A1 9D92-20 304511/R5

APOLIPOPROTEIN A1

This package insert contains information to run the Apolipoprotein A1 assay on the ARCHITECT c Systems and the AEROSET System.

NOTE: Changes Highlighted

NOTE: This package insert must be read carefully prior to product use. Package insert instructions must be followed accordingly. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this package insert.

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Reagent 1

Reagent 2

December 2009 ©2004, 2009 Abbott Laboratories

1 NAME REAGENT HANDLING AND STORAGE APOLIPOPROTEIN A1 Reagent Handling Remove air bubbles, if present in the reagent cartridge, with a new INTENDED USE applicator stick. Alternatively, allow the reagent to sit at the appropriate The Apolipoprotein A1 (ApoA) assay is used for the quantitation of storage temperature to allow the bubbles to dissipate. To minimize apolipoprotein A-I (ApoA-I) in human serum or plasma. volume depletion, do not use a transfer pipette to remove the bubbles. CAUTION: Reagent bubbles may interfere with proper detection of SUMMARY AND EXPLANATION OF TEST reagent level in the cartridge, causing insufficient reagent aspiration that are synthesized in the intestine or liver need to be which could impact results. transported to tissues and organs for their varied metabolic functions. Given the hydrophobic nature of the neutral , , Reagent Storage and esters, transport and delivery via plasma Unopened reagents are stable until the expiration date when stored would not be possible without some form of hydrophilic adaptation. at 2 to 8°C. Lipids are transported by means of a series of micellar structures Reagent onboard stability is approximately 57 days if quality control known as that consist of an outer monolayer of results meet acceptance criteria. If quality control results do not meet (apolipoprotein), polar lipids, and an inner core of neutral lipids. acceptance criteria, refer to the QUALITY CONTROL section of this ApoA-I is synthesized in both the liver and intestine, while ApoA-II package insert. synthesis appears to occur only in the liver. ApoA-I and ApoA-II are the major found in HDL, with small amounts found in WARNINGS AND PRECAUTIONS . Approximately 50% of HDL mass is protein, with ApoA-I and ApoA-II constituting approximately 90% of the fraction. Precautions for Users The ratio of A-I to A-II is roughly 3:1. On average, ApoA-I levels are 1. For in vitro diagnostic use. higher in women than in men, whereas ApoA-II levels are similar. 2. Do not use components beyond the expiration date. Lipoprotein particles that contain only ApoA-I appear to stimulate 3. Do not mix materials from different kit lot numbers. cholesterol efflux (movement of cholesterol from extrahepatic tissues to 4. Do not mix fresh reagent with in-use reagents. the liver for disposal). ApoA-I also has a role in the activation of LCAT 5. CAUTION: This product requires the handling of human specimens. (lecithin-cholesterol acyltransferase).1-3 It is recommended that all human sourced materials be considered Measurement of ApoA-I is useful in predicting patients with high risk potentially infectious and be handled in accordance with the OSHA of (CAD). Levels of ApoA-I are inversely Standard on Bloodborne Pathogens.6 Biosafety Level 27 or other correlated with the risk of premature CAD. The relative proportion of appropriate biosafety practices8,9 should be used for materials that ApoB, a major component of VLDL and LDL, to ApoA is effective in contain or are suspected of containing infectious agents. differentiating individuals with or without ischemic heart disease. An 6. This product contains sodium azide. For a specific listing, refer to increased ApoB/ApoA ratio at a young age is potentially a marker for 4 the REAGENTS section of this package insert. Contact with acids CAD. liberates very toxic gas. This material and its container must be Elevated ApoA-I levels are seen in familial hyper-α-lipoproteinemia disposed of in a safe way. and weight reduction. Decreased levels are associated with various NOTE: Refer to Section 8 of the instrument-specific operations forms of familial and non-familial hypo-α-lipoproteinemia, Tangier manual for proper handling and disposal of reagents containing disease, premature CAD, hypertriglyceridemia, uncontrolled diabetes, sodium azide. hepatocellular disorders, cholestasis, nephrotic syndrome, and chronic For product not classified as dangerous per European Directive renal failure.5 1999/45/EC as amended, safety data sheet available for professional PRINCIPLES OF PROCEDURE user on request. The ApoA assay is an immunoturbidimetric procedure that measures SPECIMEN COLLECTION AND HANDLING increasing sample turbidity caused by the formation of insoluble immune complexes when antibody to ApoA-I is added to the sample. Suitable Specimens Sample containing ApoA-I is incubated with a buffer ( ) and a sample Serum and plasma are acceptable specimens. Fasting sample blank determination is performed prior to the addition of ApoA-I antibody (≥ 12 hours) is recommended.4,5 ( ). In the presence of an appropriate antibody in excess, the ApoA-I • Serum: Use serum collected by standard venipuncture techniques concentration is measured as a function of turbidity. into glass or plastic tubes with or without gel barriers. Ensure Methodology: Immunoturbidimetric complete clot formation has taken place prior to centrifugation. Separate serum from red blood cells or gel as soon after collection REAGENTS as possible. Reagent Kit Some specimens, especially those from patients receiving anticoagulant or thrombolytic therapy, may take longer to complete 9D92 Apolipoprotein A1 is supplied as a liquid, ready-to-use, their clotting processes. Fibrin clots may subsequently form in these two-reagent kit which contains: sera and the clots could cause erroneous test results. 3 x 21 mL • Plasma: Use plasma collected by standard venipuncture techniques 3 x 9 mL into glass or plastic tubes. Acceptable anticoagulants are lithium heparin (with or without gel barrier), sodium heparin, and EDTA. Estimated tests per kit: 243 Ensure centrifugation is adequate to remove platelets. Separate Calculation is based on the minimum reagent fill volume per kit. plasma from red blood cells or gel as soon after collection as possible. Reactive Ingredients Concentration TRIS 100 mmol/L Refer to the specimen collection tube manufacturer’s instructions for processing and handling requirements. Polyethylene Glycol 35 g/L For total sample volume requirements, refer to the instrument-specific Sodium Azide 0.1% ASSAY PARAMETERS section of this package insert and Section 5 of Anti-human apolipoprotein A1 goat serum 50% the instrument-specific operations manual. TRIS 100 mmol/L Sodium Azide 0.1%

2 SPECIMEN COLLECTION AND HANDLING (Continued) CALIBRATION Specimen Storage The linear high field of the assay parameters must be edited to the concentration of the highest calibrator specified in the value sheet. Serum and Plasma: Analyze fresh specimens if possible. Repeated freeze/thaw cycles should be avoided to minimize potential protein Calibration is stable for approximately 57 days (1,368 hours) and is degradation. required with each change in reagent lot number. Verify calibration with at least three levels of controls according to the established quality Temperature Maximum Bibliographic control requirements for your laboratory. If control results fall outside Storage Reference acceptable ranges, recalibration may be necessary. A multi-point calibration (Linear) curve is generated using Apo A1/Apo B 2 to 8°C 3 days 10, 11 Calibrator. -20°C 2 months 10 For a detailed description of how to calibrate an assay, refer to Guder et al.10 suggest storage of frozen specimens at -20°C for Section 6 of the instrument-specific operations manual. no longer than the time interval cited above. However, limitations For information on calibrator standardization, refer to the Apo A1/Apo B of laboratory equipment make it necessary in practice for clinical Calibrator package insert. laboratories to establish a range around -20°C for specimen storage. This temperature range may be established from either the freezer QUALITY CONTROL manufacturer’s specifications or your laboratory standard operating The following is the recommendation of Abbott Laboratories for quality procedure(s) for specimen storage. control. As appropriate, refer to your laboratory standard operating NOTE: Stored specimens must be inspected for particulates. If present, procedure(s) and/or quality assurance plan for additional quality control mix and centrifuge the specimen to remove particulates prior to testing. requirements and potential corrective actions. • Three levels of quality control are to be run every 24 hours. PROCEDURE • Run three levels of quality control with each cartridge change. Materials Provided • If more frequent control monitoring is required, follow the established 9D92 Apolipoprotein A1 Reagent Kit quality control procedures for your laboratory. • If quality control results do not meet the acceptance criteria Materials Required but not Provided defined by your laboratory, patient values may be suspect. Follow • 6E54 Apo A1/Apo B Calibrator the established quality control procedures for your laboratory. 1 x 1 mL Recalibration may be necessary. 2 x 2 mL • Review quality control results and acceptance criteria following a change of reagent or calibrator lot. • Control Material • Saline (0.85% to 0.90% NaCl) for specimens that require dilution RESULTS Assay Procedure Refer to the instrument-specific operations manual for information on For a detailed description of how to run an assay, refer to Section 5 of results calculations. the instrument-specific operations manual. • ARCHITECT System Operations Manual—Appendix C Specimen Dilution Procedures • AEROSET System Operations Manual—Appendix A The ARCHITECT c Systems and the AEROSET System have automatic Representative performance data are given in the EXPECTED VALUES dilution features; refer to Section 2 of the instrument-specific operations and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this manual for additional information. package insert. Results obtained in individual laboratories may vary. Serum and Plasma: Specimens with ApoA1 values exceeding the LIMITATIONS OF THE PROCEDURE highest calibrator are flagged and may be diluted by following either the Automated Dilution Protocol or the Manual Dilution Procedure. Refer to the SPECIMEN COLLECTION AND HANDLING and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this package insert. Automated Dilution Protocol The performance characteristics of ApoA on an analyzer other than the If using the Automated Dilution Protocol, the system performs a 1:2 ARCHITECT c Systems or the AEROSET System must be validated and dilution of the specimen and automatically corrects the concentration by verified. multiplying the result by the appropriate dilution factor. Grossly lipemic specimens may cause nonspecific light scattering. Manual Dilution Procedure These specimens can be diluted with isotonic saline to reduce the nonspecific light scattering. Refer to the Specimen Dilution Procedures Manual dilutions should be performed as follows: section of this package insert for specimen dilution instructions. • Use saline (0.85% to 0.90% NaCl) to dilute the sample. Samples containing paraproteins (abnormal monoclonal antibodies) • The operator must enter the dilution factor in the patient or control may interfere with test results. Samples with elevated total order screen. The system uses this dilution factor to automatically protein concentrations or samples from patients with suspected correct the concentration by multiplying the result by the entered paraproteinemia can be screened using other laboratory methods such factor. as protein electrophoresis.12 • If the operator does not enter the dilution factor, the result must be multiplied by the appropriate dilution factor before reporting the result. NOTE: If a diluted sample result is flagged indicating it is less than the linear low limit, do not report the result. Rerun using an appropriate dilution. For detailed information on ordering dilutions, refer to Section 5 of the instrument-specific operations manual. The patient result flag “>” (ARCHITECT c Systems) and the EXT and LH result error codes (AEROSET) may indicate antigen excess. Dilute sample and rerun. Samples were tested for antigen excess up to 605.4 mg/dL (6.054 g/L).

3 EXPECTED VALUES SPECIFIC PERFORMANCE CHARACTERISTICS Reference Range (Continued) Serum/Plasma13 Interfering Substances Interference studies were conducted using CLSI protocol NCCLS Range* (mg/dL) Range* (g/L) EP7-P.18 Interference effects were assessed by Dose Response and 0 to 1 year Paired Difference methods, at the medical decision level of the analyte. Male 61 to 164 0.61 to 1.64 Interfering Interferent Concentration N Target Observed Female 59 to 169 0.59 to 1.69 Substance (mg/dL) (% of Target) > 1 to 12 years 15 mg/dL (257 μmol/L) 4 143.4 93.5 Bilirubin Male 93 to 172 0.93 to 1.72 30 mg/dL (513 μmol/L) 4 143.4 85.6 Female 86 to 179 0.86 to 1.79 1,000 mg/dL (10.0 g/L) 4 152.3 92.0 Hemoglobin > 12 to 60 years 2,000 mg/dL (20.0 g/L) 4 152.3 84.3 Male 95 to 186 0.95 to 1.86 Human 500 mg/dL (5.7 mmol/L) 4 165.2 90.6 Female 101 to 223 1.01 to 2.23 750 mg/dL (8.5 mmol/L) 4 165.2 86.0 > 60 years 1,000 mg/dL (10.0 g/L) 4 150.5 100.7 Intralipid Male 73 to 186 0.73 to 1.86 2,000 mg/dL (20.0 g/L) 4 150.5 99.9 Female 91 to 224 0.91 to 2.24 Bilirubin solutions at the above concentrations were prepared by * Caucasian values (African-American values are 5 to 10 mg/dL addition of a bilirubin stock to human serum pools. Hemoglobin 14-16 higher. ) solutions at the above concentrations were prepared by addition Reference ranges are based on a 90% confidence interval. of hemolysate to human serum pools. Human triglyceride solutions at the above concentrations were prepared by mixing an elevated To convert results from mg/dL to g/L, multiply mg/dL by 0.01. triglyceride human serum pool with a normal triglyceride human serum A study was conducted using 121 serum samples from volunteers. pool. Intralipid solutions at the above concentrations were prepared by Data were analyzed as described by Clinical and Laboratory Standards addition of Intralipid to human serum pools. Institute (CLSI) protocol NCCLS C28-A.17 From this study, 95% of all specimens fell within 107.54 to 218.79 mg/dL (1.07 to 2.19 g/L), with Precision samples ranging from 100.68 to 236.17 mg/dL (1.01 to 2.36 g/L). The imprecision of the ApoA assay is ≤ 4.9% Total CV. Representative 19 20 It is recommended that each laboratory determine its own reference data from studies using CLSI protocols NCCLS EP5-T2 and EP5-A range based upon its particular locale and population characteristics. are summarized below. SPECIFIC PERFORMANCE CHARACTERISTICS Control Level 1 Level 2 Level 3 N 808080 Reportable Range (Accuracy by Recovery) The ApoA assay reportable range is from 16 mg/dL (0.16 g/L) to the Mean (mg/dL) 66.2 188.3 208.8 highest calibrator concentration. Human serum containing a known SD 1.27 2.17 2.46 concentration of ApoA1 was diluted with saline and the resulting Within Run samples were analyzed. Observed mean results across the reportable %CV 1.9 1.2 1.2 range were within 3 mg/dL (0.03 g/L) or 10%, whichever is greater, of SD 0.00 1.66 2.96 the target concentrations. Representative data are summarized below. Between Run %Recovery = (Observed Mean / Target Concentration) × 100 %CV 0.0 0.9 1.4 SD 1.68 4.31 0.00 Target Observed Delta* Percent (%) Between Day Concentration Mean (mg/dL) Recovery* %CV 2.5 2.3 0.0 (mg/dL) (mg/dL) SD 2.11 5.10 3.85 Total 2 . 6 1. 4 -1. 1 5 6. 4 %CV 3.2 2.7 1.8 10.2 7.6 -2.6 74.3 17.1 15.1 -2.0 88.3 Method Comparison 34.1 31.4 -2.8 91.9 Correlation studies were performed using CLSI protocol NCCLS 21 68.3 64.5 -3.8 94.4 EP9-A. 136.6 142.0 5.4 104.0 Serum results from the ApoA assay on the AEROSET System were compared with those from a commercially available immunoturbidimetric 204.9 206.8 1.9 100.9 methodology. 273.2 271.2 -1.9 99.3 Serum results from the ApoA assay on an ARCHITECT c System were 341.4 340.7 -0.7 99.8 compared with the ApoA assay on an AEROSET System. * Delta and %Recovery were calculated prior to rounding Target AEROSET vs. ARCHITECT Concentration and Observed Mean values. Comparative Method vs. AEROSET Limit of Quantitation (LOQ) N8095 The LOQ for ApoA is ≤ 3 mg/dL (0.03 g/L). The LOQ is the analyte Y - Intercept 6.24 -5.59 concentration at which the CV = 20%. Performance studies produced an LOQ of 0.8 mg/dL (0.008 g/L). Correlation Coefficient 0.992 0.996 Slope 0.98 0.99 Mean %Bias 3.3 -4.8 Range (mg/dL) 26.9 to 196.8 5.8 to 335.2

4 BIBLIOGRAPHY 1. Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical Chemistry, 2nd ed. Philadelphia, PA: WB Saunders; 1994:1019–21. 2. Vélez-Carrasco W, Lichtenstein AH, Barrett PHR, et al. Human apolipoprotein A-I kinetics within triglyceride-rich lipoproteins and high density lipoproteins. Journal of Lipid Research 1999;40:1695–1700. 3. Tailleux A, Duriez P, Fruchart JC, et al. Apolipoprotein A-II, HDL metabolism and . Atherosclerosis [serial online] 2002;164:1–13. Available from Elsevier Science Ireland, Ltd. 4 Jacobs DS, DeMott WR, Grady HJ, et al. Laboratory Test Handbook, 4th ed. Hudson, OH: Lexi-Comp; 1996:82. 5. Tietz NW, editor. Clinical Guide to Laboratory Tests, 3rd ed. Philadelphia, PA: WB Saunders; 1995:68–9. 6. US Department of Labor, Occupational Safety and Health Administration. 29 CFR Part 1910.1030. Bloodborne Pathogens. 7. US Department of Health and Human Services. Biosafety in Microbiological and Biomedical Laboratories, 5th ed. Washington, DC: US Government Printing Office, January 2007. 8. World Health Organization. Laboratory Biosafety Manual, 3rd ed. Geneva: World Health Organization, 2004. 9. Sewell DL, Bove KE, Callihan DR, et al. Protection of Laboratory Workers from Occupationally Acquired Infections; Approved Guideline—Third Edition (M29-A3). Wayne, PA: Clinical and Laboratory Standards Institute, 2005. 10. Guder WG, da Fonseca-Wollheim F, Heil W, et al. The Quality of Diagnostic Samples. Darmstadt, Germany: GIT Verlag; 2001:18–9. 11. US Pharmacopeial Convention, Inc. General notices. In: US Pharmacopeia National Formulary, 1995 ed (USP 23/NF 18). Rockville, MD: The US Pharmacopeial Convention, Inc; 1994:11. 12. Ledue TB, Collins MF, Ritchie RF. Development of immunoturbidimetric assays for fourteen human serum on the Hitachi 912. Clin Chem Lab Med 2002;40(5):520–8. 13. Ritchie RF, editor. Serum Proteins in Clinical Medicine, Vol 1. AACC, 1996:12.01-7. 14. Tietz NW, editor. Clinical Guide to Laboratory Tests, 3rd ed. Philadelphia, PA: WB Saunders; 1995:336. 15. Rubins HB, Robins SJ, Collins D, et al. Distribution of lipids in 8,500 men with coronary artery disease. Department of Veterans Affairs HDL Intervention Trial Study Group. Am J Cardiol 1995;15:1196–201. 16. Brown SA, Hutchinson R, Morrisett J, et al. Plasma lipid, lipoprotein cholesterol, and apoprotein distributions in selected US communities. The atherosclerosis risk in communities (ARIC) study. Arterioscler Thromb 1993;13:1139–58. 17. Sasse EA, Aziz KJ, Harris EK, et al. How to Define and Determine Reference Intervals in the Clinical Laboratory; Approved Guideline (C28-A). Villanova, PA: The National Committee for Clinical Laboratory Standards, 1995. 18. Powers DM, Boyd JC, Glick MR, et al. Interference Testing in Clinical Chemistry; Proposed Guideline (EP7-P). Villanova, PA: The National Committee for Clinical Laboratory Standards, 1986. 19. Kennedy JW, Carey RN, Coolen RB, et al. Evaluation of Precision Performance of Clinical Chemistry Devices—Second Edition; Tentative Guideline (EP5-T2). Villanova, PA: The National Committee for Clinical Laboratory Standards, 1992. 20. Kennedy JW, Carey RN, Coolen RB, et al. Evaluation of Precision Performance of Clinical Chemistry Devices; Approved Guideline (EP5-A). Wayne, PA: The National Committee for Clinical Laboratory Standards, 1999. 21. Kennedy JW, Carey RN, Coolen RB, et al. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline (EP9-A). Wayne, PA: The National Committee for Clinical Laboratory Standards, 1995. TRADEMARKS The ARCHITECT c System family of instruments consists of c 4000, c 8000, and c 16000 instruments. AEROSET, ARCHITECT, c 4000, c 8000, c 16000, c System, and SmartWash are trademarks of Abbott Laboratories in various jurisdictions. All trademarks are property of their respective owners.

Abbott Laboratories ABBOTT Abbott Park, IL 60064 USA Max-Planck-Ring 2 65205 Wiesbaden Germany 304511/R5 +49-6122-580

5 ARCHITECT c SYSTEMS ASSAY PARAMETERS

Apolipoprotein A1 Serum/Plasma—Conventional and SI Units Configure assay parameters — General Configure assay parameters — SmartWash ● General о Calibration о SmartWash о Results о Interpretation о General о Calibration ● SmartWash о Results о Interpretation Assay: ApoA Type: Photometric Version: † Assay: ApoA Number: 1053 COMPONENT REAGENT / ASSAY WASH Volume Replicates ● Reaction definition о Reagent / Sample о Validity checks R1 DIG00 Detergent A 345 1 Reaction mode: End up R1 AMIK9 Detergent A 345 1 Primary Secondary Read times R1 VANCO Detergent A 345 1 Wavelength: 804 / None Main: 31 – 33 R1 GENT9 Detergent A 345 1 Last required read: 33 R1 TOBRA Detergent A 345 1 Absorbance range: ___ – ___ Color correction: ___ – ___ R1 DGT0B Detergent A 345 1 Sample blank type: Self Blank: 14 – 16 R2 DIG00 Detergent A 345 1 R2 AMIK9 Detergent A 345 1 о Reaction definition ● Reagent / Sample о Validity checks R2 VANCO Detergent A 345 1 R1 R2 R2 GENT9 Detergent A 345 1 Reagent: APOA0 Reagent volume: 200 67 R2 TOBRA Detergent A 345 1 Diluent: Saline Water volume: ______R2 DGT0B Detergent A 345 1 Diluent dispense mode: Type 0 Dispense mode: Type 0 Type 0 Cuvette Trig 10% Detergent B 345 Diluted Default Dilution name Sample sample Diluent Water Dilution factor dilution STANDARD : 2.0 ______= 1:1.00 ● Apolipoprotein A1 Serum/Plasma—Conventional Units 1:2 : 25.0 4.0 75 ___ = 1:2.01 о ______: ______= о Configure assay parameters — Results о General о Calibration о SmartWash ● Results о Interpretation о Reaction definition о Reagent / Sample ● Validity checks Assay: ApoA Assay number: 1053 Reaction check: None Dilution default range: Result units: mg/dL Low-Linearity: 16 ‡‡ Maximum absorbance variation: ___ High-Linearity: 310 Gender and age specific ranges:* GENDER AGE (UNITS) NORMAL** EXTREME Configure assay parameters — Calibration Male 0 – 130 (Y) 95 – 186 о General ● Calibration о SmartWash о Results о Interpretation Female 0 – 130 (Y) 101 – 223 Assay: ApoA Calibration method: Linear Either 0 – 130 (Y) 95 – 223 ● Calibrators о Volumes о Intervals о Validity checks Calibrator set: Calibrator level: Concentration: Configure result units APO Blank: Water 0†† Assay: ApoA Cal 1: APO1 ‡ Version: † Replicates: 3 [Range 1 – 3] Cal 2: APO2 ‡ Result units: mg/dL Cal 3: APO3 ‡ Decimal places: 0 [Range 0 – 4] Cal 4: APO4 ‡ Correlation factor: 1.0000 Cal 5: APO5 ‡ Intercept: 0.0000

Apolipoprotein A1 Serum/Plasma—SI Units о Calibrators ● Volumes о Intervals о Validity checks Calibrator: APO Diluted Configure assay parameters — Results Calibrator level Sample sample Diluent Water Blank: Water 2.0 ______о General о Calibration о SmartWash ● Results о Interpretation Cal 1: APO1 2.0 ______Assay: ApoA Assay number: 1053 Cal 2: APO2 2.0 ______Dilution default range: Result units: g/L Low-Linearity: 0.16 Cal 3: APO3 2.0 ______‡‡ Cal 4: APO4 2.0 ______High-Linearity: 3.10 Cal 5: APO5 2.0 ______Gender and age specific ranges:* GENDER AGE (UNITS) NORMAL** EXTREME Male 0 – 130 (Y) 0.95 – 1.86 о Calibrators о Volumes ● Intervals о Validity checks Female 0 – 130 (Y) 1.01 – 2.23 Calibration intervals: Either 0 – 130 (Y) 0.95 – 2.23 Full interval: 1368 (hours) Calibration type: Adjust type: None Configure result units Assay: ApoA о Calibrators о Volumes о Intervals ● Validity checks Version: † Blank absorbance range: _____ – _____ Result units: g/L Span: Blank – Blank Decimal places: 2 [Range 0 – 4] Span absorbance range: _____ – _____ Correlation factor: 1.0000 Expected cal factor: 0.00 Intercept: 0.0000 Expected cal factor tolerance %: 0 * User defined. ** Reference range is from > 12 years to 60 years of age. † Due to differences in instrument systems and unit configurations, version numbers may vary. †† Displays the number of decimal places defined in the decimal places parameter field. ‡ Refer to the concentration specified on calibrator labeling or value sheet. In ARCHITECT software version 5.00 and above, these values are defined on the Configure calibrator screen. ‡‡ Edit to highest calibrator concentration specified in the calibrator value sheet. 6 AEROSET SYSTEM ASSAY PARAMETERS

Apolipoprotein A1 Serum/Plasma—Conventional Units Apolipoprotein A1 Serum/Plasma—SI Units Assay Configuration: Outline Page Assay Configuration: Outline Page Assay Name Assay # Line Assay Name Assay # Line ApoA 53 A-Line ApoA 53 A-Line Quantitative Ranges Quantitative Ranges Min Text Min Panic-L L-Reference-H** Panic-H Max Max Text Min Text Min Panic-L L-Reference-H** Panic-H Max Max Text * 0.0* 0.0 95 223 0.0 0.0* * * 0.0* 0.0 0.95 2.23 0.0 0.0* * 16 L-Linear Range-H 310‡‡ 0.16 L-Linear Range-H 3.10‡‡ Reference Ranges* Reference Ranges* Age Male Female Age Male Female 0 Year 95 – 186 101 – 223 0 Year 0.95 – 1.86 1.01 – 2.23 0.0 – 0.0 0.0 – 0.0 0.0 – 0.0 0.0 – 0.0 0 Year 0 Year 0.0 – 0.0 0.0 – 0.0 0.0 – 0.0 0.0 – 0.0 0 Year 0.0 – 0.0 0.0 – 0.0 0 Year 0.0 – 0.0 0.0 – 0.0 Qualitative Ranges N/A Qualitative Ranges N/A

Assay Configuration: Base Page Assay Configuration: Base Page Reaction Mode Wavelength-Prim/Sec Read time-Main/Flex AbsMaxVar Reaction Mode Wavelength-Prim/Sec Read time-Main/Flex AbsMaxVar END UP 804 / ___ 31 – 33 / 0 – 0 0.0 END UP 804 / ___ 31 – 33 / 0 – 0 0.0 Sample Blank Test Blank Read Time Abs Window Abs Limits Sample Blank Test Blank Read Time Abs Window Abs Limits ApoA ( 53 ) 14 – 16 0 – 0 0.0 – 0.0 ApoA ( 53 ) 14 – 16 0 – 0 0.0 – 0.0 S.Vol DS.Vol D.Vol W.Vol S.Vol DS.Vol D.Vol W.Vol Standard 2.0 0.0 0 0 Rgt Name/Pos Standard 2.0 0.0 0 0 Rgt Name/Pos Dil 1 25.0 4.0 75 0 Diluent: DILUENT C–10* Dil 1 25.0 4.0 75 0 Diluent: DILUENT C–10* Dil 2 2.0 0.0 0 0 Type# 0 Dil 2 2.0 0.0 0 0 Type# 0 Rgt Name/Pos R.Vol W.Vol Type# Rgt Name/Pos R.Vol W.Vol Type# Reagent 1 APOA011 – ___* 200 0 0 Reagent 1 APOA011 – ___* 200 0 0 Reagent 2 APOA012 – ___* 67 0 0 Reagent 2 APOA012 – ___* 67 0 0 Reaction Check Read Time – A/B Range Minimum Reaction Check Read Time – A/B Range Minimum ______1 – 1 / 1 – 1 0.0 – 0.0 0.0 ______1 – 1 / 1 – 1 0.0 – 0.0 0.0 Factor/Intercept Decimal Places Units Factor/Intercept Decimal Places Units 1.0 / 0.0 0 mg/dL 1.0 / 0.0 2 g/L

Assay Configuration: Calibration Page Assay Configuration: Calibration Page Calib Mode Interval (H) Calib Mode Interval (H) Linear 1368 Linear 1368 Blank/Calib Replicates Extrapolation % Span Span Abs Range Blank/Calib Replicates Extrapolation % Span Span Abs Range 3 / 3 1 BLK – 1 0.0 – 0.0 3 / 3 1 BLK – 1 0.0 – 0.0 Sample S.Vol DS.Vol D.Vol W.Vol BLK Abs Range Sample S.Vol DS.Vol D.Vol W.Vol BLK Abs Range BLK Water 2.0 0.0 0 0 0.0 – 0.0 BLK Water 2.0 0.0 0 0 0.0 – 0.0 C1 APO 1 2.0 0.0 0 0 Cal Deviation C1 APO 1 2.0 0.0 0 0 Cal Deviation C2 APO 2 2.0 0.0 0 0 0.0 C2 APO 2 2.0 0.0 0 0 0.0 C3 APO 3 2.0 0.0 0 0 FAC Limit (%) C3 APO 3 2.0 0.0 0 0 FAC Limit (%) C4 APO 4 2.0 0.0 0 0 10 C4 APO 4 2.0 0.0 0 0 10 C5 APO 5 2.0 0.0 0 0 C5 APO 5 2.0 0.0 0 0

Assay Configuration: SmartWash Page Assay Configuration: SmartWash Page Rgt Probe Rgt Probe Reagent Wash Vol Reagent Wash Vol DIG0051 AlkW 345 DIG0051 AlkW 345 DIG0012 AlkW 345 DIG0012 AlkW 345 AMIK941 AlkW 345 AMIK941 AlkW 345 AMIK942 AlkW 345 AMIK942 AlkW 345 VANCO51 AlkW 345 VANCO51 AlkW 345 VANCO52 AlkW 345 VANCO52 AlkW 345 TOBRA41 AlkW 345 TOBRA41 AlkW 345 TOBRA42 AlkW 345 TOBRA42 AlkW 345 DGT0B11 AlkW 345 DGT0B11 AlkW 345 DGT0B12 AlkW 345 DGT0B12 AlkW 345 Cuvette Cuvette Assay Name Wash Vol Assay Name Wash Vol ——— ——— Sample Probe Sample Probe Wash Wash — —

Refer to Assay Configuration in Section 2 of the AEROSET System Operations Manual for information regarding assay parameters. * User defined or instrument defined. ** Reference range is from > 12 years to 60 years of age. ‡‡ Edit to highest calibrator concentration specified in the calibrator value sheet.

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