Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a standard immunoassay system for quantitative determination of a biomarker (HE4). There is no mention of AI, ML, image processing, or any other technology typically associated with AI/ML in the device description, intended use, or performance studies. The performance metrics are standard for a diagnostic assay, not indicative of an AI/ML algorithm's performance.

Therapeutic

No

This device is for in vitro diagnostic use, specifically for quantitative determination of HE4 in human samples as an aid in monitoring recurrence or progressive disease, not for direct therapy.

Diagnostic

Yes
The document states that the device is "For in vitro diagnostic use" and is "to be used as an aid in monitoring recurrence or progressive disease in patients with epithelial ovarian cancer," which directly indicates its diagnostic purpose.

SaMD (Software as a Medical Device)

No

The device description explicitly states it is an "assay system, including a set of immunoassay reagents," which are physical components, not software. It also mentions operation on the "LUMIPULSE G System," which is a hardware platform.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Explicit Statement: The "Intended Use / Indications for Use" section clearly states: "For in vitro diagnostic use."
Nature of the Device: The device is an "assay system, including a set of immunoassay reagents, for the quantitative measurement of HE4 in specimens." This describes a test performed on biological samples (serum and plasma) outside of the body, which is the definition of an in vitro diagnostic.
Intended Use: The intended use is to "aid in monitoring recurrence or progressive disease in patients with epithelial ovarian cancer" by measuring a specific analyte (HE4) in patient samples. This is a diagnostic purpose.
Calibrators: The inclusion of calibrators for use in the "quantitative measurement of HE4" further supports its role as a diagnostic test.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Lumipulse G HE4 Immunoreaction Cartridges:

For in vitro diagnostic use.

Lumipulse G HE4 is a Chemiluminescent Enzyme Immunoassay (CLEIA) for the quantitative determination of HE4 in human serum and plasma (lithium heparin or dipotassium EDTA) on the Lumipulse G System.

The assay is to be used as an aid in monitoring recurrence or progressive disease in patients with epithelial ovarian cancer. Serial testing for patient HE4 values should be used in conjunction with other clinical methods used for monitoring epithelial ovarian cancer.

Lumipulse G HE4 Calibrators:

Lumipulse G HE4 Calibrators are for use in the calibration of the Lumipulse G System for the quantitative measurement of HE4 in human serum and plasma (lithium heparin or dipotassium EDTA).

Product codes (comma separated list FDA assigned to the subject device)

OIU, JIT

Device Description

The Lumipulse G HE4 is an assay system, including a set of immunoassay reagents, for the quantitative measurement of HE4 in specimens based on CLEIA technology by a two-step sandwich immunoassay method on the LUMIPULSE G System.

Lumipulse G HE4 Immunoreaction Cartridges |REF| 234174

The Lumipulse G HE4 Immunoreaction Cartridges consists of 3 × 14 tests. Each kit contains the following:

Antibody-Coated Particle Solution (Liquid when used, 250 µL/Immunoreaction Cartridge)

Contains 150 ug/mL anti-HE4 monoclonal antibody (mouse)-coated particles, protein stabilizers (bovine and mouse) and chemical stabilizers in 0.15 M sodium chloride/Tris buffer. This solution contains gelatin and turns into gel at 15 °C or lower. Preservative: Proclin 300.

Enzyme-Labeled Antibody Solution
(Liquid, 350 µL/Immunoreaction Cartridge)

Contains 0.25 µg/mL alkaline phosphatase (ALP: calf) labeled anti-HE4 monoclonal antibody (mouse), protein stabilizers (bovine, calf and mouse) and chemical stabilizers in 0.1 M sodium chloride/MES buffer. Preservative: Proclin 300.

LUMIPULSE G HE4 Calibrators CAL 234181

Each calibrator kit contains one bottle each of Calibrators 1 and 2. The calibrator kit is packaged separately.

CAL Liquid, 1 × 2 Concentrations

CAL 1 0 pM HE4 calibrator (1 × 1.5 mL)

CAL 2 1500 pM HE4 calibrator (1 × 1.5 mL)

Contains sodium chloride and potassium chloride in phosphate buffer with protein stabilizer (bovine). Preservative: Proclin 950.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Clinical and Hospital laboratories

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Precision/Reproducibility:

Lumipulse G HE4 demonstrated precision = 0.9.

Clinical Supportive Data - Monitoring of Disease status in Patients Diagnosed with Epithelial Ovarian Cancer:

Study involving serial serum samples from 72 patients (total of 330 pairs of observations).
A positive change in HE4 was defined as an increase in the value that was at least 18% greater than the previous value.
30/61 (49%) of patient samples with a positive change correlated with disease progression.
214/269 (80%) of patient serial samples with no significant change in HE4 value correlated with no progression.
Total concordance: 74% (245/330).

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Monitoring of Disease status in Patients Diagnosed with Epithelial Ovarian Cancer:

Sensitivity: 49.2% (Lower 95% CI: 27.8%, Upper 95% CI: 70.6%)
Specificity: 79.6% (Lower 95% CI: 65.9%, Upper 95% CI: 93.2%)
Total Concordance: 73.9% (Lower 95% CI: 62.6%, Upper 95% CI: 85.3%)
PPV: 35.3% (Lower 95% CI: 19.9%, Upper 95% CI: 50.7%)
NPV: 87.4% (Lower 95% CI: 72.3%, Upper 95% CI: 102.4%)

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

Fujirebio Diagnostics, Inc. HE4 EIA, K072939

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

Regulation Number and Section

§ 866.6010 Tumor-associated antigen immunological test system.

(a)
Identification. A tumor-associated antigen immunological test system is a device that consists of reagents used to qualitatively or quantitatively measure, by immunochemical techniques, tumor-associated antigens in serum, plasma, urine, or other body fluids. This device is intended as an aid in monitoring patients for disease progress or response to therapy or for the detection of recurrent or residual disease.(b)
Classification. Class II (special controls). Tumor markers must comply with the following special controls: (1) A guidance document entitled “Guidance Document for the Submission of Tumor Associated Antigen Premarket Notifications (510(k)s) to FDA,” and (2) voluntary assay performance standards issued by the National Committee on Clinical Laboratory Standards.

Summary

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract symbol that resembles a stylized caduceus or a human figure with outstretched arms.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

January 8, 2016

Fujirebio Diagnostics, Inc. c/o Stacey Dolan, RAC-US Manager, Regulatory Affairs 201 Great Valley Parkway Malvern, PA 19355

Re: K151378

Trade/Device Name: Lumipulse G HE4 Immunoreaction Cartridges, Lumipulse G HE4 Calibrators Regulation Number: 21 CFR 866.6010 Regulation Name: Tumor-associated antigen immunological test system Regulatory Class: II Product Code: OIU, JIT Dated: October 14, 2015 Received: October 15, 2015

Dear Ms. Dolan:

This letter corrects our substantially equivalent letter of November 23, 2015.

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate 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) that do not require approval of a premarket approval application (PMA). 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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the

1

electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Kelly Oliner -S

FOR

Leonthena R. Carrington, MS, MBA, MT(ASCP) Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K151378

Device Name

Lumipulse G HE4 Immunoreaction Cartridges Lumipulse G HE4 Calibrators

Indications for Use (Describe) Lumipulse G HE4 Immunoreaction Cartridges:

For in vitro diagnostic use.

Lumipulse G HE4 is a Chemiluminescent Enzyme Immunoassay (CLEIA) for the quantitative determination of HE4 in human serum and plasma (lithium heparin or dipotassium EDTA) on the Lumipulse G System.

The assay is to be used as an aid in monitoring recurrence or progressive disease in patients with epithelial ovarian cancer. Serial testing for patient HE4 values should be used in conjunction with other clinical methods used for monitoring epithelial ovarian cancer.

Lumipulse G HE4 Calibrators:

Lumipulse G HE4 Calibrators are for use in the calibration of the Lumipulse G System for the quantitative measurement of HE4 in human serum and plasma (lithium heparin or dipotassium EDTA).

Type of Use (Select one or both, as applicable)
Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

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Image /page/3/Picture/1 description: The image shows the logo for Fujirebio Diagnostics, Inc. The logo consists of a blue and white abstract symbol on the left, followed by the company name in blue text. The word "FUJIREBIO" is in a larger, bolder font than "Diagnostics, Inc.", which is placed below it. The overall design is clean and professional.

Section 5 510(k) SUMMARY

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

A. 510(k) Number:

K151378

B. Purpose for Submission:

New device

C. Measurand:

HE4

D. Type of Test:

Quantitative assay, automated chemiluminescent enzyme immunoassay (CLEIA)

E. Applicant:

| Address: | Fujirebio Diagnostics, Inc.
201 Great Valley Parkway
Malvern, PA 19355 |
|-----------------|------------------------------------------------------------------------------|
| Contact person: | Stacey Dolan, RAC (US)
(610) 240-3917
dolans@fdi.com |

Summary preparation date: May 21, 2015

F. Proprietary and Established Names:

Lumipulse G HE4 Immunoreaction Cartridges Lumipulse G HE4 Calibrators

G. Regulatory Information:

1. Regulation section: 21 CFR § 866.6010, Tumor-associated antigen immunological test system 21 CFR § 862.1150, Calibrator
1. Classification: Class II
1. Product code: OIU, Test, Epithelial Ovarian Tumor-Associated Antigen (HE4) JIT, Calibrator, Secondary

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Image /page/4/Picture/1 description: The image shows the logo for Fujirebio Diagnostics, Inc. The logo consists of a stylized blue and green graphic on the left, followed by the company name in blue text. The word "FUJIREBIO" is in a larger, bolder font than "Diagnostics, Inc.", which is placed below it. The overall design is clean and professional.

1. Panel:
  82, Immunology (Assay) 75, Chemistry (Calibrators)

H. Intended Use:

1. Intended use(s): See indications for use below.
1. Indication(s) for use:

Lumipulse G HE4 Immunoreaction Cartridges For in vitro diagnostic use.

The Lumipulse G HE4 is a Chemiluminescent Enzyme Immunoassay (CLEIA) for the quantitative determination of HE4 in human serum and plasma (lithium heparin or dipotassium EDTA) on the LUMIPULSE G System.

The assay is to be used as an aid in monitoring recurrence or progressive disease in patients with epithelial ovarian cancer. Serial testing for patient HE4 assay values should be used in conjunction with other clinical methods used for monitoring epithelial ovarian cancer.

Lumipulse G HE4 Calibrators

The Lumipulse G HE4 Calibrators are for use in the calibration of the LUMIPULSE G System for the quantitative measurement of HE4 in human serum and plasma (lithium heparin or dipotassium EDTA).

1. Special conditions for use statement(s): Prescription use only
1. Special instrument requirements: LUMIPULSE G1200 System

-Device Description:

The Lumipulse G HE4 is an assay system, including a set of immunoassay reagents, for the quantitative measurement of HE4 in specimens based on CLEIA technology by a two-step sandwich immunoassay method on the LUMIPULSE G System.

Lumipulse G HE4 Immunoreaction Cartridges |REF| 234174

The Lumipulse G HE4 Immunoreaction Cartridges consists of 3 × 14 tests. Each kit contains the following:

Antibody-Coated Particle Solution (Liquid when used, 250 µL/Immunoreaction Cartridge)

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Image /page/5/Picture/1 description: The image contains the logo for Fujirebio Diagnostics, Inc. The logo consists of a stylized graphic on the left, featuring blue and white shapes, next to the company name in blue, bold letters. Below the company name, the words "Diagnostics, Inc." are written in a smaller, thinner font.

Contains 150 ug/mL anti-HE4 monoclonal antibody (mouse)-coated particles, protein stabilizers (bovine and mouse) and chemical stabilizers in 0.15 M sodium chloride/Tris buffer. This solution contains gelatin and turns into gel at 15 °C or lower. Preservative: Proclin 300.

Enzyme-Labeled Antibody Solution

(Liquid, 350 µL/Immunoreaction Cartridge)

Contains 0.25 µg/mL alkaline phosphatase (ALP: calf) labeled anti-HE4 monoclonal antibody (mouse), protein stabilizers (bovine, calf and mouse) and chemical stabilizers in 0.1 M sodium chloride/MES buffer. Preservative: Proclin 300.

LUMIPULSE G HE4 Calibrators CAL 234181

Each calibrator kit contains one bottle each of Calibrators 1 and 2. The calibrator kit is packaged separately.

CAL Liquid, 1 × 2 Concentrations

CAL 1 0 pM HE4 calibrator (1 × 1.5 mL)

CAL 2 1500 pM HE4 calibrator (1 × 1.5 mL)

Contains sodium chloride and potassium chloride in phosphate buffer with protein stabilizer (bovine). Preservative: Proclin 950.

J. Substantial Equivalence Information:

1. Predicate device name(s): Fujirebio Diagnostics, Inc. HE4 EIA
1. Predicate 510(k) number(s): K072939

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Image /page/6/Picture/1 description: The image contains the logo for Fujirebio Diagnostics, Inc. The logo features a stylized graphic element on the left, consisting of blue and green shapes that resemble a water droplet or a stylized leaf. To the right of the graphic is the company name, "FUJIREBIO," in a bold, sans-serif blue font. Below the company name is the text "Diagnostics, Inc." in a smaller, sans-serif font, also in blue.

3. Comparison with predicate:

Similarities
	Lumipulse G HE4 Assay
(Proposed Device)	Fujirebio Diagnostics, Inc. HE4
EIA Assay
(Predicate Device) K072939
Device Type	In vitro diagnostic	In vitro diagnostic
Classification	Class II	Class II
CFR section	866.6010	866.6010
Product Code	OIU	OIU
Product Usage	Clinical and Hospital laboratories
For in vitro diagnostic use.	Clinical and Hospital laboratories
For in vitro diagnostic use.
Intended Use	The Lumipulse G HE4 is a
Chemiluminescent Enzyme
Immunoassay (CLEIA) for the
quantitative determination of HE4 in
human serum and plasma (lithium
heparin or dipotassium EDTA) on the
LUMIPULSE G System.

The assay is to be used as an aid in
monitoring recurrence or progressive
disease in patients with epithelial
ovarian cancer. Serial testing for
patient HE4 assay values should be
used in conjunction with other clinical
methods used for monitoring epithelial
ovarian cancer. | The HE4 EIA is an enzyme
immunometric assay for the
quantitative determination of HE4 in
human serum.

The assay is to be used as an aid in
monitoring recurrence or progressive
disease in patients with epithelial
ovarian cancer. Serial testing for
patient HE4 assay values should be
used in conjunction with other clinical
methods used for monitoring epithelial
ovarian cancer |
| Specimen Collection
Method | Routine Phlebotomy Techniques | Routine Phlebotomy Techniques |
| Calibrator Antigen | Ig-HE4 | Ig-HE4 |
| Analyte | Human HE4 | Human HE4 |

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Image /page/7/Picture/1 description: The image shows the logo for Fujirebio Diagnostics, Inc. The logo features a stylized graphic on the left, consisting of blue and green shapes. To the right of the graphic, the word "FUJIREBIO" is written in large, blue, sans-serif font. Below "FUJIREBIO", the words "Diagnostics, Inc." are written in a smaller, black, sans-serif font.

Differences
	Lumipulse G HE4 Assay
(Proposed Device)	Fujirebio Diagnostics, Inc. HE4
EIA Assay
(Predicate Device) K072939
Instrument System	LUMIPULSE G1200 System	None
Principle of Operation	Chemiluminscent Enzyme
Immunoassay (CLEIA)	Enzyme Immunometric Assay
(EIA)
Type of Specimen	Human Serum or Plasma (, lithium
heparin, or dipotassium EDTA)	Human Serum
Assay Range	20 - 1500 pM	15 - 900 pM
Interpretation of Results	Calibrator Curve	Standard Curve
Calibrators	2 Levels (0 and 1500 pM)
Ready to Use
-Liquid
-Supplied as separate kit	5 Levels (Lot specific)
-Lyophilized
Antibodies	12A2 and 3D8 (mouse
monoclonal)	2H5 and 3D8 (mouse monoclonal)

K. Standard/Guidance Document Referenced (if applicable):

. ISO 17511:2003 Measurement of Quantities in Biological Samples - Metrological Traceability of Values Assigned to Calibrator and Control Materials
. CLSI EP5-A3 - Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline - Third Edition
CLSI EP7-A2 Interference Testing in Clinical Chemistry; Approved Guideline-Second . Edition
. CLSI C28-A3c - Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory: Approved Guideline-Third Edition
. CLSI EP17-A2 - Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline Second Edition
. CLSI EP6-A - Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline
. CLSI EP9-A3 - Measurement Procedure Comparison and Bias Estimation Using Patient Samples; approved Guideline - Third Edition
FDA Guidance Document for the Submission of Tumor Associated Antigen Premarket ● Notifications, [510(k)], to FDA
. Guidance on Informed Consent for In Vitro Diagnostic Device Studies Using Leftover Human Specimens that are Not Individually Identifiable - Guidance for Sponsors, Institutional Review Boards, Clinical Investigators and FDA Staff

L. Test Principle:

Lumipulse G HE4 is an assay system, including a set of immunoassay reagents, for the quantitative measurement of HE4 in specimens based on CLEA technology by a two-step sandwich immunoassay method on the LUMIPULSE G1200 System. HE4 in specimens

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Image /page/8/Picture/1 description: The image is a logo for Fujirebio Diagnostics, Inc. The logo features a stylized blue and white graphic on the left, resembling a water droplet or a stylized letter. To the right of the graphic, the word "FUJIREBIO" is written in a bold, blue sans-serif font. Below "FUJIREBIO", the words "Diagnostics, Inc." are written in a smaller, dark gray sans-serif font.

specifically binds to anti-HE4 monoclonal antibody (mouse) on the particles, and antigenantibody immunocomplexes are formed. The particles are washed and rinsed to remove unbound materials. Alkaline phosphatase (ALP: calf)-labeled anti-HE4 monoclonal antibody (mouse) specifically binds to HE4 of the immunocomplexes on the particles, and additional immunocomplexes are formed. The particles are washed and rinsed to remove unbound materials. Substrate Solution is added and mixed with the particles. AMPPD contained in the Substrate Solution is dephosphorylated by the catalysis of ALP indirectly conjugated to particles. Luminescence (at a maximum wavelength of 477 nm) is generated by the cleavage reaction of dephosphorylated AMPPD. The luminescent signal reflects the amount of HE4.

M. Performance Characteristics (if/when applicable):

Data were generated using the LUMIPULSE G1200 System.

1. Analytical performance:
- a. Precision/Reproducibility:

Lumipulse G HE4 is ≤10% total (within-laboratory) CV (coefficient of variation). Lumipulse G HE4 demonstrated precision ≤3.5% (total %CV) in a study run according to the Clinical and Laboratory Standards Institute (CLSI) guideline EP5-A3. Eight human serum-based samples (specimen pools), and two commercially available serum-based controls were assayed in replicates of two at two separate times of the day for 20 for each sample) using one LUMIPULSE G1200 System. Data from this study are presented below.

| Sample | Mean
Conc.
(pM)
n=80 | Within-run
SD
(pM) | Within-
run
%CV | Total
SD
(pM) | Total
%CV |
|-----------|-------------------------------|--------------------------|-----------------------|---------------------|--------------|
| Serum 1 | 58.9 | 0.70 | 1.2 | 1.15 | 1.9 |
| Serum 2 | 77.4 | 0.78 | 1.0 | 1.45 | 1.9 |
| Serum 4 | 119.0 | 1.58 | 1.3 | 1.87 | 1.6 |
| Serum 5 | 310.7 | 3.18 | 1.0 | 7.12 | 2.3 |
| Serum 6 | 1125.1 | 14.03 | 1.2 | 24.19 | 2.1 |
| Serum 7 | 28.6 | 0.35 | 1.2 | 0.99 | 3.5 |
| Serum 8 | 627.8 | 7.06 | 1.1 | 8.49 | 1.4 |
| Serum 9 | 1329.9 | 21.89 | 1.6 | 29.88 | 2.2 |
| Control 1 | 95.1 | 1.99 | 2.1 | 2.27 | 2.4 |
| Control 2 | 751.8 | 10.27 | 1.4 | 13.88 | 1.8 |

Lumipulse G HE4 demonstrated precision ≤6.1% total %CV when tested at 3 external laboratory sites. The human serum-based samples (specimen pools) and two commercially available serum-based controls were assayed in replicates of two separate times of the day at each of the sites for 10 days (n=40 for each sample) using one LUMIPUSE G1200 System. Data from this study are presented below.

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Image /page/9/Picture/1 description: The image shows the logo for Fujirebio Diagnostics, Inc. The logo consists of a stylized graphic on the left and the company name on the right. The graphic is composed of blue and green shapes, with a white teardrop shape in the center. The text "FUJIREBIO" is in blue, and "Diagnostics, Inc." is in black.

| Sample | Mean
Conc.
(pM)
n=120 | Within-
run
SD (pM) | Within-
run
%CV | Total
SD
(pM) | Total
%CV |
|-----------|--------------------------------|---------------------------|-----------------------|---------------------|--------------|
| Serum 1 | 57.1 | 0.70 | 1.2 | 2.14 | 3.8 |
| Serum 2 | 75.4 | 0.89 | 1.2 | 2.78 | 3.7 |
| Serum 4 | 115.9 | 1.47 | 1.3 | 4.85 | 4.2 |
| Serum 5 | 303.0 | 2.85 | 0.9 | 15.44 | 5.1 |
| Serum 6 | 1110.2 | 14.99 | 1.4 | 62.98 | 5.7 |
| Serum 7 | 27.5 | 0.35 | 1.3 | 1.68 | 6.1 |
| Serum 8 | 609.3 | 8.17 | 1.3 | 32.89 | 5.4 |
| Serum 9 | 1323.2 | 25.92 | 2.0 | 68.74 | 5.2 |
| Control 1 | 95.4 | 1.90 | 2.0 | 5.57 | 5.8 |
| Control 2 | 733.4 | 12.27 | 1.7 | 29.16 | 4.0 |

Lumipulse G HE4 demonstrated precision ≤3.2% total %CV when tested using 3 lots of Lumipulse G HE4 immunoreaction cartridges and calibrators. The human serum-based samples (specimen pools) and two commercially available serum-based controls were assayed in replicates of two at two separate times of the day for each of the lots for 10 days (n=40 for each sample) using one LUMIPULSE G1200 System. Data from this study are presented below.

| Sample | Mean
Conc.
(pM)
n=120 | Within-
run
SD (pM) | Within-
run
%CV | Total
SD (pM) | Total
%CV |
|-----------|--------------------------------|---------------------------|-----------------------|------------------|--------------|
| Serum 1 | 57.5 | 0.56 | 1.0 | 1.24 | 2.2 |
| Serum 2 | 75.3 | 0.76 | 1.0 | 1.75 | 2.3 |
| Serum 4 | 116.1 | 1.42 | 1.2 | 2.71 | 2.3 |
| Serum 5 | 302.7 | 3.81 | 1.3 | 9.76 | 3.2 |
| Serum 6 | 1119.3 | 15.36 | 1.4 | 24.91 | 2.2 |
| Serum 7 | 27.6 | 0.35 | 1.2 | 0.75 | 2.7 |
| Serum 8 | 617.5 | 9.66 | 1.6 | 13.83 | 2.2 |
| Serum 9 | 1334.0 | 25.33 | 1.9 | 33.60 | 2.5 |
| Control 1 | 94.9 | 1.66 | 1.8 | 2.20 | 2.3 |
| Control 2 | 746.5 | 8.35 | 1.1 | 11.82 | 1.6 |

b. Linearity/assay reportable range:

Lumipulse G HE4 demonstrated linearity in a study consistent with the quidelines in the CLSI Protocol EP6-A. One human serum specimen pool and one dipotassium EDTA plasma specimen pool with high HE4 levels were diluted with one human serum specimen pool and one dipotassium EDTA plasma specimen pool with low HE4 levels throughout the range of the assay. The linearity was found in the range of 20.0 to 1500.0 pM. Lumipulse G HE4 correlated with expected concentrations according to the linear regression formulas:

Serum:

Plasma: y= 1.0104 (x) + 12.4869; R-squared: 1.0000

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Image /page/10/Picture/1 description: The image is a logo for Fujirebio Diagnostics, Inc. The logo consists of a blue and white abstract shape on the left, followed by the company name in blue, and the words "Diagnostics, Inc." in black below the company name. The abstract shape is made up of a blue curved shape, a white teardrop shape, and green vertical lines.

Lumipulse G HE4 recovery is 100 ± 10%, ranging from 91% to 107%. A study was performed where known concentrations of HE4 antigen were added to human serum and dipotassium EDTA plasma samples with low endogenous HE4 levels. The concentration of HE4 was determined using the Lumipulse G HE4 and the resulting percent recovery was calculated.

| Sample | Target
HE4
Levels
(pM) | Measured
Conc.
(pM)
(n=3) | Expected
Conc.
(pM) | %
Recovery |
|-------------|----- | Serum
1 | 80 | | 150 | | 450 | | 750 | | 1050 | | 1350 | Serum
2 | 80 | | 150 | | 450 | | 750 | | 1050 | | 1350 | Serum
3 | 80 | | 150 | | 450 | | 750 | | 1050 | | 1350 | Plasma
1 | 80 | | 150 | | 450 | | 750 | | 1050 | | 1350 | Plasma
2 | 80 | | 150 | | 450 | | 750 | | 1050 | | 1350 | Plasma
3 | 80 | | 150 | | 450 | | 750 | | 1050 | | 1350 ----------------------------|------------------------------------|---------------------------|---------------|
| 132.8 | 125.2 | 106 |
| 188.2 | 186.7 | 101 |
| 484.4 | 474.2 | 102 |
| 768.0 | 761.0 | 101 |
| 1056.5 | 1064.4 | 99 |
| 1350.4 | 1390.5 | 97 |
| 148.3 | 140.8 | 105 |
| 196.0 | 191.5 | 102 |
| 488.7 | 479.0 | 102 |
| 767.2 | 765.8 | 100 |
| 1046.9 | 1069.2 | 98 |
| 1316.9 | 1395.3 | 94 |
| 116.8 | 109.3 | 107 |
| 202.4 | 197.4 | 103 |
| 491.7 | 484.9 | 101 |
| 763.8 | 771.7 | 99 |
| 1081.1 | 1075.1 | 101 |
| 1324.1 | 1401.2 | 94 |
| 127.5 | 122.2 | 104 |
| 242.4 | 240.4 | 101 |
| 518.1 | 527.9 | 98 |
| 796.3 | 814.7 | 98 |
| 1069.3 | 1118.1 | 96 |
| 1366.4 | 1444.2 | 95 |
| 138.5 | 134.5 | 103 |
| 206.5 | 203.4 | 102 |
| 492.7 | 490.9 | 100 |
| 780.6 | 777.7 | 100 |
| 1035.8 | 1081.1 | 96 |
| 1274.7 | 1407.2 | 91 |
| 137.9 | 130.9 | 105 |
| 204.8 | 201.1 | 102 |
| 494.0 | 488.6 | 101 |
| 784.3 | 775.4 | 101 |
| 1057.1 | 1078.8 | 98 |
| 1344.9 | 1404.9 | 96 |

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Image /page/11/Picture/1 description: The image shows the logo for Fujirebio Diagnostics, Inc. The logo features a stylized blue and white graphic on the left, resembling a water droplet or a stylized letter 'F'. To the right of the graphic is the company name, "FUJIREBIO" in bold blue letters, with "Diagnostics, Inc." written in a smaller, black font below it. The logo is clean and professional, conveying a sense of innovation and reliability.

High dose hook is a phenomenon whereby very high level specimens may read within the dynamic range of the assay. For Lumipulse G HE4, no high dose hook effect was observed for samples containing up to 300,000 pM of HE4 antigen. However values above 30,000 pM should be interpreted with caution because reading may be inaccurate above this concentration.

Traceability, Stability, Expected values (controls, calibrators, or methods): ﯼ

The Lumipulse G HE4 Calibrators are for use in the calibration of the LUMIPULSE G System when used for the quantitative measurement of HE4 in human serum and plasma (lithium heparin or dipotassium EDTA). 2 Bottles (1.5 mL each) are supplied for the LUMIPULSE G HE4 Calibrators. Preservatives: Proclin 950. The calibrators are at the following concentrations:

Calibrator	Concentration (pM)
CAL 1	0
CAL 2	1500

There is currently no known internationally recognized consensus reference method or reference material for standardization. Lumipulse G HE4 values are expressed as pM (or pmol/L). This value is related to a Fujirebio Diaqnostics maintained reference preparation. Calibration of the Lumipulse G HE4 is traceable to in-house reference calibrators, whose values have been assigned to correlate to Fujirebio Diagnostics' HE4 EIA.

The real time stability (including open use) has been demonstrated for 10 months. The instrument cartridge and calibrators were found to be stable under simulated transport. conditions. Shelf life assignment for the Lumipulse G HE4 Immunoreaction Cartridges and Calibrators will be 8 months at 2-10°C. .

d. Detection limit:

The Limit of Blank (LoB), Limit of Detection (LoD) and Limit of Quantitation (LoQ) of Lumipulse G HE4 is ≤ 20.0 pM.

1) LoB and LoD

The LoB for Lumipulse G HE4 was 0.1 pM.

The LoD for Lumipulse G HE4 was 3.5 pM, determined consistent with the quidelines in the CLSI Protocol EP17-A2. Seven low-level specimens were tested over 3 days using three LUMIPULSE G Systems and three Lumipulse G HE4 lots giving 180 determinations for each panel.

2) LoQ

The LOQ for Lumipulse G HE4 was 3.5 pM, determined consistent with the guidelines in the CLSI quideline EP17-A2. Since the percent total error estimate is ≤ 30%, LoQ equals LoD.

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Image /page/12/Picture/1 description: The image shows the logo for Fujirebio Diagnostics, Inc. The logo consists of a blue and white abstract graphic on the left, followed by the company name in blue and black text on the right. The word "FUJIREBIO" is in a bold, sans-serif font, and the words "Diagnostics, Inc." are in a smaller, lighter font below.

e. Analytical specificity:

The Lumipulse G HE4 demonstrated an average interference of ≤ 10% (for each compound) in a study consistent with the guidelines in the CLSI guideline EP7-A2. Human serum specimens were supplemented with potentially interfering compounds. TThe following compounds were tested and found not to interfere with the test.

| Endogenous
Interferences | Test
Concentration |
|------------------------------------------|-----------------------|
| Free Bilirubin (unconjugated) | 60 mg/dL |
| Conjugated Bilirubin | 60 mg/dL |
| Triglycerides (Intralipid 20% Emulsion) | 3000 mg/dL |
| Hemoglobin | 500 mg/dL |
| Human Serum Albumin | 12 g/dL |
| Immunoglobulin G (IgG) | 5 g/dL |
| Biotin | 19.8 mg/dL |
| Human Anti-Mouse Antibodies (HAMA) | 1,000 ng/mL |
| Rheumatoid Factor (RF) | 1,000 IU/mL |
| Therapeutic drug
Interferences | Test
Concentration |
| Bevacizumab | 750 µg/mL |
| Carboplatin | 600 µg/mL |
| Cisplatin | 180 µg/mL |
| Clotrimazole | 0.3 µg/mL |
| Cyclophosphamide | 800 µg/mL |
| Dexamethansone | 20 µg/mL |
| Docetaxel | 10.5 mg/dL |
| Doxil® (Pegylated Liposomal Doxorubicin) | 5.2 mg/dL |
| Doxorubicin | 120 µg/mL |
| Erlotinib | 150 ug/mL |
| Etoposide | 10 ug/mL |
| 5-Fluorouracil | 900 ug/mL |
| Gemcitabine | 100 ug/mL |
| Leucovorin | 750 µg/mL |
| Magestrol Acetate | 10 ug/mL |
| Melphalan | 15 µg/mL |
| Methotrexate | 450 µg/mL |
| Olaparib | 28.1 mg/dL |
| Paclitaxel | 3.5 ng/mL |
| Rituximab | 750 µg/mL |
| Tamoxifen | 60 ug/mL |
| Topotecan | 0.155 mg/dL |
| Trastuzumab | 600 ug/ml |

f. Assay cut-off:

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Image /page/13/Picture/1 description: The image contains the logo for Fujirebio Diagnostics, Inc. The logo consists of a blue and white abstract symbol on the left, followed by the company name in blue, with "Diagnostics, Inc." in a smaller font size below "FUJIREBIO". The overall design is clean and corporate.

See Clinical Cutoff in M (4) below

2. Comparison studies:

a. Method Comparison

Lumipulse G HE4 method comparison study was performed using specimens consistent with CLSI guideline EP9-A3. The weighted Deming regression method was used to compare Lumipulse G HE4 to HE4 EIA. The HE4 EIA has a different measuring range than Lumipulse G HE4, therefore the results were limited to the measuring range of both devices (143 samples). The data are summarized in the following table.

Range of samples: 35.2 - 969.5 pM (Lumipulse G HE4) 33.4 - 899.0 pM (HE4 EIA)

Lumipulse G HE4 vs. HE4 EIA
n	Correlation
Coefficient
(r)	Intercept
(95% CI)	Slope
(95% CI)	Average Bias
(pM)
143	0.9891	-3.2350	1.0349	8.8
		-5.3628 –	1.0074 –
		–1.1071	1.0624

The date summarized in the following table include results from a study with specimens above the measurement range of both devices requiring dilution (168 samples).

Range of samples:

35.2 - 4602.0 pM (Lumipulse G HE4) 33.4 - 4067.0 pM (HE4 EIA)

Lumipulse G HE4 vs. HE4 EIA

| n | Correlation
Coefficient
(r) | Intercept
(95% CI) | Slope
(95% CI) | Average Bias
(pM) |
|-----|-----------------------------------|-----------------------|--------------------|----------------------|
| 168 | 0.9917 | -2.8518 | 1.0289 | 13.8 |
| | | -4.8632 -
-0.8404 | 1.0045 -
1.0533 | |

b. Matrix Comparison

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Image /page/14/Picture/1 description: The image is a logo for Fujirebio Diagnostics, Inc. The logo consists of a stylized graphic on the left and the company name on the right. The graphic is a combination of blue and green shapes, with a white teardrop shape in the center. The company name is written in blue, with "Diagnostics, Inc." written in a smaller, black font below the main name.

The Lumipulse G HE4 matrix comparison study was performed to evaluate the difference across tube types (SST, K2EDTA, and Lithium Heparin,) versus the means of the control samples (Red top serum) analyzed per CLSI guideline EP9-A3. The slope for each tube type when compared to the control had 95% confidence intervals that lay entirely within the range 0.9 to 1.1 and the correlation coefficients were ≥ 0.9.

1. Clinical studies:
- a. Clinical sensitivity:

See 3(c) below

b. Clinical specificity:

See 3(c) below

c. Other clinical supportive data (when a. and b. are not applicable):

Monitoring of Disease status in Patients Diagnosed with Epithelial Ovarian Cancer

The effectiveness of Lumipulse G HE4 as an aid in monitoring of disease status in EOC patients was determined by assessing changes in HE4 levels in serial serum samples from 72 patients compared to changes in disease status. A study involving a total of 330 pairs of observations was undertaken with an average number of 5.6 observations per patient. A positive change in HE4 was defined as an increase in the value that was at least 18% greater than the previous value of the test. This level of change takes into account the variability of the assay. Forty-nine percent (49%) or 30/61 of the patient samples with a positive change correlated with the disease progression while eighty percent (80%) or 214/269 of the patient serial samples with no significant change in HE4 value correlated with no progression. The total concordance was seventy-four percent (74% or 245/330).

| Performance Measurement | SE | Lower 95%
CI | Upper 95%
CI | |
|-------------------------|------|-----------------|-----------------|-------|
| Sensitivity | 49.2 | 10.74 | 27.8 | 70.6 |
| Specificity | 79.6 | 6.87 | 65.9 | 93.2 |
| Total Concordance | 73.9 | 5.69 | 62.6 | 85.3 |
| PPV | 35.3 | 7.71 | 19.9 | 50.7 |
| NPV | 87.4 | 7.54 | 72.3 | 102.4 |

Lumipulse G HE4 Performance Measurements (%):

SE = Standard Error

Cl = Confidence Interval

The following table presents the data in a 2 x 2 format.

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Image /page/15/Picture/1 description: The image shows the logo for Fujirebio Diagnostics, Inc. The logo consists of a stylized graphic on the left, featuring blue and green elements that resemble a water droplet and vertical lines. To the right of the graphic, the company name "FUJIREBIO" is displayed in bold, blue letters, with "Diagnostics, Inc." appearing in a smaller font size below it.

Change in Disease State per Sequential Pair
Change in
HE4
Concentration	No
progression	Progression	Total
≤ 18%	214	31	245
> 18%	55	30	85
Total	269	61	330

4. Clinical cut-off:

HE4 is significantly elevated if it is at least 18% higher than the previous assay.

5. Expected values/Reference range:

The distribution of HE4 levels determined in healthy premenopausal and postmenopausal women is shown in the table below:

| | All
Healthy Subjects | Premenopausal
Healthy Subjects | Postmenopausal
Healthy Subjects |
|----------------------------------------------------------------|-------------------------|-----------------------------------|------------------------------------|
| N | 240 | 120 | 120 |
| Mean | 59.5 | 58.0 | 61.0 |
| (SD) | (68.8) | (94.8) | (22.5) |
| Median | 51.3 | 47.0 | 55.9 |
| Range (min, max) | 29.4, 1076.6 | 29.9, 1076.6 | 29.4, 176.7 |
| Reference Interval
(2.5th percentile,
97.5th percentile) | 32.5, 108.4 | 31.9, 87.1 | 32.5, 112.2 |
| Proportion below
90.0 pM | 96% | 98% | 93% |
| Proportion below
135.0 pM | 99% | 99% | 98% |

All Lumipulse G HE4 concentrations are presented as pM

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Image /page/16/Picture/1 description: The image shows the logo for Fujirebio Diagnostics, Inc. The logo consists of a stylized graphic on the left, featuring blue and green elements, possibly representing water droplets or abstract shapes. To the right of the graphic, the company name "FUJIREBIO" is displayed in bold, blue letters, with "Diagnostics, Inc." written in a smaller, black font underneath.

The distribution of HE4 levels determined in benign conditions is shown in the table below:

| | Benign
Gynecological
Disease | Other
Benign
Disease | Congestive
Heart
Failure | Hypertension | Pregnant |
|-----------------------------------------------------------------|------------------------------------|----------------------------|--------------------------------|-----------------|---------------|
| N | 366 | 40 | 40 | 40 | 40 |
| Mean | 70.7 | 232.6 | 185.3 | 397.0 | 45.4 |
| (SD) | (44.0) | (548.8) | (99.7) | (999.9) | (8.3) |
| Median | 57.8 | 81.4 | 173.9 | 108.8 | 44.5 |
| Range (min, max) | 26.3,
386.8 | 42.7,
2762.0 | 30.6,
417.7 | 39.0,
5773.0 | 31.9,
71.0 |
| Reference Interval
(2.5th percentile ,
97.5th percentile) | 35.0,
202.0 | 44.2,
2374.9 | 43.5,
407.5 | 40.9,
2874.3 | 32.2,
58.3 |
| Proportion below
90.0 pmol/L | 82% | 55% | 18% | 43% | 100% |
| Proportion below
135.0 pmol/L | 94% | 70% | 38% | 65% | 100% |

*All Lumipulse G HE4 concentrations are presented as pM

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Image /page/17/Picture/1 description: The image is a logo for Fujirebio Diagnostics, Inc. The logo features a stylized blue and white graphic on the left, resembling a water droplet or a stylized leaf. To the right of the graphic is the company name, "FUJIREBIO" in bold, blue letters, with "Diagnostics, Inc." underneath in a smaller font. The logo is clean and professional, conveying a sense of innovation and reliability.

The distribution of HE4 levels determined in malignancy conditions is shown in the table below:

| | Epithelial
Ovarian
Cancer | Bladder
Cancer | Breast
Cancer | Endometrial
Cancer | GI
Cancer | Lung
Cancer |
|-----------------------------------------------------------------|---------------------------------|-------------------|------------------|-----------------------|-----------------|------------------|
| N | 155 | 40 | 40 | 40 | 40 | 40 |
| Mean
(SD) | 1188.7
(2447.4) | 188.3
(230.0) | 173.0
(398.0) | 145.9
(119.5) | 103.4
(51.8) | 157.6
(127.5) |
| Median | 341.0 | 127.5 | 95.3 | 99.5 | 91.2 | 115.8 |
| Range (min, max) | 24.6,

15000.0 | 44.9,
1381.9 | 32.4,
2591.0 | 50.0,
646.9 | 42.2,
258.9 | 41.1,
658.5 |
| Reference Interval
(2.5th percentile ,
97.5th percentile) | 48.6,
7602.3 | 50.3,
569.9 | 38.3,
419.8 | 60.5,
500.0 | 44.3,
247.7 | 42.3,
562.9 |
| Proportion below
90.0 pmol/L | 14% | 30% | 48% | 35% | 48% | 33% |
| Proportion below
135.0 pmol/L | 25% | 58% | 78% | 68% | 83% | 63% |

All Lumipulse G HE4 concentrations are presented as pM

It is recommended that each laboratory establish its own reference value for the population of interest for Lumipulse G HE4 on the LUMIPULSE G1200.

6. Conclusion

The results of these analytical (nonclinical) and clinical studies demonstrate that the Lumipulse G HE4 assay is substantially equivalent to the performance of the Fujirebio Diagnostics, Inc. HE4 EIA assay.

N. Proposed Labeling:

The labeling satisfies the requirements of 21 CFR Part 809.10.