SEALITE SCIENCES, INC. AQUALITE FERRITIN ASSAY

K970612 · Sealite Sciences, Inc. · DBF · Jun 24, 1997 · Immunology

Device Facts

• Record ID: K970612
• Device Name: SEALITE SCIENCES, INC. AQUALITE FERRITIN ASSAY
• Applicant: Sealite Sciences, Inc.
• Product Code: DBF · Immunology
• Decision Date: Jun 24, 1997
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5340
• Device Class: Class 2

Intended Use

The AquaLite® Ferritin Bioluminescent Immunoassay (BIA) Kit (or AquaLite® Ferritin Assay) is an in vitro diagnostic product intended for use in clinical laboratories for the quantitative measurement of ferritin in human serum and plasma. Ferritin measurements are used in the diagnosis of diseases affecting iron metabolism.

Device Story

The AquaLite® Ferritin Assay is a bioluminescent immunoassay (BIA) for quantitative ferritin measurement in human serum or plasma. The device uses mouse monoclonal anti-ferritin antibodies pre-coated on polystyrene tubes and a secondary antibody conjugated to AquaLite®. Samples are pipetted into tubes; ferritin forms a sandwich complex with the antibodies. After incubation and washing, a calcium trigger buffer is injected, causing the AquaLite® conjugate to oxidize its self-contained luciferin, emitting a flash of light. A luminometer measures the light intensity (RLU), which is directly proportional to ferritin concentration. Results are calculated via a calibration curve plotting RLU against known ferritin concentrations. The assay is performed in clinical laboratories by trained personnel. It aids clinicians in diagnosing iron metabolism disorders, including iron overload and iron deficiency anemia. High-dose samples require dilution. The device provides a diagnostic tool for assessing iron status.

Clinical Evidence

Bench testing only. Performance characteristics established: sensitivity (0.03 ng/mL), intra-assay precision (3.6-8.6% CV), inter-assay precision (3.7-9.3% CV), and linearity. Method comparison with a commercial chemiluminometric immunoassay (N=99) showed a correlation coefficient of 0.98. Interference testing confirmed no significant impact from hemoglobin (500 mg/dL), bilirubin (20 mg/dL), or triglycerides (3000 mg/dL). No high-dose hook effect observed below 10,000 ng/mL.

Technological Characteristics

Bioluminescent immunoassay (BIA) using mouse monoclonal anti-ferritin antibodies. Solid phase: polystyrene tubes. Detection: flash-type bioluminescence triggered by calcium buffer oxidation of self-contained luciferin. Measured via luminometer. Sample volume: 25 µL. Incubation: 60 minutes at room temperature. Preservative: sodium azide.

Indications for Use

Indicated for the quantitative measurement of ferritin in human serum and plasma in clinical laboratory settings to aid in the diagnosis of diseases affecting iron metabolism, such as hemochromatosis and iron deficiency anemia.

Regulatory Classification

Identification

A ferritin immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the ferritin (an iron-storing protein) in serum and other body fluids. Measurements of ferritin aid in the diagnosis of diseases affecting iron metabolism, such as hemochromatosis (iron overload) and iron deficiency amemia.

Predicate Devices

• Chiron Diagnostics ACS:180 Ferritin Assay

Submission Summary (Full Text)

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K970612

SeaLite Sciences, Inc.

JUN 24 1997

# 510(k) SUMMARY

## I. GENERAL INFORMATION

Trade or proprietary name - SeaLite Sciences, Inc. AquaLite® Ferritin Assay

Common or usual name - Bioluminescent immunoassay (BIA)

Classification name - FDA has classified ferritin test systems intended for the measurement of ferritin in the diagnosis of diseases affecting iron metabolism, such as hemochromatosis (iron overload) and iron deficiency anemia as Class II devices. (21 C.F.R. § 866.5340)

Submitter’s Name and Address:
Cathryn C. Cambria
Director, Regulatory Affairs and Quality Assurance
SeaLite Sciences, Inc.
3000 Northwoods Parkway
Suite 200
Norcross, GA 30071
[phone], ext. 227

Submission Date:
February 14, 1997

Legally Marketed Device To Which Claim Substantial Equivalence:
Chiron Diagnostics ACS:180 Ferritin Assay

## II. INDICATIONS FOR USE

The AquaLite® Ferritin Bioluminescent Immunoassay (BIA) Kit (or the AquaLite® Ferritin Assay) is an *in vitro* diagnostic product intended for use in clinical laboratories for the quantitative determination of human ferritin in serum and plasma. Ferritin measurements are used in the diagnosis of diseases affecting iron metabolism.

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## III. DEVICE DESCRIPTION

The AquaLite® Ferritin Bioluminescent Immunoassay Kit uses a mouse monoclonal anti-ferritin antibody that is pre-coated onto polystyrene tubes (solid phase). Samples (serum or plasma), or appropriate calibrators or controls, are pipetted (25 µL) into the pre-coated tubes. A mouse monoclonal anti-ferritin antibody covalently linked to AquaLite® (150 µL) is then added to the tubes. Ferritin in the sample simultaneously combines with the antibody on the solid phase and conjugate antibody to form an immune complex or "sandwich" bound to the solid phase. Complex formation is complete after a 60-minute incubation period at room temperature on a standard orbital shaker. The tubes are then washed to remove unbound conjugate.

The washed tubes are placed in a luminometer that is capable of reading a triggered, flash-type reaction in 12 x 75 mm tubes. Injection of the calcium trigger buffer causes AquaLite® to oxide its self-contained luciferin molecule, producing a flash of light, which is measured by the luminometer. The intensity of the light emitted from antibody bound to the tubes is directly proportional to the concentration of the ferritin in the sample. To calculate results, the light intensity (in relative light units, RLU) of the ferritin calibrators is plotted against ferritin concentration (in ng per mL) to yield a calibration curve. This curve is used to relate the light intensity generated from the samples and controls to ferritin concentration in ng/mL.

Note: Samples that generate signals greater than the signal from the highest calibrator are off-scale. These samples must be diluted and re-assayed. Remember to multiply the results from diluted samples by the dilution factor used.

## IV. SUMMARY OF STUDIES AND TECHNOLOGICAL CHARACTERISTICS

Studies on the AquaLite® Ferritin Assay were conducted at SeaLite Sciences. The results are summarized below:

### Performance Characteristics

#### 1. Sensitivity

The sensitivity or detection limit of the AquaLite® Ferritin Assay is 0.03 ng/mL. Sensitivity is determined by adding the mean signal of 20 replicates of the zero level calibrator plus two (2) standard deviations above this mean. The ferritin concentration (ng/mL) associated with this calculated signal is defined as the sensitivity of the assay.

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## 2. Specificity

The AquaLite® Ferritin Assay measures intact ferritin. Cross reactivity of the AquaLite® Ferritin assay was determined by measuring the concentration of ferritin in 2 human serum samples spiked with 300 ng/mL liver ferritin. Percent cross-reactivity (%) is reported below:

| Endogenous Ferritin (ng/mL) | Ferritin Expected (ng/mL) | Ferritin Observed (ng/mL) | Cross-Reactivity (%) |
| --- | --- | --- | --- |
| 79 | 379 | 323 | 85 |
| 139 | 439 | 451 | 103 |

## 3. High Dose Hook Effect

No high dose hook effect occurs prior to 10,000 ng/mL ferritin.

## 4. Precision

(a) Intra-assay precision. Two serum commercial controls containing ferritin at the following concentrations were assayed to determine intra-assay precision. (Total N = 20 per concentration level.)

| Ferritin Level (ng/mL) | % CV |
| --- | --- |
| 368.4 | 8.6 |
| 123.9 | 3.6 |
| 40.5 | 5.4 |

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(b) Inter-assay precision. Commercial controls containing ferritin at the following concentrations were assayed in duplicate repetitively. Ten (10) assays were performed and a new standard curve was generated for each assay. The inter-assay precision observed for the solutions (Total n = 2 x 10 = 20) are shown below.

| Ferritin Level (ng/mL) | % CV |
| --- | --- |
| 50.9 | 4.5% |
| 121.6 | 3.7% |
| 363.2 | 9.3% |

## 5. Method Comparison

The AquaLite® Ferritin Assay was used to assay patient samples (N=99) that were previously assayed by a commercially available chemiluminometric immunoassay. A slope of 0.72 with a y-intercept of 15.9 was obtained. The correlation coefficient was 0.98.

## 6. Linearity and Nonparallelism

Three human serum samples containing the levels of endogenous ferritin shown below were diluted as indicated using Calibrator A (0 ng/mL) and assayed in duplicate. All concentrations are in ng/mL.

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| SAMPLE ID | DILUTION FACTOR | OBSERVED (ng/mL) | FERRITIN EXPECTED (ng/mL) | RECOVERY (%) |
| --- | --- | --- | --- | --- |
| A | Undiluted | 160.1 | --- | --- |
| | 1:2 | 77.3 | 80 | 96.6 |
| | 1:4 | 47.4 | 40 | 118.5 |
| | 1:8 | 23.9 | 20 | 119.5 |
| B | Undiluted | 294.1 | --- | --- |
| | 1:2 | 160.9 | 147 | 109.5 |
| | 1:4 | 84.4 | 74 | 114.1 |
| | 1:8 | 45.1 | 27 | 121.9 |
| C | Undiluted | 878 | --- | --- |
| | 1:2 | 383.0 | 439 | 87.2 |
| | 1:4 | 189.6 | 219.5 | 86.4 |
| | 1:8 | 101.4 | 109.8 | 92.3 |

## 7. Spike and Recovery

Three normal human serum samples were diluted with 120 ng/mL human spleen ferritin (WHO 2nd IS 80/578). The spiked samples were assayed using the AquaLite® Ferritin Assay. All values are in ng/mL.

| Sample | Unspiked (ng/mL) | Spike Recovered (ng/mL) | Spiked Expected (ng/mL) | % |
| --- | --- | --- | --- | --- |
| 1 | 1260 | 595 | 690 | 86 |
| 2 | 58 | 90 | 90 | 100 |
| 3 | 406 | 221 | 263 | 84 |

## 8. Recovery in Serum and Plasma

Blood samples from 6 normal subjects were prepared as sera (standard tubes) as well as EDTA plasma. Ferritin was quantified using the AquaLite® Ferritin Assay. Recovered ferritin was compared with the ferritin recovered in serum (standard technique). The data demonstrate that there are no significant differences among serum and EDTA plasma when using the AquaLite® Ferritin Assay.

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| Sample | Serum (ng/mL) | EDTA (ng/ml) | % |
| --- | --- | --- | --- |
| 1 | 63.9 | 62.0 | 97 |
| 2 | 44.1 | 38.7 | 88 |
| 3 | 62.8 | 53.6 | 85 |
| 4 | 109.6 | 110.9 | 101 |
| 5 | 105.2 | 108.8 | 103 |
| 6 | 63.3 | 62.6 | 99 |

## 9. Effect of Common Interferents

Pooled normal human serum was spiked with preparations of hemoglobin, bilirubin, and triglycerides to the levels shown below. Ferritin was quantified using the AquaLite® Ferritin assay. Recovered ferritin was compared to the ferritin recovered in normal serum. The data demonstrate that the AquaLite® Ferritin assay is not significantly affected by hemoglobin, bilirubin, or triglycerides at the levels tested.

| Analyte | Observed | Expected | % |
| --- | --- | --- | --- |
| Hgb at 500 mg/dL | 113.3 | 115.6 | 98 |
| Bilirbn at 20 mg/dL | 114.2 | 115.6 | 90 |
| Trig at 3000 mg/dL | 116.2 | 128.9 | 99 |

## V. POTENTIAL ADVERSE EFFECTS OF THE DEVICE ON HEALTH

Caution: Use Universal Precautions. No known test method can offer complete assurance that products derived from human serum are pathogen-free; therefore, handle all materials of human origin as though they were potentially infectious.

Sodium azide is used as a preservative. This preservative may react with metallic plumbing to from explosive metal azides. Flush with large volumes of water when disposing of materials containing sodium azide.

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As an *in vitro* diagnostic test, there are not direct adverse effects on the health of a patient from the use of this product. However, failure of the device to perform as indicated, the contamination of reagents, the use of reagents past the labeled expiration dates, the use of improper specimens, or human error during the performance of the test may lead to erroneous results and possible improper patient management.

## VI. CONCLUSIONS DRAWN FROM STUDIES

The data from the studies conducted demonstrate that the performance of SeaLite Sciences, Inc. AquaLite® Ferritin Assay is similar and substantially equivalent to that of other commercially available assays for ferritin.

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DEPARTMENT OF HEALTH & HUMAN SERVICES
Public Health Service
Food and Drug Administration
2098 Gaither Road
Rockville MD 20850

Cathryn C. Cambria
Director, Regulatory Affairs & Quality Assurance
SeaLite Sciences, Inc.
3000 Northwoods Parkway
Suite 200
Norcross, GA 30071

JUN 24 1997

Re: K970612/S001
Trade Name: SeaLite Sciences, Inc. AquaLite® Ferritin Assay
Regulatory Class: II
Product Code: DBF
Dated: May 08, 1997
Received: May 12, 1997

Dear Ms. Cambria:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Good Manufacturing Practice for Medical Devices: General (GMP) regulation (21 CFR Part 820) and that, through periodic GMP inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal Laws or Regulations.

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at [phone].

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This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for *in vitro* diagnostic devices), please contact the Office of Compliance at [phone]. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll free number [phone] or at [phone] or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours,

Steven I. Gutman, M.D., M.B.A.
Director
Division of Clinical Laboratory Devices
Office of Device Evaluation
Center for Devices and Radiological Health

Enclosure

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Page 1 of 1

510(k) Number (if known): ___________

Device Name: AquaLite® Ferritin Bioluminescent Immunoassay (BIA) Kit (or the AquaLite® Ferritin Assay)

Indications for Use:

The AquaLite® Ferritin Bioluminescent Immunoassay (BIA) Kit (or AquaLite® Ferritin Assay) is an *in vitro* diagnostic product intended for use in clinical laboratories for the quantitative measurement of ferritin in human serum and plasma. Ferritin measurements are used in the diagnosis of diseases affecting iron metabolism.


(Division Sign-Off)
Division of Clinical Laboratory Devices
510(k) Number ___________

(PLEASE DO NOT WRITE BELOW THIS LINE CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Prescription Use ☑ (Per 21 CFR 801.109)
OR
Over-The-Counter Use ☐ (Optional Format 1-2-96)

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