ynthetic and Systems iology orum on Microbial Threats arch Stephen Albert Johnston
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1 ynthetic and Systems iology orum on Microbial Threats arch Stephen Albert Johnston
2 HTP Screening Systems Based on Synthetic Genes and Proteins Sequence Genes Proteins Modifications Omics Integration (Systems) Manipulation Synthetic / Systems Capabilities Useful Complexity of Useful Things Complexity of Life Product
3 Proteome Screening for 1 1 Vaccines n Genes Proteins n Addressable Array Pattern
4 Screening Sera on Proteome Arrays sera All Antibody Reactive Proteins Vaccine Candidates Organism % proteins on the array % Reactive Brucella melitensis % 0.92% Burkholderia pseudomallei % 14.11% Coxiella burnetii 83.99% 0.57% Coxiella burnetii (strain:nine mile- RSA493) 75.00% 2.21% Francisella tularensis(schus4) 100% 2.24% Mycobacterium tuberculosis (H37Ev) % 0.32% Plasmodium falciparum 43.49% 9.21% Plasmodium vivax 1.65% 0.33% Pseudomonas aeruginosa 4.37% 1.00% Vaccinia(Western Reserve strain) 94.17% 26.46% Varicella zoster (Chickenpox, Shingles) 94.52% 31.51%
5 But for T-cells IVT Genes Proteins 1704 Kathryn Sykes
6 T-cell Responses to the Proteome infect 1 T-cells Kathryn Sykes/Rick Lyons
7 Expression Library Immunization Challenge with Pathogen All Genes Kathryn Sykes/Linda Dixon
8 Parapox and B2L genes Genes 250 Kathryn Sykes Michael McGuire
9 B2L
10 B2L
11 Evolutionary Pharmaco Remediation History Now Inflammatory Aspirin Premise: Genes from 5 10k years ago are not right Cholesterol Statin Sex Viagra Innate Immunity B2WL
12 Synthetic / Systems Capabilities Sequence Genes Proteins Modifications Omics Integration (Systems) Manipulate Antibodies For Human Proteome $750M + 10 years
13 Progress Thru Chemistry Antibodies Backwards Protein Splenocytes Monoclonal αβ 3 months $3K Peptide 1 Peptide 2 Linker Array of Human Proteins Protein X Is the Ligand for Protein X 5 days $1K
14 AKT1 Synbody K D = 1.5 nm Binds AKT1 in presence of 2,500 times excess protein Selectively binds AKT1 vs AKT2 vs AKT3 Chris Deihnelt
15 Antibacterial/AntiViral Therapeutic Development Bacteria Or Virus Bind to 10,000 peptide library Select Binding or Lytic Peptides High affinity Synbody Domenyuk, V., et. al, manuscript in preparation. Chris Deihnelt Zhao Zhang Valeriy Domenyuk
16 ImmunoSignaturing * Antibod y Spot Fluor Subject 1 14
17 Arrays are 1x3 glass microscope slides printed in a two-up format Reproducibility = 0.96 to R 2 Detection limit = 1.12-fold Scatterplot shows distinction between 3 naïve and 3 infected mice.
18 Normal donors Valley Fever patients Flu vaccine recipients Day 21
19 Immmunosignatures Applied to Chronic Disease Normal Glioblastoma (untreated) Unsupervised Sorting Other Diseases Analyzed Alzheimer s Disease (Published) Diabetes, T1, T2 Lupus Pancreatitis Cancers Breast Lung Ovarian Pancreatic Panins Myeloma AUC = Phil Stafford Bart Legutki
20 Chip-based High Density Diversity Neal Woodbury J. Rajasekaran
21 Current 100k Peptide Chips/cm 2 15 features 3 features 100k random sequences, 12 amino acids long, 8 different amino acids 3/17/
22 Tiling the Human Pathogen Proteome on a Single Chip 1000 human pathogens represented Tiled peptides 1000 human pathogens represented Tiled Rapid peptides identification Rapid identification Possible support for vaccine Possible support for vaccine development Testing the level of vaccine immune Testing the level of vaccine immune response response tularemia flu anthrax small pox A potentially transformative platform for global biosurveillance 3/17/
23 Uses 1M+ elements / cm HTP production 1B Low Cost All Human Proteins Tiled as Peptides 10 8 ligands/chip: αβ on Chip All Human Pathogens Tiled as Peptides
24 Future is Bright HTP Genomics Proteomics Synthetic/ Systems Knowledge Useful Things Inventive HTP Assays Chemical Diversity
25 Credits Phil Stafford Chris Diehnelt Kathryn Sykes Zhao Zhang Bart Legutki George Poste Neal Woodbury
26 Cancer Leading Killer in US $250B / Year WHO: Worldwide Epidemic Pathogen Contribution Unknown
27 Prophylactic Cancer Vaccine to Prevent All Cancers
28 Tumor Sequencing Tumor # Normal DNA RNA Breast 17 Value Diagnostic Geevac 2 Knowledge
29 Complexity
30 Personal Therapeutic Vaccine Personal Vaccine HTP Sequencing of Tumor 1 week Neotumor Antigens Create Vaccine 2 days Requirements: Early Detection Sequence from Small Sample Time: short Expense: cheap
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