AOS-HCI Comparing the effects of alligator and horseshoe crab blood on cancer cell viability

1. AOS-HCIComparing the effects of alligator and horseshoe crab blood on cancer cell viability Estelle Gong AnuVenkatachalam Ryan Wong Joshua Foo

2. Background AOS HCI Robust innate immune system of the Singapore horseshoe crab (Carcinoscorpiusrotundicauda) Infection studies on them have demonstrated that a large amount of bacteria was rapidly suppressed. Proteins found in the blood of horseshoe crabs potentially provides a more effective way of treating cancer • American alligators (A. missippiensis) have remarkable immune systems • Blood serum components • Muscle tissue peptides • Cannot develop tumors; cannot develop cancers

3. Purpose AOS HCI Elucidate specific proteins in Horseshoe crab blood that recognize and bind surface antigens/ PAMPs of cancer cells To propose potential peptide-based drugs for cancer detection & treatment • To find what components of alligator serum, if any, have anticancer properties • Testing serum components against cancer cell membrane proteins • Isolating specific

4. Hypothesis & IV/DV AOS HCI Proteins present in horseshoe crab blood recognize and bind PAMPs of cancer cells. IV: Horseshoe crab blood DV: Proteins that bind PAMPs of cancer cells • If alligator serum has anticancer properties, then proteins in alligator serum will bind to cancer cell membrane proteins. • IV: Alligator blood serum • DV: Proteins that bind to cancer cell membrane proteins

5. Experimental Design • AOS: • Test alligator blood serum on whole colorectal cancer cell cultures to confirm its effectiveness, if any, against cell viability. • Both AOS and HCI: • Separate serum/hemolymph from whole alligator and horseshoe crab blood, respectively • AOS – culture and lyse colorectal cells; HCI buys colorectal lysates • Isolate the proteins in the cell membrane using the QIAGEN protein kit. • Load affinity columns with membrane proteins and run serum/hemolymph through so proteins present in serum bind to proteins in the membrane. • Separate proteins of the cell membrane on the beads from the active components of the serum/hemolymph. • Use gel electrophoresis to categorize the active components of the whole serum/hemolymphto determine its components and makeup. • Analyse the isolated active components by MALDI-TOF or other methods.

6. Timeline • Present – February – AOS works, HCI break • AOS tests alligator blood serum on whole colorectal cancer cells • February – May –AOS & HCI works • Both test blood serum/hemolymph against colorectal cell membrane proteins • Isolate most effective components of • May – August – AOS break, HCI works

7. References