# Presentation: 818820

## Innovation is in our blood

**Chief Executive Officer**
**Adlyfe Inc.**
**Rockville, MD**
**arudolph@adlyfe.com**
**301-424-8344**

*Innovation is in our blood*

## 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 PrPSC and are developing high throughput kits for diagnostics and screening.
- We have shown presymptomatic antemortem detection of PrPSC 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

## Adlyfe Pronucleon Ligands MimicFolding and Detect PrPsc Directly 

- Adlyfe Pronucleon Ligands MimicFolding and Detect PrPsc 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**_

**Notes:**
The peptide represents the minimum plausible unit.  The minimum goal working with the first peptides is to demonstrate that conformer switching is kinetically distinguishable for spontaneous convertion vs induced.  The purpose of the winged peptides and dendrimers in the first place is to reduce spontaneous conversion to negligible amounts while retaining significant (measureable) induced conversion.  

Obviously any system must be its very nature, carefully balance the two tendencies.  Practically any functioning engineering system is in only quasi-stable equilibrium.  That’s why it works.  We’re talking both about a feedback system and an optimized one.  As long as we can distinguish (spontaneous from induced) mechanism, we can favor one or the other.

Dendrimers are a means of increasing local concentrations of the target and thus facilitating he kinetics of the transition

## Unparalleled Sensitivity through Novel Amplification

-  Adlyfe ligands associate with PrPSC  and transduce signal
- Ligands seed nucleation reaction amplifying signal

- Unparalleled Sensitivity through Novel Amplification

**Notes:**
After two decades of intense debate, it is now widely accepted that the misfolded prion protein is the essential, if not the only, component of the TSE infectious particle 

The development of a blood test for TSEs is one of the major challenges in the prion research fields.
In the U.S. alone, it is estimated that 4.5-5.3% of blood donors are deferred due to the potential for vCJD transmission.

Since vCJD is nonexistent in the US, if a blood test existed, the donor loss could be avoided.  A blood assay could also play a role in both monitoring the disease’s progression and in developing therapeutic approaches.  

.  There are several reasons why a blood based test is not yet available:
The misfolded prion protein does not elicit an immune response, preventing the development of a serological antibody test;
The absence of a nucleic acid component, eliminates the PCR approach
The compartmentalization of PrP-Sc into the CNS leads to a very low titer in blood, well below the detection limit of a conventional immunoassay.
The task is further complicated by the existence of large amts of the normal cellular for of the prion protein (PrP-C) in the blood which interferes with its detection

## 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 **

## Detection in Different Animals Under Different Modes of Infection

## 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 0.1-1 ng PrP-Sc approaching the equivalent of one IU

##  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**

## Summary and Plans

- More sensitive detection of PrPSC will enable surveillance of risk materials and reduce risk of transmission
- Adlyfe ligands can detect misfolded PrPSC 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