IMMULITE 2000; HCG CALIBRATION VERIFICATION MATERIAL, INSULIN CALIBRATION VERIFICATION MATERIAL, PYRILINKS-D CALIBRATION

K133128 · Siemens Healthcare Diagnostics, Inc. · JJX · Oct 22, 2013 · Clinical Chemistry

Device Facts

• Record ID: K133128
• Device Name: IMMULITE 2000; HCG CALIBRATION VERIFICATION MATERIAL, INSULIN CALIBRATION VERIFICATION MATERIAL, PYRILINKS-D CALIBRATION
• Applicant: Siemens Healthcare Diagnostics, Inc.
• Product Code: JJX · Clinical Chemistry
• Decision Date: Oct 22, 2013
• Decision: SESE
• Submission Type: Abbreviated
• Regulation: 21 CFR 862.1660
• Device Class: Class 1

Intended Use

The IMMULITE® HCG Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE HCG assay on the IMMULITE 2000 systems. The IMMULITE® Insulin Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE Insulin assay on the IMMULITE 2000 systems. The IMMULITE® Pyrilinks-D Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE Pyrilinks-D assay on the IMMULITE 2000 systems. The IMMULITE® Homocysteine Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE Homocysteine assay on the IMMULITE 2000 systems. The IMMULITE® Growth Hormone Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE Growth Hormone assay on the IMMULITE 2000 systems.

Device Story

The IMMULITE 2000 Calibration Verification Materials (CVMs) are multi-level, analyte-specific control materials used to verify the calibration and reportable range of corresponding IMMULITE 2000 assays. The CVMs are prepared by spiking specific analytes (HCG, Insulin, Pyrilinks-D, Homocysteine, or Growth Hormone) into various matrices (human serum, equine serum, or phosphoric acid/sodium chloride). The device is used in clinical laboratory settings by trained personnel. The healthcare provider runs the CVMs on the IMMULITE 2000 system; the system generates dose values based on the reference calibrator curve. The provider compares these recovered values against established target ranges to verify assay calibration. If results fall within acceptable limits, the assay is considered calibrated; otherwise, corrective action is required. This process ensures the accuracy and reliability of patient sample testing, supporting clinical decision-making.

Clinical Evidence

Bench testing only. Stability studies validated shelf life (up to 6 years depending on analyte) and performance against acceptance criteria (±8% to ±15% of assigned dose). Value assignment was validated using 15-27 replicates across multiple systems and reagent lots, comparing recovered values to target means and 2SD ranges. No clinical patient outcome data provided.

Technological Characteristics

Multi-level (4 levels) liquid or lyophilized control materials. Matrices include human serum, equine serum, or phosphoric acid/sodium chloride with preservatives. Analyte-specific. Storage conditions vary (≤-20°C or 2-8°C). Traceable to WHO International Standards or internal gravimetric standards. Designed for use on IMMULITE 2000 immunoassay systems.

Indications for Use

Indicated for in vitro diagnostic use to verify calibration of specific IMMULITE 2000 assays (HCG, Insulin, Pyrilinks-D, Homocysteine, Growth Hormone) on IMMULITE 2000 systems. For prescription use only.

Regulatory Classification

Identification

A quality control material (assayed and unassayed) for clinical chemistry is a device intended for medical purposes for use in a test system to estimate test precision and to detect systematic analytical deviations that may arise from reagent or analytical instrument variation. A quality control material (assayed and unassayed) may be used for proficiency testing in interlaboratory surveys. This generic type of device includes controls (assayed and unassayed) for blood gases, electrolytes, enzymes, multianalytes (all kinds), single (specified) analytes, or urinalysis controls.

Submission Summary (Full Text)

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### Section 006: 510(k) Summary

#### 510(k) Summary

Introduction: According to the requirements of 21 CFR 807.92, the following information provides sufficient details to understand the basis for determination of substantial equivalence.

#### The assigned 510(k) Number: _k133128

...

- 1. Submitter
Mailing Address:

Contact Person:

Phone Number: Fax Number: E-mail Address: Date Prepared:

2. Device Name Proprietary Name: Measurand: Type of Test:

> Regulation Section: Classification: Products Code: Panel:

3. Predicate Device Name Predicate 510(k) No:

4. Device Description:

- 5. Intended Use: Indication for Use:
Siemens Healthcare Diagnostics Inc. 511 Benedict Avenue Tarrytown, NY 10591

:

Asha Gartland Technical Regulatory Affairs Specialist (914)-524-3257 (914)-524-2101 [e-mail] October 16, 2013

OCT 22 2013

#### IMMULITE® 2000 HCG Calibration Verification Material Quality Control materials for IMMULITE® 2000 HCG assay Calibration Verification Material (CVM) for IMMULITE® 2000 HCG assav 21 CFR 862.1660, Quality Control Material Class I Reserved JJX - Single (Specified) Analyte Controls (Assayed and Unassayed) Clinical Chemistry (75)

Elecsys HCG CalCheck 5 K092168

The Calibration Verification Material (CVM) contains One set of four vials each 3mL. CVM1 contains HCG-free human serum with preservatives. CVM2. CVM3 and CVM4 contain HCG added to HCG-free human serum respectively, with preservatives.

See Indications for Use Statement below:

The IMMULITE® HCG Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE HCG assay on the IMMULITE 2000 systems.

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- Special Conditions for Use Statement(s): Special Instrument Requirements:
- 6. Technological Characteristics and Substantial Equivalence Comparison with Predicate:

For prescription use only

IMMULITE® 2000 Systems

A comparison of the device features, intended use, and other information demonstrates that the IMMULITE® 2000 HCG Calibration Verification Material (CVM) is substantially equivalent to the predicate device as summarized in Table 1.

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### Table 1: Substantial Equivalence Comparison

| SIMILARITIES | | |
|-----------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | Candidate Device<br>IMMULITE 2000 HCG CVM | Predicate Device<br>Elecsys HCG CalCheck 5 |
| Intended<br>Use | The IMMULITE® HCG Calibration<br>Verification Material (CVM) is for in<br>vitro diagnostic use in the verification of<br>calibration of the IMMULITE HCG assay<br>on the IMMULITE 2000 systems. | The Elecsys HCG CalCheck 5 is an<br>assayed control for use in calibration<br>verification and for use in the<br>verification of the assay range<br>established by the Elecsys HCG+B<br>reagent and Elecsys HCG STAT reagent<br>on the indicated Elecsys and cobas e<br>immunoassay analyzers. |
| Analyte | HCG | Same |
| Matrix | Human Serum with preservatives | Same |
| Stability | Stable unopened until the expiration date | Same |

| | DIFFERENCES | |
|---------|-------------------------------------------|--------------------------------------------|
| | Candidate Device<br>IMMULITE 2000 HCG CVM | Predicate Device<br>Elecsys HCG CalCheck 5 |
| Form | Liquid | Lyophilized |
| Levels | 4 | 5 |
| Storage | ≤20°C | 2-8°C |
| Use | Single Use Only | Not For Single Use |

.

.

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#### 7. Non-Clinical Performance Testing

Performance testing has been carried out to demonstrate that this device meets the performance specifications for its intend use. The following tests were performed on the candidate device.

#### 7.1 Stability Summary:

The stability study was conducted to validate shelf life claim for the IMMULITE 2000 HCG Calibration Verification Material (CVM) to ensure that it maintains optimal product performance on IMMULITE 2000 platforms throughout the established shelf life of the CVM. The IMMULITE® 2000 HCG Calibration Verification Materials are stable up to 2.5 years when stored frozen at -20℃ prior to opening.

#### 7.1.1 Stability Protocol Summary:

The CVM study protocols are run as part of the calibrator stability testing. The stability CVMs and reference CVMs are run in duplicate (as a minimum) at the time points shown in Table 2 and the dose value determined from the reference calibrator curve.

| CVM Level | Time-Points (Days) | | | |
|-----------|--------------------|-----|-----|-----|
| LCGCVM1 | 1 | 548 | 730 | 912 |
| LCGCVM2 | 1 | 548 | 730 | 912 |
| LCGCVM3 | 1 | 548 | 730 | 912 |
| LCGCVM4 | 1 | 548 | 730 | 912 |

#### Table 2: Stability Time Points

#### 7.1.2 Stability Acceptance Criteria Summary:

The Acceptance Criteria for the IMMULITE HCG Calibration Verification Material (CVM) consists of 2 parts. Part 1 consists of the Guideline acceptance criteria which require dose value of stability calibrator/CVM to fall between ±12% of assigned dose for CVM level 2 and ±8% of assigned dose for CVM levels 3 and 4. Part 2 review limits criteria which require dose value of the controls to be within 2SD of the control target value when generated from the stability calibrator curve. If the result is not within acceptable dose range of ±12% for level 2 and ±8% for levels 3 and 4 then additional data review is conducted using part 2 criteria. The acceptance criterion is summarized in Table 3.

#### Table 3 Acceptance criteria for stability of IMMULITE 2000 HCG CVM

| CVM Level | Assigned Dose (mIU/mL) | Guideline Criteria % difference to assigned dose | Acceptable dose range (mIU/mL) | Review Limits |
|-----------|------------------------|--------------------------------------------------|--------------------------------|---------------|
| LCGCVM1 | 0.00 | N/A | <0.40 | Controls are |
| LCGCVM2 | 9.35 | $ \pm $ 12 | 8.23 - 10.47 | within 2SD |
| LCGCVM3 | 654 | $ \pm $ 8 | 601.68 - 706.32 | of target on |
| LCGCVM4 | 5327 | $ \pm $ 23 | 4101.79-6552.21 | each curve |

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#### 7.2 Traceability:

The IMMULITE HCG CVMs are traceable to WHO 3rd IS (75/537) and are manufactured using qualified materials and measurement procedures.

#### 7.3 Value Assignment:

HCG CVMs are 4 level materials which are a subset of 12 level HCG calibrators. Calibrators are not commercialized but are used internally during manufacture and release testing of HCG reagents and two point adjustors. The IMMULITE calibrators and therefore CVMs are value assigned using assigned reference calibrators. The assigned reference calibrators are prepared using HCG antigen spiked in a human serum matrix with preservatives and are traceable to WHO 3rd IS (75/537). Six levels of commercially available controls, and 30 pregnancy samples were used to validate calibrator/CVM value assignments.

The CVMs are manufactured using qualified materials and measurement procedures. The CVMs were tested on 15 replicates in total comprised of 5 runs and 3 replicates per run on 5 systems and 3 different reagent kit lots. The CVMs' dose values are generated using curve generated by assigned reference calibrators. The CVM values are calculated based on the recovered values for each run on each instrument independently. CVM values are then averaged across all systems. Quality control is performed by calculating the recovery of patient samples, spiked patient samples and controls using the assigned CVM values. The controls must fall within their target ranges.

#### 7.4 Expected Values/Reference Range:

Each CVM level was tested for a total of 15 replicates; 5 runs and 3 replicates per run. 3 different reagent kit lots and 5 different instruments were used to gain 15 replicates. The Guideline Range (95% confidence interval) for each CVM level was established based on the Target Mean and ± 2 Standard Deviation (SD). The expected values are provided in the IMMULITE® 2000 CVM Calibration verification Material lot-specific package insert. The expected assay range is 1 - 5000 mIU/mL. The target values in Table 4 can be considered as guidelines.

| Analyte<br>target levels | CVM Level | Target Mean<br>(mIU/mL) | Standard<br>Deviation<br>(SD) | Guideline ±2SD Range<br>(mIU/mL) | |
|--------------------------|-----------------|-------------------------|-------------------------------|----------------------------------|------|
| | CVM1 | 0.00 | - | 0.00 $\leq$ 0.40 | |
| | CVM2 | 5:4 | 0.405 | 4.59 | 6.21 |
| | CVM3 | 661 | 36.5 | 588 | 734 |
| | CVM4 | 4965 | 571 | 3823 | 6107 |
| Assay Range | 1 - 5000 mIU/mL | | | | |

#### Table 4: Target Values

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Each laboratory should establish their limits for acceptability based on methodology, clinical significance and medical decision levels of the test analyte. The representative, total precision tabulated in the respective assay instructions for use may be considered as one factor when establishing local, acceptable ranges. The values provided above may be considered as guidelines. Value assignment is lot specific.

Standard/Guidance Documents Referenced:

- CEN 13640 Stability Testing of In Vitro Diagnostic Reagents 0
- Guidance for Industry Abbreviated 510(k) Submissions for In Vitro Diagnostic . Calibrators
- Guidance for Industry and FDA Staff Assayed and Unassayed Quality Control o Material

#### Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10

#### 8. Conclusion:

The IMMULITE® 2000 HCG Calibration Verification Material is substantially equivalent to other products in commercial distribution intended for similar use. Most notably, it is substantially equivalent to the currently marketed Elecsys HCG CalCheck 5. The substantial equivalence of the device is supported by the non-clinical testing and was found to be comparable and supports the claims of substantial equivalence, product safety and effectiveness. Based on the testing completed and the comparisons with predicate device, the IMMULITE® 2000 HCG Calibration Verification Material does not raise any new questions on safety and effectiveness and the results support a determination of substantial equivalence.

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### 510(k) Summary

Introduction: According to the requirements of 21 CFR 807.92, the following information provides sufficient details to understand the basis for determination of substantial equivalence. The assigned 510(k) Number: k133128

| 1. Submitter | |
|--------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Mailing Address: | Siemens Healthcare Diagnostics Inc.<br>511 Benedict Avenue<br>Tarrytown, NY 10591 |
| Contact Person: | Asha Gartland |
| | Technical Regulatory Affairs Specialist |
| Phone Number: | (914)-524-3257 |
| Fax Number: | (914)-524-2101 |
| E-mail Address: | [e-mail] |
| Date Prepared: | October 16, 2013 |
| 2. Device Name | |
| Proprietary Name: | IMMULITE® 2000 Insulin Calibration Verification Material |
| Measurand: | Quality Control materials for IMMULITE® 2000 Insulin assay |
| | Calibration Verification Material (CVM) for IMMULITE® 2000 |
| Type of Test: | Insulin assay |
| Regulation Section: | 21 CFR 862.1660, Quality Control Material |
| Classification: | Class I Reserved |
| Products Code: | JJX - Single (Specified) Analyte Controls (Assayed and Unassayed) |
| Panel: | Clinical Chemistry (75) |
| 3. Predicate Device Name | Elecsys Insulin CalCheck 5 |
| Predicate 510(k) No: | K101075 |
| 4. Device Description: | The Calibration Verification Material (CVM) contains one set of four<br>vials, 2 mL each. CVMI contains an equine serum matrix with<br>preservatives. CVM2, CVM3 and CVM4 contain various levels of<br>insulin in an equine serum matrix with preservatives. |
| 5. Intended Use: | |
| Indication for Use: | See Indications for Use Statement below<br>The IMMULITE® Insulin Calibration Verification Material (CVM)<br>is for in vitro diagnostic use in the verification of calibration of the<br>IMMULITE Insulin assay on the IMMULITE 2000 systems |

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| Special Conditions for<br>Use Statement(s):<br>Special Instrument<br>Requirements: | For prescription use only<br>IMMULITE® 2000 Systems |
|--------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| 6. Technological<br>Characteristics and<br>Substantial Equivalence<br>Comparison with Predicate: | A comparison of the device features, intended use, and other<br>information demonstrates that the IMMULITE® 2000 Insulin<br>Calibration Verification Material (CVM) is substantially equivalent to<br>the predicate device as summarized in Table 1. |

### Table 1: Substantial Equivalence Comparison

| SIMILARITIES | | |
|--------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | Candidate Device | Predicate Device |
| | IMMULITE 2000 Insulin CVM | Elecsys Insulin CalCheck 5 |
| Intended Use | The IMMULITE® Insulin Calibration<br>Verification Material (CVM) is for in<br>vitro diagnostic use in the verification of<br>calibration of the IMMULITE Insulin<br>assay on the IMMULITE 2000 systems | The Elecsys Insulin CalCheck 5 is an<br>assayed control for use in calibration<br>verification and for use in the<br>verification of the assay range<br>established by the Elecsys Insulin<br>reagent on the indicated Elecsys and<br>cobas e immunoassay analyzers. |
| Analyte | Insulin | Same |
| Form | Lyophilized | Same |
| Stability | Stable unopened until the expiration date | Same |

| | DIFFERENCES | |
|---------|-----------------------------------------------|------------------------------------------------|
| | Candidate Device<br>IMMULITE 2000 Insulin CVM | Predicate Device<br>Elecsys Insulin CalCheck 5 |
| Matrix | Equine serum with preservatives | Bovine serum with preservatives |
| Storage | ≤20°C | 2-8 °C |
| Levels | 4 | 5 |
| Use | Single Use Only | Not For Single Use |

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#### 7. Non-Clinical Performance Testing

Performance testing has been carried out to demonstrate that this device meets the performance specifications for its intended use. The following tests were performed on the candidate device.

#### 7.1 Stability Summary:

The stability study was conducted to validate shelf life claim for the IMMULITE 2000 Insulin Calibration Verification Material (CVM) to ensure that it maintains optimal product performance on IMMULITE 2000 platforms throughout the established shelf life of the CVM. The IMMULITE® 2000 Calibration Verification Materials are stable up to 5 years when stored at -20℃ prior to opening.

#### 7.1.1 Stability Protocol Summary:

The CVM study protocols are run as part of the calibrator stability testing. The stability CVMs and reference CVMs are run in duplicate (as a minimum) at the time points shown in Table 2 and the dose value determined from the reference calibrator curve.

| CVM Level | Time-Points (Days) | | | |
|-----------|--------------------|------|------|------|
| LINCVMI | 1 | 1280 | 1460 | 1825 |
| LINCVM2 | 1 | 1280 | 1460 | 1825 |
| LINCVM3 | 1 | 1280 | 1460 | 1825 |
| LINCVM4 | 1 | 1280 | 1460 | 1825 |

#### Table 2: Stability Time Points

#### 7.1.2 Stability Acceptance Criteria Summary:

The Acceptance Criteria for the IMMULITE Insulin Calibration Verification Material (CVM) are in 2 parts. Part 1 consists of the guideline acceptance criteria which require dose value of stability calibrator/CVM to fall between ±12% of assigned dose for CVM level 2, ±10% of assigned dose for CVM levels 3 and ±13% for CVM level 4. Part 2 review limits criteria which require dose value of the controls to be within 2SD of the control target value when generated from the stability calibrator curve: If the result is not within acceptable dose range of ±12% for level 2, ±10% for levels 3 and ±13% for CVM level 4 then additional data review is conducted using part 2 criteria. The acceptance criterion is summarized in Table 3.

| CVM level | Assigned<br>Dose<br>( $\mu$ IU/mL) | Guideline Criteria<br>% difference to assigned<br>dose | Acceptable dose range<br>( $\mu$ IU/mL) | Review<br>Limits |
|-----------|------------------------------------|--------------------------------------------------------|-----------------------------------------|------------------|
| LINCVM1 | 0.00 | N/A | <2.00 | Controls are |
| LINCVM2 | 7.00 | $\pm$ 12% | 6.16 - 7.84 | within 2SD |
| LINCVM3 | 26.1 | $\pm$ 10% | 23.49 - 28.71 | of target on |
| LINCVM4 | 418 | $\pm$ 13% | 363.66 - 472.34 | each curve |

#### Table 3: Acceptance criteria for stability of IMMULITE 2000 Insulin CVM

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#### 7.2 Traceability:

The IMMULITE Insulin CVMs are traceable to WHO NIBSC 1st IRP (66/304). The CVMs are manufactured using qualified materials and measurement procedures.

#### 7.3 Value Assignment:

The Insulin CVMs are 4 level materials which are a subset of 10 level Insulin calibrators. Calibrators are not commercialized but are used internally during manufacture and release testing of Insulin reagents and two point adjustors. The IMMULITE calibrators and therefore CVMs are value assigned using assigned reference calibrators. The assigned reference calibrators are prepared using recombinant human Insulin antigen spiked into horse Serum with preservatives. Two levels of commercially available controls, and 30 patient samples (20 normal patients samples and 10 spiked samples) are used to validate calibrator/CVM value assignments.

The CVMs are manufactured using qualified materials and measurement procedures. The CVMs were tested on 26 replicates in total comprised of 9 runs and 2 or 3 replicates per run on 7 systems and 5 different reagent kit lots. The CVMs' dose values are generated using curve generated by assigned reference calibrators. The CVM values are calculated based on the recovered values for each run on each instrument independently. CVM values are then averaged across all systems. Quality control is performed by calculating the recovery of patient samples, spiked patient samples and controls using the assigned CVM values. The controls must fall within their target ranges.

#### 7.4 Expected Values/Reference Range:

Each CVM level was tested for a total of 26 replicates; 9 runs and 2 or 3 replicates per run. 5 different reagent kit lots and 7 different instruments were used to gain 26 replicates. The Guideline Range (95% confidence interval) for each CVM level was established based on the Target Mean and ± 2 Standard Deviation (SD). The expected values are provided in the IMMULITE® 2000 CVM Calibration Verification Material lot-specific package insert The expected assay range is 0.2 -300 µIU/mL. The target values in Table 4 can be considered as guidelines.

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| Analyte target<br>levels | CVM Level | Target Mean<br>(µIU/mL) | Standard<br>Deviation<br>(SD) | Guideline ±2SD<br>Range<br>(µIU/mL) | |
|--------------------------|-----------|----------------------------------|-------------------------------|-------------------------------------|------|
| | CVM1 | 0.00 | - | 0.00 | 2.00 |
| | CVM2 | 7.00 | 0.53 | 5.95 | 8.05 |
| | CVM3 | 26.1 | 1.3 | 23.5 | 28.7 |
| | CVM4 | 418 | - | - | - |
| | CVM4 | 314 *<br>(75% CVM4+<br>25% CVM1) | 20.5 | 273 | 355 |

| Assay Range | 0.2 -300 µIU/mL

* Note: CVM4 requires dilution to ensure the target value is within +10% of the top of the reportable range of the assay.

Each laboratory should establish their limits for acceptability based on methodology, clinical significance and medical decision levels of the test analyte. The representative, total precision tabulated in the respective assay instructions for use may be considered as one factor when establishing local, acceptable ranges. The values provided above may be considered as guidelines. Value assignment is lot specific.

#### Standard/Guidance Documents Referenced:

- CEN 13640 Stability Testing of In Vitro Diagnostic Reagents ●
- Guidance for Industry Abbreviated 510(k) Submissions for In Vitro Diagnostic . Calibrators
- o Guidance for Industry and FDA Staff - Assayed and Unassayed Quality Control Material

#### Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10

#### 8. Conclusion:

The IMMULITE® 2000 Insulin Calibration Verification Material is substantially equivalent to other products in commercial distribution intended for similar use. Most notably, it is substantially equivalent to the currently marketed Elecsys Insulin CalCheck 5. The substantial equivalence of the device is supported by the non-clinical testing and was found to be comparable and supports the claims of substantial equivalence, product safety and effectiveness. Based on the testing completed and the comparisons with predicate device, the IMMULITE® 2000 Insulin Calibration Verification Material does not raise any new questions on safety and effectiveness and the results support a determination of substantial equivalence.

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#### 510(k) Summary

Introduction: According to the requirements of 21 CFR 807.92, the following information provides sufficient details to understand the basis for determination of substantial equivalence. The assigned 510(k) Number: K133128

- 1. Submitter Mailing Address: Siemens Healthcare Diagnostics Inc. 511 Benedict Avenue Tarrytown, NY 10591 Contact Person: Asha Gartland Technical Regulatory Affairs Specialist Phone Number: (914)-524-3257 Fax Number: (914)-524-2101 E-mail Address: [e-mail] Date Prepared: October 16, 2013 IMMULITE® 2000 Pyrilinks-D Calibration Verification 2. Device Name Proprietary Name: Material Quality Control materials for IMMULITE® 2000 Pyrilinks-D Measurand: Type of Test: assav Calibration Verification Material (CVM) for IMMULITE® 2000 Pyrilinks-D assay Regulation Section: 21 CFR 862.1660, Quality Control Material Classification: Class I Reserved JJX – Single (Specified) Analyte Controls (Assayed and Products Code: Unassaved) Panel: Clinical Chemistry (75) IMMULITE® Unconjugated Estriol (uE3) Calibration 3. Predicate Device Name Verification Material (CVM) Predicate 510(k) No: K ! 1006 ! 4. Device Description: The Calibration Verification Material (CVM) contains one set of four vials, 2 mL each. CVM1 contains a phosphoric acid and sodium chloride matrix. CVM2, CVM3 and CVM4 contain various levels of H-Deoxypyridinoline in a phosphoric acid and sodium chloride matrix. 5. Intended Use: See Indications for Use Statement below Indication for Use: The IMMULITE® Pyrilinks-D Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE Pyrilinks-D assay on the IMMULITE 2000 systems

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| Special Conditions for Use Statement(s): | For prescription use only |
|------------------------------------------|---------------------------|
| Special Instrument Requirements: | IMMULITE® 2000 Systems |

6. Technological Characteristics and Substantial Equivalence Comparison with Predicate: A comparison of the device features, intended use, and other information demonstrates that the IMMULITE® 2000 Pyrilinks-D Calibration Verification Material (CVM) is substantially equivalent to the predicate device, as summarized in Table 1.

Table 1: Substantial Equivalence Comparison

| SIMILARITIES | | |
|-----------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | Candidate Device<br>IMMULITE 2000 Pyrilinks-D CVM | Predicate Device<br>IMMULITE uE3 Calibration<br>Verification Material (CVM) |
| Intended<br>Use | The IMMULITE® Pyrilinks-D<br>Calibration Verification Material<br>(CVM) is for in vitro diagnostic use in<br>the verification of calibration of the<br>IMMULITE Pyrilinks-D assay on the<br>IMMULITE 2000 systems. | For in vitro diagnostic use as a<br>control for the calibration<br>verification of the IMMULITE®<br>Unconjugated Estriol (uE3) assays<br>on the IMMULITE/IMMULITE<br>1000 and 2000 systems. |
| Form | Liquid | Same |
| Levels | 4 | Same |
| Stability | Stable unopened until the expiration<br>date | Same |
| Use | Single Use Only | Same |
| DIFFERENCES | | |
| | Candidate Device<br>IMMULITE 2000 Pyrilinks-D CVM | Predicate Device<br>IMMULITE uE3 Calibration<br>Verification Material (CVM) |
| Analyte | Pyrilinks-D | Unconjugated Estriol |
| Matrix | phosphoric acid and sodium chloride | Horse serum with preservatives |
| Storage | ≤20°C | 2-8°C |

### 7. Non-Clinical Performance Testing

Performance testing has been carried out to demonstrate that this device meets the performance specifications for its intended use. The following tests were performed on the candidate device.

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#### 7.1 Stability Summary:

The stability study was conducted to validate shelf life claim for the IMMULITE 2000 Pyrilinks-D Calibration Verification Material (CVM) to ensure that it maintains optimal product performance on IMMULITE 2000 platforms throughout the established shelf life of the CVM. The IMMULITE® 2000 Pyrilinks-D Calibration Verification Materials (CVMs) are stable up to5 years when stored at -20℃ prior to opening.

#### 7.1.1 Stability Protocol Summary:

The CVM study protocols are run as part of the calibrator stability testing. The stability CVMs and reference CVMs are run in duplicate (as a minimum) at the time points shown in Table 2 and the dose value determined from the reference calibrator curve.

| CVM Level | Time-Points (Days) | | | | |
|-----------|--------------------|------|------|------|------|
| LPDCVM1 | 1 | 1280 | 1460 | 1642 | 1825 |
| LPDCVM2 | 1 | 1280 | 1460 | 1642 | 1825 |
| LPDCVM3 | 1 | 1280 | 1460 | 1642 | 1825 |
| LPDCVM4 | 1 | 1280 | 1460 | 1642 | 1825 |

#### Table 2: Stability Time Points

7.1.2 Stability Acceptance Criteria Summary:

The Acceptance Criteria for the IMMULITE Pyrilinks-D Calibration Verification Material (CVM) are in 2 parts. Part 1 consists of the guideline acceptance criteria which require dose value of stability calibrator/CVM to fall between ±10% of assigned dose. Part 2 review limits criteria which require dose value of the controls to be within 2SD of the control target value when generated from the stability calibrator curve. If the result is not within acceptable dose range of ±10% then additional data review is conducted using part 2 criteria. The acceptance criterion is summarized in Table 3.

Table 3: Acceptance criteria for stability of IMMULITE 2000 Pyrilinks-D CVM

| CVM level | Assigned<br>Dose<br>(nmol/L) | Guideline Criteria<br>% difference to assigned<br>dose | Acceptable dose range<br>(nmol/L) | Review<br>Limits |
|-----------|------------------------------|--------------------------------------------------------|-----------------------------------|----------------------------------------------------------|
| LPDCVM1 | 0.00 | N/A | <7.00 | Controls are<br>within 2SD<br>of target on<br>each curve |
| LPDCVM2 | 18.30 | ±10% | 16.47 - 20.13 | |
| LPDCVM3 | 49.00 | ±10% | 44.10 - 53.90 | |
| LPDCVM4 | 302.00 | ±10% | 271.80 - 332.20 | |

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#### 7.2 Traceability:

The IMMULITE Pyrilinks-D CVMs are traceable to internal material which has been gravimetrically prepared. The CVMs are manufactured using qualified materials and measurement procedures.

#### 7.3 Value Assignment:

Pyrilinks-D CVMs are 4 level materials which are a subset of 8 level Pvrilinks-D calibrators. Calibrators are not commercialized but are used internally during manufacture and release testing of Pyrilinks-D reagents and two point adjustors. The IMMULITE calibrators and therefore CVMs are value assigned using assigned reference calibrators. The assigned reference calibrators are prepared using Hydrolysed Deoxypyridiniline antigen spiked in a matrix consisting of Phosphoric acid and Sodium Chloride in deionised water. Two levels of commercially available controls and 30 patient female urine samples are used to validate calibrator/CVM value assignments.

The CVMs are manufactured using qualified materials and measurement procedures. The CVMs were tested on 15 replicates in total comprised of 5 runs and 3 replicates per run on 4 systems and 3 different reagent kit lots. The CVMs' dose values are generated using curve generated by assigned reference calibrators. The CVM values are calculated based on the recovered values for each run on each instrument independently. CVM values are then averaged across all systems. Quality control is performed by calculating the recovery of patient samples, spiked patient samples and controls using the assigned CVM values. The controls must fall within their target ranges.

#### 7.4 Expected Values/Reference Range:

Each CVM level was tested for a total of 15 replicates; 5 runs and 3 replicates per run. 3 different reagent kit lots and 4 different instruments were used to gain 15 replicates. The Guideline Range (95% confidence interval) for each CVM level was established based on the Target Mean and ± 2 Standard Deviation (SD). The expected values are provided in the IMMULITE® 2000 CVM Calibration Verification Material lot-specific package insert. The expected assay range is 7 -300 nmol/L. The target values in Table 4 can be considered as guidelines.

| Analyte target<br>levels | CVM Level | Target Mean<br>(nmol/L) | Standard<br>Deviation<br>SD | Guideline ±2SD Range<br>(nmol/L) |
|--------------------------|----------------|-------------------------|-----------------------------|----------------------------------|
| | CVM1 | 0.00 | - | ≤7.00 |
| | CVM2 | 16.3 | 3.255 | 22.8 |
| | CVM3 | 49.2 | 5.4 | 60.0 |
| | CVM4 | 301 | 15 | 331 |
| Assay Range | 7 - 300 nmol/L | | | |

#### Table 4: Target Values

{15}------------------------------------------------

Each laboratory should establish their limits for acceptability based on methodology, clinical significance and medical decision levels of the test analyte. The representative, total precision tabulated in the respective assay instructions for use may be considered as one factor when establishing local, acceptable ranges. The values provided above may be considered as guidelines. Value assignment is lot specific.

Standard/Guidance Documents Referenced:

- CEN 13640 Stability Testing of In Vitro Diagnostic Reagents .
- Guidance for Industry Abbreviated 510(k) Submissions for In Vitro Diagnostic . Calibrators
- . Guidance for Industry and FDA Staff - Assayed and Unassayed Quality Control Material

#### Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10

#### Conclusion: 8.

The IMMULITE® 2000 Pyrilinks-D Calibration Verification Material is substantially equivalent to other products in commercial distribution intended for similar use. Most notably, it is substantially equivalent to the currently marketed IMMULITE Unconjugated Estriol (uE3) Calibration Verification Material (CVM). The substantial equivalence of the device is supported by the non-clinical testing and was found to be comparable and supports the claims of substantial equivalence, product safety and effectiveness. Based on the testing completed and the comparisons with predicate device, the IMMULITE® 2000 Pvrilinks-D Calibration Verification Material does not raise any new questions on safety and effectiveness and the results support a determination of substantial equivalence.

{16}------------------------------------------------

### 510(k) Summary

Introduction: According to the requirements of 21 CFR 807.92, the following information provides sufficient details to understand the basis for determination of substantial equivalence. The assigned 510(k) Number: K133128

| 1. Submitter<br>Mailing Address: | Siemens Healthcare Diagnostics Inc.<br>511 Benedict Avenue<br>Tarrytown, NY 10591 |
|--------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Contact Person:<br>Phone Number:<br>Fax Number:<br>E-mail Address:<br>Date Prepared: | Asha Gartland<br>Technical Regulatory Affairs Specialist<br>(914)-524-3257<br>(914)-524-2101<br>[e-mail]<br>October 16, 2013 |
| 2. Device Name<br>Proprietary Name:<br>Measurand:<br>Type of Test: | IMMULITE® 2000 Homocysteine Calibration Verification<br>Material<br>Quality Control materials for IMMULITE® 2000 Homocysteine assay<br>Calibration Verification Material (CVM) for IMMULITE® 2000<br>Homocysteine assay |
| Regulation Section:<br>Classification:<br>Products Code:<br>Panel: | 21 CFR 862.1660, Quality Control Material<br>Class I Reserved<br>JJX - Single (Specified) Analyte Controls (Assayed and Unassayed)<br>Clinical Chemistry (75) |
| 3. Predicate Device Name<br>Predicate 510(k) No: | Audit™ MicroCV™ Homocysteine Linearity Set<br>K100715 |
| 4. Device Description: | The Calibration Verification Material (CVM) contains one set of four<br>vials, 2mL each. CVM 1 contains a bovine protein/buffer matrix with<br>preservatives. CVM2, CVM3 and CVM4, contain various levels of s-<br>adenosyl-L-homocysteine in a bovine protein/buffer matrix with<br>preservatives. |
| 5. Intended Use:<br>Indication for Use: | See Indications for Use Statement below<br>The IMMULITE® Homocysteine Calibration Verification Materia<br>(CVM) is for in vitro diagnostic use in the verification of calibration o<br>the IMMULITE Homocysteine assay on the IMMULITE 2000<br>systems |
| Special Conditions for<br>Use Statement(s): | For prescription use only |
| Special Instrument | IMMULITE® 2000 Systems |
| Requirements: | |

{17}------------------------------------------------

- A comparison of the device features, intended use, and other 6. Technological Characteristics and Substantial Equivalence information demonstrates that the IMMULITE® 2000 Homocysteine Comparison with Predicate: Calibration Verification Material (CVM) is substantially equivalent to the predicate device, as summarized in Table 1.
Table 1: Substantial Equivalence Comparison

| SIMILARITIES | | |
|-----------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | Candidate Device<br>IMMULITE 2000 Homocysteine CVM | Predicate Device<br>Audit® MicroCVTM Homocysteine<br>Linearity |
| Intended<br>Use | The IMMULITE® Homocysteine<br>Calibration Verification Material (CVM) is<br>for in vitro diagnostic use in the<br>verification of calibration of the<br>IMMULITE Homocysteine assay on the<br>IMMULITE 2000 systems. | The Audit® MicroCVTM Homocysteine<br>Linearity for use with Siemens<br>Centaur® Immunoassay<br>Systems, is intended to simulate human<br>patient serum samples for purpose of<br>determining linearity, calibration<br>verification and verification of<br>reportable range for Homocysteine. |
| Analyte | Homocysteine | Same |
| Stability | Stable unopened until the expiration date | Same |
| Form | Liquid | Same |

| DIFFERENCES | | |
|-------------|----------------------------------------------------|----------------------------------------------------------------|
| | Candidate Device<br>IMMULITE 2000 Homocysteine CVM | Predicate Device<br>Audit® MicroCVTM Homocysteine<br>Linearity |
| Matrix | Bovine protein/buffer matrix | Human serum |
| levels | 4 | 5 |
| Storage | ≤20°C | 2-8°C |
| Use | Single Use Only | Not For Single Use |

{18}------------------------------------------------

### 7. Non-Clinical Performance Testing

Performance testing has been carried out to demonstrate that this device meets the performance specifications for its intend use. The following tests were performed on the candidate device.

#### 7.1 Stability Summary:

The stability study was conducted to validate shelf life claim for the IMMULITE 2000 Homocysteine Calibration Verification Material (CVM) to ensure that it maintains optimal product performance on IMMULITE 2000 platforms throughout the established shelf life of the CVM. The IMMULITE® 2000 Homocysteine Calibration Verification Materials (CVMs) are stable up to 3.5 years when stored at -20°C prior to opening.

7.1.1 Stability Protocol Summary:

The CVM study protocols are run as part of the calibrator stability testing. The stability CVMs and reference CVMs are run in duplicate (as a minimum) at the time points shown in Table 2 and the dose value determined from the reference calibrator curve.

| CVM Level | Time-Points (Days) | | | | |
|-----------|--------------------|-----|-----|--------|------|
| LHOCVM1 | | 730 | 912 | । 095 | 1280 |
| LHOCVM2 | | 730 | 912 | । 0652 | 1280 |
| LHOCVM3 | | 730 | 912 | । 095 | 1280 |
| LHOCVM4 | | 730 | 912 | । 092 | 1280 |

#### Table 2: Stability Protocol Summary

Stability Acceptance Criteria Summary:

The Acceptance Criteria for the IMMULITE Homocysteine Calibration Verification Material (CVM) are in 2 parts. Part 1 consists of the guideline acceptance criteria which require dose value of stability calibrator/CVM to fall between ±15% of assigned dose for CVM level 2, ±10% of assigned dose for CVM levels 3 and 4. Part 2 review limits criteria which require dose value of the controls to be within 2SD of the control target value when generated from the stability calibrator curve. If the result is not within acceptable dose range of ±15% for level 2 and ±10% for levels 3 and 4 then additional data review is conducted using part 2 criteria. The acceptance criterion is summarized in Table 3.

| CVM level | Assigned<br>Dose<br>(µmol/L) | Guideline Criteria<br>% difference to assigned<br>dose | Acceptable dose range<br>(µmol/L) | Review<br>Limits |
|-----------|------------------------------|--------------------------------------------------------|-----------------------------------|----------------------------------------------------------|
| LHOCVM1 | 0.00 | N/A | <2.0 | Controls are<br>within 2SD<br>of target on<br>each curve |
| LHOCVM2 | 4.12 | ±15% | 3.50 - 4.74 | |
| LHOCVM3 | 17.60 | ±10% | 15.84 - 19.36 | |
| LHOCVM4 | 71.50 | ±10% | 64.35 - 78.65 | |

#### Table 3 : Acceptance criteria for stability of IMMULITE 2000 Homocysteine CVM

{19}------------------------------------------------

#### 7.2 Traceability:

The IMMULITE Homocysteine CVMs are traceable to internal material which has been gravimetrically prepared. The CVMs are manufactured using qualified materials and measurement procedures.

#### 7.3 Value Assignment:

Homocysteine CVMs are 4 level materials which are a subset of 7 level Homocysteine calibrators. Calibrators are not commercialized but are used internally during manufacture and release testing of Homocysteine reagents and two point adjustors. The IMMULITE calibrators and therefore CVMs are value assigned using assigned reference calibrators. The assigned reference calibrators are prepared using s-adenosyl-L-homocysteine spiked into a Bovine protein/buffer matrix and are traceable to internal material which has been gravimetrically prepared. Two levels of commercially available controls and 40 samples (30 spiked samples, 5 normal samples and 5 patient samples) are used to validate calibrator/CVM value assignments.

The CVMs are manufactured using qualified materials and measurement procedures. The CVMs were tested on 27 replicates in total comprised of 9 runs and 3 replicates per run on 7 systems and 3 different reagent kit lots. The CVMs' dose values are generated using curve generated by assigned reference calibrators. The CVM values are calculated based on the recovered values for each run on each instrument independently. CVM values are then averaged across all systems. Quality control is performed by calculating the recovery of patient samples, spiked patient samples and controls using the assigned CVM values. The controls must fall within their target ranges.

#### 7.4 Expected Values/Reference Range:

Each CVM level was tested for a total of 27 replicates; 9 runs and 3 replicates per run, 3 different reagent kit lots and 7 different instruments were used to gain 27 replicates. The Guideline Range (95% confidence interval) for each CVM level was established based on the Target Mean and ± 2 Standard Deviation (SD). The expected values are provided in the IMMULITE® 2000 CVM Calibration verification Material lot-specific package insert. The expected assay range is 2 -50 umol/L. The target values in Table 4 can be considered as guidelines.

{20}------------------------------------------------

| Analyte target<br>levels | CVM Levels | Target Mean<br>(µmol/L) | Standard<br>Deviation<br>(SD) | Guideline ±2SD<br>Range<br>(µmol/L) | |
|--------------------------|-------------|-----------------------------------|-------------------------------|-------------------------------------|------|
| | CVM1 | 0.00 | - | 0.00 | ≤2.0 |
| | CVM2 | 4.11 | 0.43 | 3.25 | 4.97 |
| | CVM3 | 17.5 | 0.875 | 15.8 | 19.3 |
| | CVM4 | 73.0 | - | - | - |
| | | 51.1*<br>(70% CVM4 +<br>30% CVM1) | 2.55 | 46.0 | 56.2 |
| Assay Range | 2-50 µmol/L | | | | |

Table 4: Target Values

**Assay Range** 2 -50 μmol/L

* Note: CVM4 requires dilution to ensure the target value is within +10% of the top of the reportable range of the assay.

Each laboratory should establish their limits for acceptability based on methodology, clinical significance and medical decision levels of the test analyte. The representative, total precision tabulated in the respective assay instructions for use may be considered as one factor when establishing local, acceptable ranges. The values provided above may be considered as guidelines. Value assignment is lot specific.

#### Standard/Guidance Documents Referenced:

- CEN 13640 Stability Testing of In Vitro Diagnostic Reagents ●
- Guidance for Industry Abbreviated 510(k) Submissions for In Vitro Diagnostic . Calibrators
- Guidance for Industry and FDA Staff Assayed and Unassayed Ouality Control . Material

#### Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10

#### 8. Conclusion:

The IMMULITE® 2000 Homocysteine Calibration Verification Material (CVM) is substantially equivalent to other products in commercial distribution intended for similar use. Most notably, it is substantially equivalent to the currently marketed Audit® MicroCVTM Homocysteine Linearity for use with Siemens Centaur® Immunoassay Systems. The substantial equivalence of the device is supported by the non-clinical testing and was found to be comparable and supports the claims of substantial equivalence, product safety and effectiveness.

{21}------------------------------------------------

Based on the testing completed and the comparisons with predicate device, the IMMULITE® 2000 Homocysteine Calibration Verification Material does not raise any new questions on safety and effectiveness and the results support a determination of substantial equivalence.

{22}------------------------------------------------

#### 510(k) Summary

Introduction: According to the requirements of 21 CFR 807.92, the following information provides sufficient details to understand the basis for determination of substantial equivalence.

#### The assigned 510(k) Number: k133128

- 1. Submitter Mailing Address:
Contact Person:

Phone Number: Fax Number: E-mail Address: Date Prepared:

- 2. Device Name Proprietary Name: Measurand:
Type of Test:

Regulation Section: Classification: Products Code: Panel:

3. Predicate Device Name

Predicate 510(k) No:

4. Device Description:

- 5. Intended Use: Indication for Use:
Siemens Healthcare Diagnostics Inc. 511 Benedict Avenue Tarrytown, NY 10591

Asha Gartland Technical Regulatory Affairs Specialist (914)-524-3257 (914)-524-2101 [e-mail] October 16, 2013

#### IMMULITE® 2000 Growth Hormone Calibration Verification Material

Quality Control materials for IMMULITE® 2000 Growth Hormone assav Calibration Verification Material (CVM) for IMMULITE® 2000 Growth Hormone assay

21 CFR 862.1660, Quality Control Material Class I Reserved

JJX - Single (Specified) Analyte Controls (Assayed and Unassayed) Clinical Chemistry (75)

IMMULITE® Unconjugated Estriol (uE3) Calibration Verification Material (CVM) K110061

The Calibration Verification Material (CVM) contains one set of four vials, 2 mL each. CVM 1 contains an equine serum matrix with preservatives. CVM2, CVM3 and CVM4 contain various levels of human growth hormone in equine serum matrix with preservatives.

See Indications for Use Statement below The IMMULITE® Growth Hormone Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE Growth Hormone assay on the IMMULITE 2000 systems.

{23}------------------------------------------------

Special Conditions for Use Statement(s): Special Instrument Requirements:

- 6. Technological Characteristics and Substantial Equivalence Comparison with Predicate:
For prescription use only

IMMULITE® 2000 Systems

A comparison of the device features, intended use, and other information demonstrates that the IMMULITE® 2000 Growth Hormone Calibration Verification Material (CVM) is substantially equivalent to the predicate device, as summarized in Table 1.

#### Table 1: Substantial Equivalence Comparison

| SIMILARITIES | | |
|-----------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | Candidate Device<br>IMMULITE 2000 Growth Hormone CVM | Predicate Device<br>IMMULITE uE3Calibration<br>Verification Material (CVM) |
| Intended<br>Use | The IMMULITE® Growth Hormone<br>Calibration Verification Material (CVM) is<br>for in vitro diagnostic use in the verification<br>of calibration of the IMMULITE Growth<br>Hormone assay on the IMMULITE 2000<br>systems. | For in vitro diagnostic use as a<br>control for the calibration verification<br>of the IMMULITE® Unconjugated<br>Estriol (uE3) assays on the<br>IMMULITE/IMMULITE 1000 and<br>2000 systems. |
| Matrix | Equine serum with preservatives | Same |
| Levels | 4 | Same |
| Stability | Stable unopened until the expiration<br>date on the vial | Same |
| Storage | 2-8°C | Same |
| Use | Single Use Only | Same |

| DIFFERENCES | | |
|-------------|------------------------------------------------------|-----------------------------------------------------------------------------|
| | Candidate Device<br>IMMULITE 2000 Growth Hormone CVM | Predicate Device<br>IMMULITE uE3 Calibration<br>Verification Material (CVM) |
| Analyte | Growth Hormone | Unconjugated Estriol |
| Form | Lyophilized | Liquid |

{24}------------------------------------------------

#### 7. Non-Clinical Performance Testing

Performance testing has been carried out to demonstrate that this device meets the performance specifications for its intend use. The following tests were performed on the candidate device.

#### 7.1 Stability Summary:

The stability study was conducted to validate shelf life claim for the IMMULITE 2000 Growth Hormone Calibration Verification Material (CVM) to ensure that it maintains optimal product performance on IMMULITE 2000 platforms throughout the established shelf life of the CVM. The IMMULITE® 2000 Growth Hormone Calibration Verification Materials (CVMs) are stable up to 6 years when stored at 2-8°C prior to opening.

#### 7.1.1 Stability Protocol Summary:

The CVM study protocols are run as part of the calibrator stability testing. The stability CVMs and reference CVMs are run in duplicate (as a minimum) at the time points shown in Table 2 and the dose value determined from the reference calibrator curve.

| CVM Level | Time-Points (Days) | | | |
|-----------|--------------------|------|------|------|
| LGRHCVM1 | 1 | 1280 | 1642 | 2190 |
| LGRHCVM2 | 1 | 1280 | 1642 | 2190 |
| LGRHCVM3 | 1 | 1280 | 1642 | 2190 |
| LGRHCVM4 | 1 | 1280 | 1642 | 2190 |

#### Table 2: Stability Protocol Summary

#### 7.1.2 Stability Acceptance Criteria Summary:

The Acceptance Criteria for the IMMULITE Growth Hormone Calibration Verification Material (CVM) are in 2 parts. Part 1 consists of the guideline acceptance criteria which require dose value of stability calibrator/CVM to fall between ±12% of assigned dose for CVM levels 2 and 4, ±10% of assigned dose for CVM level 3. Part 2 review limits criteria which require dose value of the controls to be within 2SD of the control target value when generated from the stability calibrator curve. If the result is not within acceptable dose range of ±12% for levels 2 and 4, ±10% for levels 3 then additional data review is conducted using part 2 of the criteria. The acceptance criterion is summarized in Table 3.

{25}------------------------------------------------

| CVM level | Assigned<br>Dose<br>(ng/mL) | Guideline Criteria<br>% difference to assigned<br>dose | Acceptable dose range<br>(ng/mL) | Review<br>Limits |
|-----------|-----------------------------|--------------------------------------------------------|----------------------------------|------------------|
| LGRHCVMI | 0.00 | N/A | <0.01 | Controls are |
| LGRHCVM2 | 0.61 | ±12% | 0.54 - 0.68 | within 2SD |
| LGRHCVM3 | 9.6 | ±10% | 8.64 - 10.56 | of target on |
| LGRHCVM4 | 40 | ±12% | 35.20 - 44.80 | each curve |

Table 3: Acceptance criteria for stability of IMMULITE 2000 Growth Hormone CVM

#### 7.2 Traceability:

The IMMULITE Growth Hormone CVMs are trace…