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 ovarian cancer.

Device Description

The HE4 EIA is a solid-phase, non-competitive immunoassay based upon the direct sandwich technique based on two mouse monoclonal antibodies, 2H5 and 3D8, directed against two epitopes in the C-WFDC domain of HE4. Calibrators, controls and patient samples are incubated together with biotinylated Anti-HE4 monoclonal antibody (MAb) 2H5 in streptavidin coated microstrips. HE4 present in calibrators or samples is adsorbed to the streptavidin coated microstrips by the biotinylated Anti-HE4 MAb during the incubation. The strips are then washed and incubated with HRP labeled Anti-HE4 MAb 3D8. After washing, buffered Substrate/Chromogen reagent (hydrogen peroxide and 3, 3', 5' tetra-methyl-benzidine) is added to each well and the enzyme reaction is allowed to proceed. During the enzyme reaction a blue color will develop if antigen is present. The intensity of the color is proportionate to the amount of HE4 present in the samples. The color intensity is determined in a microplate spectrophotometer at 620 nm (or optionally at 405 nm after addition of Stop Solution). Calibration curves are constructed for each assay by plotting absorbance value versus the concentration for each calibrator. The HE4 concentrations of patient samples are then read from the calibration curve.

AI/ML Overview

The provided text describes the HE4 EIA Kit, its intended use, and performance characteristics for monitoring epithelial ovarian cancer. Here's a breakdown of the information requested:

1. Table of Acceptance Criteria and Reported Device Performance

The document doesn't explicitly state "acceptance criteria" for clinical performance. Instead, it presents the results of a study and lists sensitivity and specificity at various percent changes in HE4 concentration. The study aimed to demonstrate the device's effectiveness as an aid in monitoring recurrence or progressive disease. For the purpose of this table, I will present the reported performance at a 25% change in HE4 concentration, as this was the change used in the main 2x2 table and discussed in the text. Other performance characteristics are listed as "The HE4 EIA Kit precision is ≤ 15% total CV," etc. which can be considered performance goals or criteria.

Performance Characteristic/Acceptance Criteria	Reported Device Performance
Precision (Total CV)	≤ 15%
Linearity (Mean Recovery)	Within 15% of expected
Functional Sensitivity (CV 20%)	< 5 pM
Detection Limit (LoD)	< 2.5 pM
Interference (Assay Specificity)	100% ± 15% recovery
Monitoring Effectiveness (at >25% change)
Sensitivity (Concordance to Progression)	60% (76/126)
Specificity (Concordance to No Progression)	75% (171/228)
Total Concordance	70% (247/354)

2. Sample Size Used for the Test Set and the Data Provenance

Sample Size for Test Set: 80 patients, contributing to a total of 354 pairs of observations (mean of 4.4 observations per patient).
Data Provenance: Not explicitly stated (e.g., country of origin). The study appears to be retrospective based on "serial serum samples" from patients, but this is not explicitly confirmed as prospective or retrospective.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and the Qualifications of Those Experts

The document does not specify the number of experts used or their qualifications for establishing the ground truth ("change in disease status"). It simply refers to "disease status."

4. Adjudication Method for the Test Set

The document does not specify an adjudication method (e.g. 2+1, 3+1, none) for determining the ground truth for the test set.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

This is an immunoassay device, not an imaging or diagnostic device involving human "readers" or AI assistance in the way an MRMC study would typically evaluate. Therefore, no MRMC comparative effectiveness study involving human readers with or without AI assistance was performed or is applicable here. The study compares the device's output to "disease status."

6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was done

Yes, the performance characteristics and monitoring study described are for the device (HE4 EIA Kit) in a standalone capacity, i.e., without a human-in-the-loop performance evaluation in the context of its primary monitoring function. The "Intended Use" states it should be used "in conjunction with other clinical methods," implying it is an aid to a physician's overall assessment, but the performance data presented is for the assay itself.

7. The Type of Ground Truth Used

The ground truth used was "changes in disease status" for ovarian cancer patients. The method for determining "disease status" is not explicitly defined (e.g., pathology, clinical follow-up, imaging results, outcomes data), but it is implied to be a clinical assessment.

8. The Sample Size for the Training Set

The document does not specify a separate "training set" or sample size for training. This is an immunoassay kit, where "training" typically refers to the development and optimization of the assay itself (e.g., antibody selection, reagent concentrations), which is not detailed in terms of patient sample sizes. The performance studies described appear to be validation or verification studies.

9. How the Ground Truth for the Training Set Was Established

As no explicit "training set" with associated ground truth is mentioned in the context of machine learning or algorithm development, this information is not applicable or provided. The ground truth for the performance studies (test set) was based on "changes in disease status," as described in point 7.

Summary

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510(k) SUMMARY

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is: K072939

Submitter Information

JUN - 9 2008

Address:

Fujirebio Diagnostics, Inc. 201 Great Valley Parkway Malvern, PA 19355

Contact person:

Diana L. Dickson (610) 240-3917 FAX (610) 240-3803 dicksond@fdi.com

Summary preparation date: May 13, 2008

Name of Device

Trade/Proprietary Name:	HE4 EIA Kit
-------------------------	-------------

Common/Usual Name: · HE4 EIA Test Kit

21 CFR 866.6010 Requiation Number:

Regulatory Class: Class II

To be determined Product Code:

Predicate Device

ABBOTT ARCHITECT CA 125 II Assay (K042731)

Device Description

The HE4 EIA is a solid-phase, non-competitive immunoassay based upon the direct sandwich technique based on two mouse monoclonal antibodies, 2H5 and 3D8, directed against two epitopes in the C-WFDC domain of HE4. Calibrators, controls and patient samples are incubated together with biotinylated Anti-HE4 monoclonal antibody (MAb) 2H5 in streptavidin coated microstrips. HE4 present in calibrators or samples is adsorbed to the streptavidin coated microstrips by the biotinylated Anti-HE4 MAb during the incubation. The strips are then washed labeled Anti-HE4 MAb 3D8. After washing, buffered and incubated with HRP Substrate/Chromogen reagent (hydrogen peroxide and 3, 3', 5' tetra-methyl-benzidine) is added to each well and the enzyme reaction is allowed to proceed. During the enzyme reaction a blue color will develop if antigen is present. The intensity of the color is proportionate to the

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Image /page/1/Picture/1 description: The image shows the logo for Fujirebio Diagnostics, Inc. The logo consists of a stylized graphic on the left, followed by the company name in bold, black letters. Below the company name, the words "Diagnostics, Inc" are printed in a smaller, non-bold font.

amount of HE4 present in the samples. The color intensity is determined in a microplate spectrophotometer at 620 nm (or optionally at 405 nm after addition of Stop Solution),

Calibration curves are constructed for each assay by plotting absorbance value versus the concentration for each calibrator. The HE4 concentrations of patient samples are then read from the calibration curve.

Reportable Range

The reportable range for the HE4 EIA Kit is 15 - 900 pM

Intended Use

The HE4 ElA 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 ovarian cancer.

Statement of Substantial Equivalence

As there is no FDA cleared or approved device for the HE4 EIA Kit, substantial equivalence for the HE4 EIA test kit was determined by comparing the clinical performance to the ARCHITECT CA 125 II assay as an aid in monitoring patients with epithelial ovarian cancer.

The requlatory submission will be prepared pursuant to Title 21CFR § 866.6010(b) which states Tumor Markers must comply with the following special controls;

1. Guidance document entitled "Guidance Document for the Submission of Tumor Associated Antigen Premarket Notifications (510(k)s) to FDA".
1. Voluntary assay performance standards issued by the National Committee on Clinical Laboratory Standards (NCCLS).

A comparison of the features of the HE4 EIA Kit and the ARCHITECT CA 125 II assay are as follows:

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	Similarities
	HE4 EIA Kit(Proposed Device)	ARCHITECT CA 125 II(Predicate Device)K042731
Device Type	In vitro diagnostic	In vitro diagnostic
Classification	Class II	Class II
Regulation Number	21 CFR 866.6010	21 CFR 866.6010
Product Usage	Clinical and Hospital laboratories	Clinical and Hospital laboratories
Intended Use	Aid in monitoring patients withepithelial ovarian cancer	Aid in monitoring patients withepithelial ovarian cancer
Specimen CollectionMethod	Routine Phlebotomy Techniques	Routine Phlebotomy Techniques
Interpretation of Results	Standard Curve	Standard Curve

	Differences
	HE4 EIA Kit(Proposed Device)	ARCHITECT CA 125 II(Predicate Device)K042731
Product Code	To be determined	LTK
Type of Specimen	Human Serum Only	Human serum or plasma (EDTA,Lithium Heparin, Sodium Heparin)
Antigen Detected	HE4	CA 125
Capture Antibody	2H5 mouse monoclonal	OC 125 mouse monoclonal
Detection Antibody	3D8 mouse monoclonal	M11 mouse monoclonal
Calibrators	6 Levels (0 - 900 pM)Supplied with Kit	6 Levels (0 - 1000 U/mL)Supplied as separate Kit
Controls	2 Levels (50 and 400 pM)Supplied with Kit	3 Levels (40, 300 and 650 U/mL)Supplied as separate Kit
Principle of Operation	ManualEnzymatic Immunoassay (EIA)	AutomatedChemiluminscent MicroparticleImmunoassay (CMIA)

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Performance Characteristics

Precision:

A study was performed with the HE4 EIA Kit modeled after the NCCLS (CLSI) guideline EP5-A2. A panel of four serum samples was assayed, using two lots of reagents, in replicates of two, at two separate times per day for 20 days. The HE4 EIA Kit precision is ≤ 15% total CV.

Linearity:

A dilution linearity study was conducted for the HE4 EIA Kit modeled affer the NCCLS (CLSI) guideline EP6-A. Serum samples with elevated HE4 values were diluted with HE4 Calibrator A (zero). The HE4 concentration was determined for each dilution and the percent (%) recovery was calculated.

The HE4 EIA Kit was determined to have a mean recovery within 15% of the expected result for diluted samples.

Functional Sensitivity:

The functional sensitivity of the HE4 EIA Kit is ≤ 25 pM. The functional sensitivity is expressed as the concentration of an analyte at which the CV is 20%. The NCCLS guideline EP5-A2 was used to design the experiments for determination of functional sensitivity. A study was conducted where a five member sensitivity panel was tested in replicates of 4 in 2 runs on twenty separate days with two lots of reagents. The functional sensitivity determined for the HE4 EIA was found to be < 5 pM.

Detection Limit:

The limit of detection of the HE4 EIA Kit is ≤ 15 pM. The limit of detection (LoD) corresponds to the upper limit of the 95% confidence interval and represents the lowest concentration of HE4 antigen that can be distinguished from zero. The NCCLS guideline EP17-A was used to design the LoD experiments. A study was conducted where HE4 Calibrator A (zero) and 4 samples from healthy subjects diluted to 5 pM with Sample Diluent was tested in replicates of 24 per run in 4 runs on two separate days. The LoD was calculated as follows:

LoD (pM) = 5.0 pM x (1.65 x SDg + 1.65 x SDg) / (ODs - ODg)

The Limit of Detection of the HE4 EIA Kit was calculated to be < 2.5 pM.

Interference:

The HE4 EIA Kit mean assay specificity is 100 ± 15%. Recovery studies were performed to compare sera containing the following compounds at the indicated concentrations with control sera. The NCCLS guideline EP7-A was used to design the interference experiments. The following substances and concentrations were tested and found not to interfere with the test.

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Endogenous serum interferences	Test Concentration
Triglycerides	30 mg/mL
Billirubin	0.2 mg/mL
Hemoglobin	10 mg/mL
Total Protein	120 mg/mL
Chemotherapeutic drug interferences	Test Concentration
Carboplatin	500 µg/mL
Cisplatin	165 µg/mL
Clotrimazole	0.3 µg/mL
Cyclophosphamide	500 µg/mL
Dexamethasone	10 µg/mL
Doxorubicin	1.16 µg/mL
Leucovorin	2.68 µg/mL
Melphalan	2.8 µg/mL
Methotrexate	45 µg/mL
Paclitaxel	3.5 ng/mL

Potentially interfering clinical conditions

The HE4 EIA Kit mean assay specificity is 100 ± 15%. The HE4 EIA Kit was evaluated using specimens with HAMA and Rheumatoid Factor (RF) to further assess the assay specificity. Five specimens positive for HAMA and five specimens positive for RF were evaluated for % recovery with HE4 EIA antigen spiked into each specimen at approximately 50 and 450 pM.

Monitoring of Disease status in Patients Diagnosed with Ovarian Cancer

The effectiveness of the HE4 EIA as an aid in monitoring of disease status in ovarian cancer patients was determined by assessing changes in HE4 levels in serial serum samples from 80 patients compared to changes in disease status. A study involving a total of 354 pairs of observations was undertaken with an average number of 4.4 observations per patient. A positive change in HE4 was defined as an increase in the value that was at least 25% greater than the previous value of the test. This level of change takes into account the variability of the assay and the biological variability. Sixty percent (60%) or 76/126 of the patient samples with a positive change correlated with the disease progression while seventy-five percent (75%) or 171/228 of the patient serial samples with no significant change in HE4 value correlated with no progression. The total concordance was seventy percent (70% or 247/354). The following table presents the data in a 2 x 2 format.

Change in Disease State per Sequential Pair
Increase in HE-4concentration	Progression	No Progression	Total
>25%	76	57	133
<25%	50	171	221
Total	126	228	354

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The following table shows the resulting sensitivities and specificities of the HE4 EIA compared to the disease status at various changes in HE4 EIA concentrations. Sensitivity is represented as a concordance of the HE4 EIA to progression of disease. Specificity is represented as a concordance of the HE4 EIA to no progression of disease.

Percent Change in HE4 Concentration	Sensitivity	Specificity
(%)	(%)	(%)
10	71	62
25	60	75
50	43	88
75	38	92
100	31	95

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Fujirebio Diagnostics, Inc. c/o Ms. Diana L. Dickson Manager Regulatory Affairs 201 Great Valley Parkway Malvern, PA 19355-1308

JUN - 9 2008

Re: K072939

Trade/Device Name: HE4 EIA Regulation Number: 21 CFR 866.6010 Regulation Name: Tumor-associated antigen immunological test system Regulatory Class: Class II Product Code: OIU Dated: May 29, 2008 Received: May 30, 2008

Dear Ms Dickson:

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.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), 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 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 Part 801); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820). This letter will allow you to begin marketing your device as described in your Section 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.

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Page 2 -

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (240) 276-0450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding postmarket surveillance, please contact CDRH's Office of Surveillance and Biometric's (OSB's) Division of Postmarket Surveillance at 240-276-3474. For questions regarding the reporting of device adverse events (Medical Device Reporting (MDR)), please contact the Division of Surveillance Systems at 240-276-3464. You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (240) 276-3150 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Robert H. Becker

Robert L. Becker, Jr., M.D., Ph.D. Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known): K072939

Device Name: HE4 EIA

Indications For Use:

Prescription Use V (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 801 Subpart C)

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

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

Mona M Chan

Division Sign-Off

Office of in Viro Diagnostic Device Evaiuation and Safety Page 1 of 1

510(k): K072939

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.