Live (virulent) virus Disease Immune response What happens when you get infected? DeathSurvive (Immune) OR

Live (virulent) virus Killed or avirulent virus Disease Immune response No Disease Immune response Vaccines help you fight infections

Components of an Immune Response Antibodies Killer white cells (CTLs)

Immune surveillance The Immune Response Protects Against Cancer

Antibodies are a Key Part of our Immune System

Antibodies are important in malaria

The malaria parasite gets into our red blood cells and multiplies

The malaria parasite invades red blood cells

The malaria parasite surface is a target for antibodies

Antibodies bind to proteins on the malaria parasite surface and prevent invasion of RBC Human Antibody Malaria protein

Antibodies are important in cancer

Antibodies are a major class of drugs :

1. Orthoclone: mouse antibody (Ortho Biotech) Transplant anti-rejection 2. ReoPro: chimeric antibody (Centocor)Prevents blood clotting 3. Rituxan: chimeric antibody (Genetech) Cancer therapy/ anti arthritis $1.9billion 4. Zenapax: humanized antibody (Hoffman-La Roche) Transplant immune suppression 5. Simulect: chimeric antibody (Novartis) Transplant anti-rejection 6. Synagis: humanized antibody (Medimmune) Anti-respiratory syncytial virus 7. Remicade: chimeric antibody (Centocor) Anti-arthritis treatment 8. Herceptin: humanized antibody (Genetech) Cancer therapy $1.45billion 9. Mylotarg: humanized antibody and calechiamycin (Wyeth) Cancer therapy 10. Campath-1H: humanized antibody (Genzyme) Cancer therapy 11. Humira: human antibody (Abbott/CAT) Anti-arthritis treatment $883million 12. Xolair: humanized antibody (Genetech)Treatment of allergic asthma 13. Raptiva: humanized antibody (Genetech)Treatment of psoriasis 14. Erbitux: chimeric antibody (Imclone systems)Cancer therapy 15. Avastin: humanized antibody (Genetech)Cancer therapy $1.3billion 16. Tysabri: humanized antibody (Biogen Idec) Multiple sclerosis treatment 17. Lucentis: humanized antibody (Genetech)Macular degeneration 18. Vectibix: human antibody (Amgen)Cancer therapy 19. Zevalin: mouse antibody and Yttrium 90 (Biogen Idec) Cancer therapy 20. Bexxar: mouse antibody and Iodine 131 (Corixa) Cancer therapy FDA approved antibodies:

Antibodies’ long revenue stream spurs pharma acquisitions (Nature Biotechnolgy, Volume 11, November, 2007) Biotech Pharma Date Amount Domantis (UK) GlaxoSmithKine (UK) Jan 8, 2007 $454million Therapeutic Human Roche (Switzerland) April 2, 2007 $56.5 million Polyclonals (USA) Abmaxis (USA) Merck (USA) May 9, 2007 $80 million MedImmune (USA) AstraZeneca (UK) June 19, 2007 $15.6 billion Haptogen (UK) Wyeth (USA) Oct 5, 2007 not disclosed

Cost of getting a drug to market Clinical trials can cost up to $1 billion. The cost of the research to discover a new drug, by contrast is less than 10% of this. New drugs need Intellectual Property (IP) protection eg patents. Big Pharma is mainly interested in huge markets eg obesity, depression, dementia, cancer, autoimmune disease. Amgen’s Magic Bullet, Retuximab against blood cancer, earns them more than $3 billion per year. Amgen’s Erythropoietin (EPO), which stimulates the production of red blood cells, raises $4 billion per year. This is the drug cyclists and long distance runners and swimmers use (illegally)!

Shark Antibodies

Shark antibodies are 1/10 the size of human antibodies Shark Mab Why Shark Antibodies?

Tailored half life In humans Low cost of manufacture in microbes Extreme stability Address targets inaccessible to conventional antibodies Why Shark Antibodies?

IgG Shark Antibodies VHVH Greenberg, Flajnik et al. Nature 374 (1995) What if you could capture these shark antibodies in a test tube ? IgNAR

Generation of shark antibody library BLOOD SAMPLE TAKEN FROM SHARK LYMPHOCYTE FRACTION ISOLATED mRNA PREPARATION MADE FROM LYMPHOCYTES WOBBEGONG SHARK cDNA COPIES CORRESPONDING TO mRNA FROM ANTIBODY PRODUCING CELLS Sfi1 Not1 CDR1CDR3 Degenerate primer for mutagenesis Making a shark antibody library Could you use this big library to select antibodies for therapy ?

Selecting shark antibodies against malaria Shark library

Shark antibodies bind to key sites on malaria Shark antibody Malaria protein

Shark antibodies can block key sites on malaria

OR can we make antibodies bind better ?

Better shark antibodies block malaria invasion

Fibrosis is a major disease

CXCR4 has been associated with more than 23 types of cancers HIV-1 uses CXCR4 as a co-receptor for viral entry into host cells Critical player in development of Fibrosis Important in maintaining stem cells in bone marrow

Selecting shark antibodies against CXCR4 Shark library CXCR4 Shark antibody CXCR4 Shark antibody complex

Binding to Cancer cells 21H5 114 Block HIV infection anti-CXCR4 i-bodies do many things Bind to rat fibrocytes DAPI CD45 i-body Merge

Lekocytes (white blood cells) can cause fibrosis Fibrosis

CXCR4 controls migration of white blood cells into tissues

Where to next?

Sharks are nice

But we would rather not work with them