Confidential Extending the Armored RNA Technology to DNA Cindy WalkerPeach, Ph.D. Director, Development and Operations Ambion Diagnostics BSI EN ISO9001:2000 Certified BSI Registration Number: FM Quality System Regulations (21CFR§820) Medical Device Manufacturing Owner/Operator Number:

Confidential Armored RNA -> Armored RNA Quant Packaging System Armored RNA ® Armored RNA Quant ™

Confidential Armored DNA Proof of Principle – Concept Armored DNA Armored RNA Quant ™ Armored DNA

Confidential DNA Controls & Standards Commonly Used DNA plasmids Infectious or attenuated / inactivated viruses Patient samples PCR amplicons

Confidential Armored DNA Proof of Principle – DNA Packaging of two Targets Packaging of two unrelated sequences 900bp Armored DNA Armroed RNA control Ladder 239bp PCR product Armored DNA Armroed RNA control Ladder 371bp PCR product 900bp

Confidential Armored DNA Proof of Principle - Resistant to DNase 1 ug of PCR target or 5 ug of Armored DNA Internal Control was treated with and without Turbo DNase for 1 hour DNA DNA Marker Armored Armored DNA DNA No DNase +DNase

Confidential Armored DNA Proof of Principle – DNA Protection DNA protected from nuclease degradation AD (Packaged,Extracted) Extracted DNA fragment DNA fragment Ladder 239bp AD (Packaged,Extracted) Extracted DNA fragment DNA fragment Ladder 371bp

Confidential Armored DNA Proof of Principle - QC Phosphate Assay HPLC Assay DNA Integrity Functional Testing (PCR)

Confidential Armored DNA Proof of Principle – Quantification Phosphate Assay (copies/uL) qRT-PCR HBV Assay (copies/uL) HBV 8.0 x10 10 (±0.65 x ) 8.6 x % Difference

Confidential Armored DNA Heat Stability Profile AD Internal Control Control 50 C 60 C 70C ARQ Armored DNA ARQ Armored DNA

Confidential Armored DNA Detection Over a Linear Range Parallel assays of AD HBV serial curve (42 to pg) Test with two sets of primers Direct input of samples, 2 ul per reaction in duplicate

Confidential Armored DNA Initial Stability in Plasma 4 copies/uL (AD HBV 239bp) incubated in plasma (1 x 10 3 copies extracted in duplicates) Stable 7 days at 37°C Stable 14 days at 25°C Stable 1x freeze-thaw CV < 4%

Confidential Armored Technology Armored DNA summary Armored Technology compatible with DNA Packaging of 239 and 371bp double stranded DNA up to an undefined limit Stable in Plasma — 7 days at 37°C — 14 days at 25°C — 1x freeze-thaw Stable in TSMIII — 7 days at 37°C — 7 days at 25°C — 14 days at 4°C — 1x freeze-thaw

Confidential Armored DNA Ideal Controls and Standards Stable – Nuclease resistant Quantifiable – Armored DNA referenced to a NIST standard Homogeneous – single, sequence defined target Compatible – compatible with broad range of assay technologies Flexible – allows for addition to human matrix of choice Customizable – user defined sequences, genotyping Non-infectious – ease in handling and shipping