ARCHITECT cSystems & AEROSET System QUANTIA Alpha-1-Antitrypsin 6K99-40 US Only Launch: June 2010 REF Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29 version 1

Not for customer distribution Agenda Launch Strategy Assay Disk Information Clinical Utility Reaction Methodology Sample Handling Reagent Handling Calibration Quality Control Performance Data Interference Precision Method Comparison Product Summary Questions and Answers Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A-1-AT Launch Strategy The QUANTIA assays are available for use on the AEROSET and ARCHITECT cSystems. Assay will be available in the US only. Latin America and Canada are considered ex-US for this product. There are no differences in the application or formulation between the U.S. and non-US products. Complaint handling: Abbott has responsibility for initial complaint registration. BioKit (the vendor) will investigate Level II complaints. Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

Assay Disk Information Alpha-1-Antitrypsin parameters (A1AT, Assay number 2901) are available on the following recently released ARCHITECT cSystems assay disks. Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

Assay Disk Information There will be no AEROSET assay disk available. Assay configuration is provided in the assay specific application sheets. Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

Not for customer distribution Clinical Utility The QUANTIA Alpha-1-Antitrypsin (A1-AT) immunoassay is intended for the quantitative determination of A1-AT in human serum or plasma on the ARCHITECT cSystems (i.e. c4000, c8000, and c16000) and the AEROSET System. The assay is intended for use in clinical laboratories and is used as an aid in the diagnosis of juvenile and adult cirrhosis of the liver and pulmonary emphysema. A1-AT is a protein synthesized in the liver that inhibits several proteolytic enzymes such as elastase, chymotrypsin, trypsin and thrombin. Source(s): Quantia A1-AT 6K99 Package Insert Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

Not for customer distribution Clinical Utility A1-AT protein deficiency is a genetic predisposition characterized by varying levels of severity. When A1-AT is deficient in adults, uninhibited activity of these enzymes results in pulmonary emphysema and hepatic diseases. Among children, severe deficiency most frequently presents as liver disease, cholestasis and cirrhosis. A1-AT deficiency can be treated but not cured. Source(s): Quantia A1-AT 6K99 Package Insert, Medline Plus Online Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

Not for customer distribution Clinical Utility Low A1-AT levels can be associated with: Liver Cirrhosis Liver Tumors Inherited deficiency Nephrotic syndrome Malnutrition Other hepatic and pulmonary problems Source(s): Quantia A1-AT 6K99 Package Insert, Medline Plus Online Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

Not for customer distribution Clinical Utility Increased levels of A1-AT are seen with: Inflammation Necrosis Acute hepatitis Malignant tumors Rheumatoid disease Pregnancy Estrogen or androgen therapy Source(s): Quantia A1-AT 6K99 Package Insert, FBR, and Tietz textbook of Clinical Chemistry and Molecular Diagnostics Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Reaction Methodology Step 1: Sample containing A1-AT is added to Reagent 1 (Reaction Buffer and polyethylene glycol (PEG)). Step 2: Reagent 2 containing Antibody (goat serum anti-human Alpha-1-Antitrypsin) is added to the sample/Reagent 1 mix Sample/ Reagent 1 Reagent 2 Y Y Y Y Y + Y Y ↑ A-1-AT Specific Antibody ↑ A-1-AT Step 3: When sample containing A1-AT, diluted in the reaction buffer, is mixed with the antibody in reagent 2, insoluble aggregates form. The degree of agglutination is directly proportional to the concentration of A1-AT in the sample. This is determined by measuring the decrease of transmitted light caused by the aggregates at a wavelength of 604 nm. Y Y Y Y Y Y Y Y Y Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

Quantia A-1-AT Methodology Turbidimetric/Immunoturbidimetric Available in the CAP Diagnostic Immunology Survey (IG/IGX series) Contact CAP for the current coding. Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Sample Handling Specimen Requirements: Serum Gel separator tubes have not been tested. There is no specification for tube type (glass or plastic) Plasma Acceptable anticoagulant – EDTA and Lithium Heparin Gel separator tubes other than lithium heparin have not been tested. Note: Use fresh sample collected by standard venipuncture techniques Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Sample Handling Serum or Plasma only Analyze fresh specimens if possible Sample Volume = 2.0 L Specimen storage: *Guder WG, da Fonseca-Wollheim F, Heil W, et al. The Quality of Diagnostic Samples. Darmstadt, Germany: GIT Verlag; 2001:13-14. Repeated freeze/thaw cycles should be avoided to minimize potential protein degradation. Stored specimens must be inspected for particulates. If present, mix and centrifuge to remove particulates prior to testing. Temperature Maximum Storage Serum/Plasma *20 to 25°C 2 to 8°C -20°C 3 months 5 months Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Reagent Handling Reagent Kit Configuration Estimated calculation is based on the minimum reagent fill volume per kit. Reagent is liquid, ready-to-use, 2 reagent kit Storage 2 to 8°C. Unopened kit stable to expiration date Remove any air bubbles, if present, with an applicator stick Reagent onboard stability – 35 days (840 hours) REF 6K99-40 Kit Configuration R1 2 x 18 mL R2 2 x 5 mL 146 tests/kit Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Calibration Calibration is stable for 30 days (720 hours) and is required with each change in reagent lot number. Use the QUANTIA PROTEINS Calibrator 6K45-02 Calibration: The assay must be calibrated using a full calibration (6 point calibration) procedure by using a water blank and the five levels of QUANTIA PROTEINS Calibrator. REF Name Size Protein Calibrator 1 1 x 1 mL Protein Calibrator 2 Protein Calibrator 3 Protein Calibrator 4 Protein Calibrator 5 Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Calibration Do not freeze calibrators. The calibrators are prepared with purified alpha-1-antitrypsin. Calibrator values are determined and traceable to the International Reference Material CRM470. The concentration of the calibrators are indicated on the QUANTIA PROTEINS Calibrator value sheet. Calibrators are liquid ready to use and stable until the expiration date indicated on the package when stored at 2-8 °C. Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Calibration Curve – ARCHITECT cSystems Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Calibration Curve - AEROSET Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Quality Control Uses the QUANTIA PROTEINS Control, 6K53-21, available for use for both the ARCHITECT and AEROSET customers. QUANTIA PROTEINS Control, List No. 6K53-21 contains: This is the only control approved for use with this assay REF Control Product Name Configuration Control I Low Level (lyophilized) 3 x 1 mL Control II High Level (lyophilized) Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Quality Control The multi-constituent controls are prepared from human sera containing A-1-acid glycoprotein, alpha-1-antitrypsin, β2-Microglobulin, and gentamicin sulfate. The concentrations are indicated on the corresponding QUANTIA PROTEINS Control value sheet. The lyophilized controls should be reconstituted with 1 mL of Type II water or equivalent. The lyophilized controls are stable until the expiration date on the label if stored at 2-8 °C. Once reconstituted, the controls can be used for 15 days if stored between 2-8 °C. Do not freeze controls. Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Performance Data Serum and Plasma Expected Range: 84 to 200 mg/dL (0.84 to 2.00 g/L) Reportable Range: 25.0 to 300.0 mg/dL (0.25 to 3.00 g/L) 25.0 to 1500.0 mg/dL (0.25 to 15.00 g/L) using the on-board dilution protocol Limit of Quantitation (LOQ) = 25.0 mg/dL (0.25 g/L) LOQ represents the lowest measurable concentration at which the within-run CV < 20% and the recovery is within +/- 20% of expected value. Limit of Detection (LOD) = 0.70 mg/dL (0.007 g/L) LOD is defined as the mean concentration of an analyte-free sample + 2SD, where SD is the within-run standard deviation. Note: SI units are g/L To convert results from mg/dL to g/L, multiply mg/dL by 0.01. Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Interference Interference studies were conducted using an acceptance criteria of +/- 5% deviation from the target value except for Rheumatoid Factor which uses +/- 10%: Interfering Substance Interferent Concentration Target (mg/dL) Observed (% of Target) Lipemia (Chyle)* 2.3 AU/cm at 660 nm 145.4 99.5 Lipemia (Triglycerides) 1327 mg/dL (14.99 mmol/L) 138.5 99.8 Bilirubin 19.6 mg/dL (335µmol/L) 145.0 100.0 Hemoglobin 460 mg/dL (4.6 g/L) 144.8 99.6 **Rheumatoid factor 800 IU/mL (800,000 U/L) 137.2 107.7 *Solution manufactured to mimic chyle, which is a milky fluid consisting of lymph and emulsified fat extracted from chyme by the lacteals during digestion. **Claim of RF interference less than 10% up to 138 IU/mL interpolated mathematically from 10 spiked interferent levels. 2 of the levels around the claim are shown here. Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Precision Representative data from precision study are summarized below. See Q&A slide 28 for control levels used. The precision acceptance criteria for the A1-AT assay is < 6% Total CV. Representative Data LEVEL N Concentration Mean (mg/dL) CV (%) Within Run Total Control Level 1 50 78.0 3.4 Control Level 2 157.5 0.8 1.0 Control Level 3 231.0 0.7 Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Method Comparison Serum results from the A1-AT assay on an ARCHITECT c System were compared with the results on the AEROSET system. Serum results from the A1-AT assay on an AEROSET system were compared with results on Dade-Behring BN II ARCHITECT cSystem vs. AEROSET AEROSET vs. Comparative Method N 79 111 Y – Intercept -5.5 12.2 Correlation Coefficient 0.995 0.989 Slope 1.027 1.002 Range (mg/dL) 41.6 – 316.3 42.0 – 442.5 Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Product Summary Assay Method: Turbidimetric / Immunoturbidimetric Reagent LN: 6K99-40 Reaction Mode: End-Up Calibration: Linear QUANTIA PROTEINS Calibrator 6K45-02 cSystems Reagent Volumes: R1 Reagent: 190 L R2 Reagent: 50 L Water: 45 L AEROSET Reagent Volumes: Water: 20 L Sample Volume: 2.0 L Acceptable specimen: Serum, plasma Assay Stability: Onboard stability = 35 days Calibration stability = 30 days Precision: Total CV < 6% Reportable Range: 25.0 to 300.0 mg/dL (Conv) 0.25 to 3.00 g/L (SI) LOQ: 25.0 mg/dL (Conv) 0.25 g/L (SI) LOD: 0.70 mg/dL (Conv) 0.007 g/L (SI) REF REF Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Product Summary SmartWash: Assay parameters for ARCHITECT cSystems Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

QUANTIA A1-AT Product Summary SmartWash: Assay parameters for AEROSET System Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1

Not for customer distribution Q&A There are only 2 levels of Quantia controls, what sample was used for the 3rd level in precision testing? A 1:1 mixture of Level 1 and Level 2 was used to create a third level for this testing. END OF DOCUMENT Not for customer distribution For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, 3.6.29, version 1