Graphs Curvy???