1. ARCHITECT cSystems & AEROSET System
QUANTIA β2-Microglobulin
6K39-40
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, version 1

2. 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
Additional Assay Parameter Configuration Tips
Questions and Answers
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

3. QUANTIA β2-Microglobulin 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, , version 1

4. Assay Disk Information
The QUANTIA assays are available for use on the AEROSET and ARCHITECT cSystems (i.e. c4000, c8000, and c16000).
There will be no AEROSET assay disk available. Assays can be manually configured.
Perform assay configuration per the parameters 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, , version 1

5. Assay Disk Information
ARCHITECT cSystems assay disks containing the QUANTIA Assays are available for easy loading of the
β2-Microglobulin parameters (B2M), Assay number 2903.
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

6. Clinical Utility of QUANTIA β2-Microglobulin
The QUANTIA β2-Microglobulin immunoassay is intended for the quantitative determination of β2-Microglobulin (B2M) 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 as an aid in the diagnosis of active rheumatoid arthritis and kidney disease.
Source(s): Quantia β2-Microglobulin 6K39 Package Insert
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

7. Clinical Utility of QUANTIA β2-Microglobulin
β2-Microglobulin is a transmembrane protein found on all cells that carry human leukocyte antigen (HLA); however, its precise immunological function is unknown.
As a result of metabolism and degradation of HLA, B2M appears in its free form and can be found at low concentrations in serum, urine, and other body.
B2M is cytotoxic to lymphocytes in the presence of complement and may be involved in the recognition phase of the immune response.
Source(s): Quantia β2-Microglobulin 6K39 Package Insert, FBR
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

8. Clinical Utility of QUANTIA β2-Microglobulin
Free B2M is a product of cell breakdown
After filtration through the glomeruli B2M is reabsorbed and catabolized by the proximal tubular cells.
Decreased glomerular filtration is associated with high serum levels of B2M, whereas tubular insufficiency is associated with normal serum and high urine levels.
Source(s): Quantia β2-Microglobulin 6K39 Package Insert, FBR
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

9. Clinical Utility of QUANTIA β2-Microglobulin
Increased Serum B2M levels are associated with:
Renal dysfunction
Lymphoproliferative disorders
Myeloma
Rheumatoid arthritis
Viral infections
Source(s): Quantia β2-Microglobulin 6K39 Package Insert, FBR
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

10. QUANTIA β2-Microglobulin Reaction Methodology
Step 1: Sample containing B2M is added to Reagent 1 (Reaction Buffer)
Step 2: Reagent 2 containing Antibody (anti-human ß2-microglobulin) coated Latex Particles is added to the sample/Reagent 1 mix
Sample/ Reagent 1
Reagent 2 ↑
B2M ↑
Latex
<200nm Diameter ↑
B2M Specific Antibody
Step 3: When sample containing B2M is added to R1 and mixed with the antibody in R2 insoluble aggregates form. The degree of agglutination is directly proportional to the concentration of B2M in the sample. This is determined by measuring the decrease of transmitted light caused by the aggregates at a wavelength of 572 nm.
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

11. Quantia β2-Microglobulin Methodology
Turbidimetric/Immunoturbidimetric
Available in the CAP Tumor Marker Survey (TM/TMX 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, , version 1

12. QUANTIA β2-Microglobulin Sample Handling
Specimen Requirements:
Serum
Ensure complete clot formation has taken place prior to centrifugation.
Separate serum from red blood cells as soon after collection as possible.
Gel separator tubes have not been tested.
There is no specification for tube type (glass or plastic)
Plasma
Acceptable anticoagulant – EDTA
Ensure centrifugation is adequate to remove platelets.
Separate plasma from red blood cells as soon after collection as possible.
Gel separator tubes 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, , version 1

13. QUANTIA β2-Microglobulin Sample Handling
Serum and Plasma
Analyze fresh specimens if possible
Sample Volume = 3.0 L
Specimen storage:
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
1 day
3 days
6 months
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

14. QUANTIA β2-Microglobulin 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 – 30 days (720 hours)
REF 6K39-40
Kit Configuration R1
2 x 11 mL R2
2 x 8 mL
110 tests/kit
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

15. QUANTIA β2-Microglobulin Calibration
Use the QUANTIA β2-Microglobulin Standard
6K47-02
Calibrator values are determined and traceable to the 1st WHO International Standard for β2-Microglobulin, National Institute for Biological Standards and Control (NIBSC) code B2M.
REF
Name
Size
β2-Microglobulin Standard Cal 1
3 x 1 mL
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

16. QUANTIA β2-Microglobulin Calibration
Do not freeze calibrators.
The single calibrator is prepared with a purified human β2-Microglobulin in a Hepes buffer solution.
The concentration of the calibrator is indicated on the QUANTIA β2-Microglobulin Standard vial label in mg/L.
Calibrator is 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, , version 1

17. QUANTIA β2-Microglobulin Calibration
The assay must be calibrated using a full calibration
(6 point calibration) procedure by using a water blank and the QUANTIA β2-Microglobulin Standard.
The cSystems and AEROSET will automatically prepare five (5) different concentration levels of calibrators from the one (1) calibrator provided.
If manually configuring the QUANTIA β2-Microglobulin assay, populate the system parameter fields as they are identified in the assay parameters packaged with the reagent.
Calibration is stable for 31 days (744 hours) and is required with each change in reagent lot number.
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

18. QUANTIA β2-Microglobulin 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, , version 1

19. QUANTIA β2-Microglobulin Calibration Curve - AEROSET
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

20. QUANTIA β2-Microglobulin Quality Control
Uses the QUANTIA PROTEINS Control, K53-21, 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)
1 x 3 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, , version 1

21. QUANTIA β2-Microglobulin Quality Control
Do not freeze controls.
The controls are prepared from human sera containing A-1-acid glycoprotein, alpha-1-antitrypsin, β2-Microglobulin, and gentamicin sulfate.
The control concentrations are indicated on the corresponding control data sheet in mg/L.
The lyophilized controls should be reconstituted with 1.0 mL of deionized 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.
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

22. QUANTIA β2-Microglobulin Performance Data
Reportable Range:
0.250 to mg/L
Limit of Quantitation (LOQ) = mg/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) = mg/L
LOD is defined as the mean concentration of an analyte-free sample + 2 SD, where SD is the within-run standard deviation.
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For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

23. QUANTIA β2-Microglobulin Product Information
*Expected Range:
0.97 – 2.64 mg/L by RIA technique
*Forman DT. Beta2-microglobulin. In: Methods in clinical chemistry. Pesce AJ, Kaplan LA. Mosby, St. Louis, USA, 1987: 68-72
Concentrations are independent of body mass and sex but are slightly increased in elderly people
Each laboratory should establish its own normal range
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

24. QUANTIA β2-Microglobulin Interference
Interference studies were conducted using an acceptance criteria of +/- 5% deviation from the target value. RF +/- 10%
Interfering Substance
Interferent Concentration
Target (mg/L)
Observed
(% of Target)
Lipemia(chyle)
2.18 AU/cm at 660 nm
2.15
99.7
Lipemia (triglyceride)
1327 mg/dL
(14.99 mmol/L)
2.24
100.45
Bilirubin
20.8 mg/dL
(355.0 µmol/L)
2.13
100.28
Hemoglobin
482 mg/dL (4.8 g/L)
2.13 mg/L
99.95
*Rheumatoid factor
240 IU/mL (240,000 IU/L)
320 IU/mL (320,000 IU/L)
4.62
4.74
109.1
112.1
*Claim of RF interference less than 10% up to 288 IU/mL extrapolated mathematically from 10 spiked interferent levels.
2 of the levels around the claim are shown here.
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For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

25. QUANTIA β2-Microglobulin Precision
Representative data from precision study are summarized
below. *Control level 3 is spiked human serum.
The precision acceptance criteria for the B2M assay is
< 6% Total CV.
Representative Data
LEVEL N
Concentration Mean
(ng/mL)
CV (%)
Within Run
Total
Quantia Proteins Control Level 1 (6K53-21) 50
1.0
2.1
2.9
Quantia Proteins Control Level 2 (6K53-21)
5.3
0.7
0.9
*Control Level 3
13.1
0.6
1.2
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

26. QUANTIA β2-Microglobulin Method Comparison
Serum results from the Quantia β2-Microglobulin assay on an ARCHITECT c System were compared with the results on the AEROSET system.
Serum results from the Quantia β2-Microglobulin assay on an AEROSET system were compared with results from ILab 900 System.
ARCHITECT cSystem vs.
AEROSET
AEROSET vs.
Comparative Method N 66
105
Y – Intercept
-0.43
0.59
Correlation Coefficient
0.9996
0.9986
Slope
0.999
0.876
Range (mg/dL)
1.34 – 71.1
1.01 – 101.1
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

27. QUANTIA β2-Microglobulin Product Summary
Assay Method: Turbidimetric /
Immunoturbidimetric
Reagent LN: K39-40
Reaction Mode: End-Up
Calibration:
Spline (Aeroset) Linear (cSystems)
QUANTIA B2M Standard
6K47-02
cSystems Reagent Volumes:
R1 Reagent: 150 L
R2 Reagent: 120 L
AEROSET Reagent Volumes:
Water: 20 L
Sample Volume: 3.0 L
Acceptable specimen: Serum/Plasma
Assay Stability:
Onboard stability = 30 days
Calibration stability = 30 days
Precision: Total CV < 6%
Reportable Range:
0.250 to mg/L
LOQ:
0.250 mg/L
LOD:
0.046 mg/L
REF
REF
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

28. QUANTIA β2-Microglobulin Additional Assay Parameter Configuration Tips
AEROSET Assay:
The AEROSET software calculates differently than the Architect cSystem software. AEROSET uses the linear high concentration considering dilutions and total reaction volumes). Because of this, mg/L is entered for C1, C2, C3, C4, and C5 calibrator concentrations.
The calibrator concentrations in the assay parameter table must be used for this assay due to the calculations used by the AEROSET software.
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

29. QUANTIA β2-Microglobulin Additional Assay Parameter Configuration Tips
cSystems Assay:
The Architect cSystems use the highest calibrator concentration (4 mg/L). This calibrator concentration should be entered for all calibrators, C1-C5.
The Architect cSystem software then calculates each calibrator concentration mathematically based upon dilutions by diluent and in reduction of sample volumes. This calculation also factors in reagent/sample volume ratios vs. total reaction volume.
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For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1

30. Not for customer distribution
Questions & Answers
Why is the Dil 2 a concentration protocol instead of a dilution?
The Dil 2 protocol uses 10x more sample (30 uL versus 3 uL) to increase the sensitivity for use with urine samples. Use of the dilution extends the LOQ to mg/L.
Not for customer distribution
For information only unless accompanied by the Lotus Notes form that contains electronic signatures. c8000 Project Notebook, , version 1