Diagnostics Alan S. Rudolph PhD MBA Chief Executive Officer Adlyfe Inc. Rockville, MD Innovation is in our blood Diagnostics

Background and Perspective Aggregate nature of misfolded prion (PrPsc) protein presents fundamental challenges in detection and sample preparation for wide variety of risk materials including blood Adlyfe has developed specific and sensitive ligands that exploit prion folding behavior for direct detection of PrP SC and are developing high throughput kits for diagnostics and screening. We have shown presymptomatic antemortem detection of PrP SC in blood and tissue using Adlyfe proprietary peptide Pronucleon ligands The lack of controlled studies and matched samples to correlate etiology of potential disease from risk materials in animals (over expected timecourse of disease) limits progress Standardized source materials for testing would accelerate development of needed tests

Diagnostics Adlyfe Pronucleon Ligands Mimic Folding and Detect PrP sc Directly PrP Adlyfe Pronucleon Ligands  -helix  -sheet PrP sc (TSE) Sequence specific peptide ligands with conformationally sensitive fluorescent labels detect PrP SC directly Folding and Aggregation of PrP SC Infectivity and Disease  -helix  -sheet

Diagnostics Adlyfe ligands associate with PrP SC and transduce signal Adlyfe ligands associate with PrP SC and transduce signal Ligands seed nucleation reaction amplifying signalLigands seed nucleation reaction amplifying signal Unparalleled Sensitivity through Novel Amplification

Diagnostics Adlyfe Ligands Enable Blood Test for BSE BSE detection in blood plasma (red) samples were also confirmed positive by western blot in matched brain tissue Negatives (blue) from blood plasma were confirmed negative by western blot of matched brain tissue). Two false positives (pink) were later confirmed positive by western

Diagnostics Detection in Different Animals Under Different Modes of Infection

Diagnostics Threshold of Detection Based upon calculations by Dr. Paul Brown of blood levels of prion in disease we estimate we are detecting in the fM range or ng PrP-Sc approaching the equivalent of one IU Concentration of prion protein (pM) Fluorescence in infected samples Advanced diagnostics Conventional diagnostics Infectivity: 1 IU = 3fM= 200,000PrP

Diagnostics TSE Test Kit 96-well polypropylene plate pre-seeded with target peptide Sequence specific ligands would create blood screen and diagnostic kits for BSE, vCJD and other neurodegerative protein misfolding diseases We are preparing to produce kits for validation and approval with USDA, FDA, EFSA, MHRA

Diagnostics Summary and Plans More sensitive detection of PrP SC will enable surveillance of risk materials and reduce risk of transmission Adlyfe ligands can detect misfolded PrP SC in risk materials including brain tissue and blood and demonstrate presymptomatic, ante mortem detection of disease We are in discussions with strategic partners to develop a high throughput diagnostic and screening test for detection and removal applications for risk materials Future R&D will further exploit detection of protein folding in neurodegenerative diseases