Haptoglobin 9D91-20 30-3966/R4

HAPTOGLOBIN 9D91-20 30-3966/R4

HAPTOGLOBIN

This package insert contains information to run the Haptoglobin 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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Calibrators 1 through 5 Catalog number/List number

Concentration Serial number Authorized Representative in the Consult instructions for use European Community Ingredients Manufacturer

In vitro diagnostic medical device Temperature limitation

Batch code/Lot number Use by/Expiration date

Reagent 1

Reagent 2

1 NAME REAGENT HANDLING AND STORAGE HAPTOGLOBIN 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 Haptoglobin assay is used for the quantitation of haptoglobin in storage temperature to allow the bubbles to dissipate. To minimize 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 Haptoglobin is a protein synthesized in the liver that binds with the which could impact results. globin α-chains of hemoglobin A, F, S, or C. Haptoglobin does not bind methemoglobin, heme, or unusual forms of hemoglobin in which Reagent Storage the α-chain is missing. The haptoglobin-hemoglobin complex is rapidly Unopened reagents are stable until the expiration date when stored removed from circulation by the reticuloendothelial system to prevent/ at 2 to 8°C. minimize hemoglobin loss and to conserve iron.1 Reagent onboard stability is approximately 57 days if quality control Indications for haptoglobin quantitation include: anemia or other results meet acceptance criteria. If quality control results do not meet indicators of possible hemolysis; pregnancy-induced hypertension; acceptance criteria, refer to the QUALITY CONTROL section of this transfusion reactions (assay of pre- and post-transfusion samples); package insert. assessment of acute phase response; and evaluation of changes in the 2 α2-region of serum protein electrophoresis. WARNINGS AND PRECAUTIONS Decreased levels of haptoglobin are most frequently associated with Precautions for Users conditions of increased intravascular hemolysis or hemoglobin turnover, such as hemolytic anemias, intravascular hemolytic transfusion 1. For in vitro diagnostic use. reactions, and malaria. Extravascular hemolysis does not usually affect 2. Do not use components beyond the expiration date. haptoglobin concentrations. Other conditions associated with decreased 3. Do not mix materials from different kit lot numbers. haptoglobin levels are congenital anhaptoglobinemia (ahaptoglobinemia) 4. Do not mix fresh reagent with in-use reagents. and severe liver diseases where protein synthesis is affected.1 5. This product contains sodium azide; for a specific listing, refer to the Mechanical heart valves or vigorous athletic activity where repetitive REAGENTS section. Contact with acids liberates very toxic gas. This erythrocytic mechanical trauma may occur can cause sustained lower material and its container must be disposed of in a safe way. 1,2 haptoglobin levels. NOTE: Refer to Section 8 of the instrument-specific operations Elevated levels of haptoglobin are most frequently associated with manual for proper handling and disposal of reagents containing acute phase reactions (i.e., involving tissue infection, surgery, trauma, sodium azide. or necrosis).3 Other conditions associated with elevated haptoglobin 2 6. CAUTION: This product requires the handling of human specimens. levels are corticosteroid therapy and biliary obstruction. It is recommended that all human sourced materials be considered potentially infectious and be handled in accordance with the OSHA PRINCIPLES OF PROCEDURE Standard on Bloodborne Pathogens.4 Biosafety Level 25 or other The Haptoglobin assay is an immunoturbidimetric procedure that appropriate biosafety practices6,7 should be used for materials that measures increasing sample turbidity caused by the formation of contain or are suspected of containing infectious agents. insoluble immune complexes when antibody to haptoglobin is added to the sample. Sample containing haptoglobin is incubated with a buffer For product not classified as dangerous per European Directive (Reagent 1) and a sample blank determination is performed prior to 1999/45/EC as amended, safety data sheet available for professional the addition of haptoglobin antibody (Reagent 2). In the presence of user on request. an appropriate antibody in excess, the haptoglobin concentration is measured as a function of turbidity. SPECIMEN COLLECTION AND HANDLING Methodology: Immunoturbidimetric Suitable Specimens Serum and plasma are acceptable specimens. REAGENTS • Serum: Use serum collected by standard venipuncture techniques Reagent Kit into glass or plastic tubes with or without gel barriers. Ensure 9D91 Haptoglobin is supplied as a liquid, ready-to-use, two-reagent complete clot formation has taken place prior to centrifugation. kit which contains: Separate serum from red blood cells or gel as soon after collection as possible. 3 x 17 mL Some specimens, especially those from patients receiving 3 x 7 mL anticoagulant or thrombolytic therapy, may take longer to complete Estimated tests per kit: 230 their clotting processes. Fibrin clots may subsequently form in these sera and the clots could cause erroneous test results. Calculation is based on the minimum reagent fill volume per kit. • Plasma: Use plasma collected by standard venipuncture techniques Reactive Ingredients Concentration into glass or plastic tubes. Acceptable anticoagulants are lithium heparin (with or without gel barrier), sodium heparin, and EDTA. TRIS 100 mmol/L Ensure centrifugation is adequate to remove platelets. Separate plasma from red blood cells or gel as soon after collection as Polyethylene Glycol 30 g/L possible. Sodium Azide 0.1% Refer to the specimen collection tube manufacturer’s instructions for Anti-human haptoglobin goat serum 50% processing and handling requirements. TRIS 100 mmol/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 the instrument-specific operations manual.

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 (Spline) curve is generated using Specific 2 to 8°C 1 week 8, 9 Proteins Multiconstituent Calibrator. -20°C 2 weeks 2 For a detailed description of how to calibrate an assay, refer to Section 6 of the instrument-specific operations manual. Teitz2 suggests storage of frozen specimens at -20°C for no longer than the time interval cited above. However, limitations of laboratory For information on calibrator standardization, refer to the Specific equipment make it necessary in practice for clinical laboratories to Proteins Multiconstituent Calibrator package insert. establish a range around -20°C for specimen storage. This temperature range may be established from either the freezer manufacturer’s QUALITY CONTROL specifications or your laboratory standard operating procedure(s) for The following is the recommendation of Abbott Laboratories for quality 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 • If more frequent control monitoring is required, follow the established Materials Provided quality control procedures for your laboratory. 9D91 Haptoglobin Reagent Kit • If quality control results do not meet the acceptance criteria defined by your laboratory, patient values may be suspect. Follow Materials Required but not Provided the established quality control procedures for your laboratory. • 1E78 Specific Proteins Multiconstituent Calibrator Recalibration may be necessary. 1 x 1 mL • Review quality control results and acceptance criteria following a • Control Material change of reagent or calibrator lot. • 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 haptoglobin values exceeding the highest calibrator are flagged and may be diluted using the Automated LIMITATIONS OF THE PROCEDURE 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 Haptoglobin on an analyzer other If using the Automated Dilution Protocol, the system performs a 1:2 or than the ARCHITECT c Systems or the AEROSET System must be 1:4 dilution of the specimen and automatically corrects the concentration validated and verified. by multiplying the result by the appropriate dilution factor. Samples containing paraproteins (abnormal monoclonal antibodies) Manual Dilution Procedure may interfere with test results. Samples with elevated total Manual dilutions should be performed as follows: protein concentrations or samples from patients with suspected paraproteinemia can be screened using other laboratory methods such • Use saline (0.85% to 0.90% NaCl) to dilute the sample. as protein electrophoresis.10 • The operator must enter the dilution factor in the patient or control Turbidity and particles in the samples can interfere with the assay. order screen. The system uses this dilution factor to automatically Therefore, particulate matter should be removed by centrifugation prior correct the concentration by multiplying the result by the entered to running the assay. factor. • 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 1,433 mg/dL (14.33 g/L).

3 EXPECTED VALUES SPECIFIC PERFORMANCE CHARACTERISTICS Reference Range (Continued) Serum/Plasma11 Interfering Substances Interference studies were conducted using CLSI protocol NCCLS Range* (mg/dL) Range* (g/L) EP7-P.13 Interference effects were assessed by Dose Response and 0 to 1 year Paired Difference methods, at the medical decision level of the analyte. Male 0 to 300 0.00 to 3.00 Interfering Interferent Concentration N Target Observed Female 0 to 235 0.00 to 2.35 Substance (mg/dL) (% of Target) > 1 to 12 years 30 mg/dL (513 μmol/L) 4 171.3 96.6 Bilirubin Male 3 to 270 0.03 to 2.70 60 mg/dL (1,026 μmol/L) 4 171.3 93.2 Female 11 to 220 0.11 to 2.20 1,000 mg/dL (10.0 g/L) 4 138.9 90.1 Hemoglobin > 12 to 60 years 2,000 mg/dL (20.0 g/L) 4 138.9 89.8 Male 14 to 258 0.14 to 2.58 Human 750 mg/dL (8.5 mmol/L) 4 175.7 103.2 Female 35 to 250 0.35 to 2.50 triglyceride 1,000 mg/dL (11.3 mmol/L) 4 175.7 102.6 > 60 years 1,000 mg/dL (10.0 g/L) 4 133.1 99.8 Intralipid Male 40 to 268 0.40 to 2.68 2,000 mg/dL (20.0 g/L) 4 133.1 97.1 Female 63 to 273 0.63 to 2.73 Bilirubin solutions at the above concentrations were prepared by addition of a bilirubin stock to human serum pools. Hemoglobin * Reference ranges are based on a 90% confidence interval. solutions at the above concentrations were prepared by addition To convert results from mg/dL to g/L, multiply mg/dL by 0.01. of hemolysate to human serum pools. Human triglyceride solutions at the above concentrations were prepared by mixing an elevated NOTE: The reportable range may not be suitable for the reference triglyceride human serum pool with a normal triglyceride human serum range of certain populations. An alternate method should be used for pool. Intralipid solutions at the above concentrations were prepared by samples with results below the reportable range. addition of Intralipid to human serum pools. A study was conducted using 121 serum samples from volunteers. Data were analyzed as described by Clinical and Laboratory Standards Precision 12 Institute (CLSI) protocol NCCLS C28-A. From this study, 95% of all The imprecision of the Haptoglobin assay is ≤ 6.0% Total CV. specimens fell within 31.62 to 213.16 mg/dL (0.32 to 2.13 g/L), with Representative data from studies using CLSI protocols NCCLS samples ranging from 16.24 to 241.31 mg/dL (0.16 to 2.41 g/L). EP5-T214 and EP5-A15 are summarized below. It is recommended that each laboratory determine its own reference range based upon its particular locale and population characteristics. Control Level 1 Level 2 Level 3 N 808080 SPECIFIC PERFORMANCE CHARACTERISTICS Mean (mg/dL) 59.4 122.7 182.3 Reportable Range (Accuracy by Recovery) SD 0.75 1.87 1.79 The Haptoglobin assay reportable range is from 8 mg/dL (0.08 g/L) to Within Run the highest calibrator concentration. Human serum containing a known %CV 1.3 1.5 1.0 concentration of haptoglobin was diluted with saline and the resulting samples were analyzed. Observed mean results across the reportable SD 0.72 1.13 2.84 Between Run range were within 4 mg/dL (0.04 g/L) or 10%, whichever is greater, of %CV 1.2 0.9 1.6 the target concentrations. Representative data are summarized below. SD 3.19 1.41 2.97 %Recovery = (Observed Mean / Target Concentration) × 100 Between Day Target Concentration Observed Mean Delta* Percent (%) %CV 5.4 1.2 1.6 (mg/dL) (mg/dL) (mg/dL) Recovery* SD 3.35 2.60 4.49 Total 3.4 3.8 0.4 110.8 %CV 5.6 2.1 2.5 7.9 7.6 -0.3 96.0 Method Comparison 13.2 13.3 0.1 100.7 Correlation studies were performed using CLSI protocol NCCLS EP9-A.16 26.3 25.5 -0.8 97.0 Serum results from the Haptoglobin assay on an AEROSET 52.6 49.8 -2.8 94.7 System were compared with those from a commercially available immunoturbidimetric methodology. 105.3 107.6 2.3 102.2 Serum results from the Haptoglobin assay on an ARCHITECT c System 157.9 163.8 5.9 103.7 were compared with the Haptoglobin assay on an AEROSET System. 210.6 225.2 14.6 107.0 AEROSET vs. ARCHITECT vs. 263.2 275.7 12.5 104.7 Comparative AEROSET Method * Delta and %Recovery were calculated prior to rounding Target Concentration and Observed Mean values. N4791 Limit of Quantitation (LOQ) Y - Intercept 0.25 -0.88 The LOQ for Haptoglobin is ≤ 4 mg/dL (0.04 g/L). The LOQ is the Correlation Coefficient 0.998 0.999 analyte concentration at which the CV = 20%. Performance studies Slope 1.03 0.98 produced an LOQ of 2.6 mg/dL (0.026 g/L). Mean %Bias 5.0 -4.1 Range (mg/dL) 1.8 to 276.8 13.4 to 279.0

4 BIBLIOGRAPHY 1. Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical Chemistry, 2nd ed. Philadelphia, PA: WB Saunders; 1994:707–8. 2. Tietz NW, editor. Clinical Guide to Laboratory Tests, 3rd ed. Philadelphia, PA: WB Saunders; 1995:306–9. 3. Henry JB. Clinical Diagnosis and Management by Laboratory Methods, 18th ed. Philadelphia, PA: WB Saunders; 1991:225–7. 4. US Department of Labor, Occupational Safety and Health Administration. 29 CFR Part 1910.1030. Occupational Exposure to Bloodborne Pathogens. 5. US Department of Health and Human Services. Biosafety in Microbiological and Biomedical Laboratories. 5th ed. Washington, DC: US Government Printing Office; January 2007. 6. World Health Organization. Laboratory Biosafety Manual. Geneva: World Health Organization, 2004. 7. 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. 8. Young DS. Effects of Preanalytical Variables on Clinical Laboratory Tests, 2nd ed. Washington, DC: AACC Press; 1997:4-497. 9. 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. 10. Ledue TB, Collins MF, Ritchie RF. Development of immunoturbidimetric assays for fourteen human serum proteins on the Hitachi 912. Clin Chem Lab Med 2002;40(5):520–8. 11. Ritchie RF, editor. Serum Proteins in Clinical Medicine, Vol 1. AACC, 1996:7.04-5. 12. 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. 13. 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. 14. 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. 15. 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. 16. 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 AEROSET and ARCHITECT are registered trademarks of Abbott Laboratories. c System is a trademark of Abbott Laboratories. All other trademarks, brands, product names, and trade names are the property of their respective companies.

5 ARCHITECT c SYSTEMS ASSAY PARAMETERS

Haptoglobin Serum/Plasma—Conventional and SI Units Configure assay parameters — SmartWash Configure assay parameters — General о General о Calibration ● SmartWash о Results о Interpretation General Calibration SmartWash Results Interpretation ● о о о о Assay: Hapt Assay: Hapt Type: Photometric Version: † Number: 1060 COMPONENT REAGENT / ASSAY WASH Volume Replicates Reaction definition Reagent / Sample Validity checks R1 DIG00 Detergent A 345 1 ● о о R1 AMIK9 Detergent A 345 1 Reaction mode: End up R1 VANCO Detergent A 345 1 Primary Secondary Read times R2 DIG00 Detergent A 345 1 Wavelength: 604 / None Main: 31 – 33 R2 AMIK9 Detergent A 345 1 Last required read: 33 R2 VANCO Detergent A 345 1 Absorbance range: ___ – ___ Color correction: ___ – ___ Cuvette Trig 10% Detergent B*** 345 Sample blank type: Self Blank: 14 – 16 *** Select “Detergent B” for software prior to Version 2.2.

о Reaction definition ● Reagent / Sample о Validity checks Haptoglobin Serum/Plasma—Conventional Units R1 R2 Reagent: HAPT0 Reagent volume: 167 47 Configure assay parameters — Results Diluent: Saline Water volume: ______о General о Calibration о SmartWash ● Results о Interpretation Diluent dispense mode: Type 0 Dispense mode: Type 0 Type 0 Assay: Hapt Result units: mg/dL Diluted Default Assay defaults: Dilution name Sample sample Diluent Water Dilution factor dilution Low-Linearity: 8 STANDARD : 2.0 ______= 1:1.00 ● High-Linearity: 250‡‡ 1:2 : 25.0 4.0 75 ___ = 1:2.02 о Gender and age specific ranges:* 1:4 : 25.0 2.0 75 ___ = 1:4.00 о GENDER AGE (UNITS) NORMAL** EXTREME Male 0 – 130 (Y) 14 – 258 Female 0 – 130 (Y) 35 – 250 о Reaction definition о Reagent / Sample ● Validity checks Reaction check: None Either 0 – 130 (Y) 14 – 258

Maximum absorbance variation: ___ Configure result units Configure assay parameters — Calibration Assay: Hapt о General ● Calibration о SmartWash о Results о Interpretation Version: † Assay: Hapt Calibration method: Spline Result units: mg/dL Decimal places: 0 [Range 0 – 4] ● Calibrators о Volumes о Intervals о Validity checks Correlation factor: 1.0000 Calibrator set: Calibrator level: Concentration: Intercept: 0.0000 SP Blank: Water 0†† Cal 1: SP1 ‡ Replicates: 3 [Range 1 – 3] Cal 2: SP2 ‡ Cal 3: SP3 ‡ Haptoglobin Serum/Plasma—SI Units Cal 4: SP4 ‡ Cal 5: SP5 ‡ Configure assay parameters — Results о General о Calibration о SmartWash ● Results о Interpretation о Calibrators ● Volumes о Intervals о Validity checks Assay: Hapt Result units: g/L Calibrator: SP Diluted Assay defaults: Calibrator level Sample sample Diluent Water Low-Linearity: 0.08 Blank: Water 2.0 ______High-Linearity: 2.50‡‡ Cal 1: SP1 2.0 ______Gender and age specific ranges:* Cal 2: SP2 2.0 ______GENDER AGE (UNITS) NORMAL** EXTREME Cal 3: SP3 2.0 ______Male 0 – 130 (Y) 0.14 – 2.58 Cal 4: SP4 2.0 ______Female 0 – 130 (Y) 0.35 – 2.50 Cal 5: SP5 2.0 ______Either 0 – 130 (Y) 0.14 – 2.58

о Calibrators о Volumes ● Intervals о Validity checks Calibration intervals: Configure result units Full interval: 1368 (hours) Assay: Hapt Calibration type: Version: † Adjust type: None Result units: g/L Decimal places: 2 [Range 0 – 4] о Calibrators о Volumes о Intervals ● Validity checks Correlation factor: 1.0000 Blank absorbance range: _____ – _____ Intercept: 0.0000 Span: Blank – Blank Span absorbance range: _____ – _____ Expected cal factor: 0.00 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. ‡‡ Edit to highest calibrator concentration specified in the calibrator value sheet. ‡ Refer to concentration specified on calibrator labeling or value sheet. 6 AEROSET SYSTEM ASSAY PARAMETERS

Haptoglobin Serum/Plasma—Conventional Units Haptoglobin Serum/Plasma—SI Units

Assay Configuration: Outline Page Assay Configuration: Outline Page Assay Name Assay # Line Assay Name Assay # Line Hapt 60 A-Line Hapt 60 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 14 258 0.0 0.0* * * 0.0* 0.0 0.14 2.58 0.0 0.0* * 8 L-Linear Range-H 250*** 0.08 L-Linear Range-H 2.50*** Reference Ranges** Reference Ranges** Age Male Female Age Male Female 0 Year 14 – 258 35 – 250 0 Year 0.14 – 2.58 0.35 – 2.50 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 604 / ___ 31 – 33 / 0 – 0 0.0 END UP 604 / ___ 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 Hapt ( 60 ) 14 – 16 0 – 0 0.0 – 0.0 Hapt ( 60 ) 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 25.0 2.0 75 0 Type# 0 Dil 2 25.0 2.0 75 0 Type# 0 Rgt Name/Pos R.Vol W.Vol Type# Rgt Name/Pos R.Vol W.Vol Type# Reagent 1 HAPT011 – ___* 167 0 0 Reagent 1 HAPT011 – ___* 167 0 0 Reagent 2 HAPT012 – ___* 47 0 0 Reagent 2 HAPT012 – ___* 47 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) Spline 1368 Spline 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 SP 1 2.0 0.0 0 0 Cal Deviation C1 SP 1 2.0 0.0 0 0 Cal Deviation C2 SP 2 2.0 0.0 0 0 0.0 C2 SP 2 2.0 0.0 0 0 0.0 C3 SP 3 2.0 0.0 0 0 FAC Limit (%) C3 SP 3 2.0 0.0 0 0 FAC Limit (%) C4 SP 4 2.0 0.0 0 0 10 C4 SP 4 2.0 0.0 0 0 10 C5 SP 5 2.0 0.0 0 0 C5 SP 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 AMIK941 Water 345 AMIK941 Water 345 AMIK942 Water 345 AMIK942 Water 345 VANCO51 AlkW 345 VANCO51 AlkW 345 VANCO52 AlkW 345 VANCO52 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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