(207 days)
Not Found
No
The device is a physical control slide for immunohistochemistry staining and does not involve any software or analytical components that would utilize AI/ML.
No.
The device is described as qualitative on-slide controls to monitor the performance of the analytic components of the immunohistochemical staining process for certain HER2, ER, and PR IHC stains. It is not intended for treatment or therapy.
No
The device is described as controls for monitoring the performance of IHC staining, specifically stating "HER2/ER/PR IHControls® are not intended to be used for scoring HER2, ER, and PR IHC stained slides" and "are not intended to replace the controls approved or cleared as part of an IHC device." This indicates it's an assay control, not a diagnostic device for patient samples.
No
The device is a physical product consisting of microbeads in a liquid suspension, intended for use as on-slide controls in immunohistochemistry. It is not software.
Based on the provided information, this device is an In Vitro Diagnostic (IVD).
Here's why:
- Intended Use: The device is intended to "monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process" for specific markers (HER2, ER, PR) in "formalin-fixed paraffin-embedded (FFPE) breast tumor samples." This directly relates to assessing the quality and reliability of a diagnostic test (IHC staining) performed on human specimens.
- Device Description: It is described as "immunostaining positive controls for diagnostic immunohistochemistry laboratories." Controls used in diagnostic tests to ensure the test is working correctly are a key component of IVDs.
- Performance Studies: The submission includes analytical and clinical performance studies demonstrating the device's specificity, reproducibility, and concordance with conventional controls. These types of studies are required for IVD devices to demonstrate their performance characteristics.
- Predicate Device: A predicate device (K023335; QCS HER2 Immunocontrols) is listed, which is also an immunocontrol, further indicating that this type of device is regulated as an IVD.
While the device itself is not used for scoring patient samples, its function is to ensure the accuracy and reliability of the IHC staining process, which is a diagnostic procedure. Therefore, it falls under the definition of an IVD.
N/A
Intended Use / Indications for Use
HER2/ER/PR IHControls® -Level H are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® -Level H are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level H are an additional controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® -Level M are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® -Level M are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level M are an additional controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® -Level L are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® -Level L are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level L are an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
Product codes (comma separated list FDA assigned to the subject device)
NJW
Device Description
HER2/ER/PR IHControls® - Level H, HER2/ER/PR IHControls® - Level M and HER2/ER/PR IHControls® - Level L (HER2/ER/PR IHControls®) are immunostaining positive controls for diagnostic immunohistochemistry laboratories. The same immunohistochemical reaction that occurs on a tissue or cellular sample also occurs on the HER2/ER/PR IHControls® microbead. As a result of immunostaining, the microbead bearing the analyte turns the same color as cells expressing the analyte.
HER2/ER/PR IHControls® incorporate assay controls for human epidermal growth factor receptor type II (HER-2), estrogen receptor (ER), and progesterone receptor (PR). The HER2/ER/PR panel includes controls at 3 different analyte concentrations, providing users with an opportunity to select the control with an analyte concentration within the dynamic range of their assay.
HER2/ER/PR IHControls® are provided in a small vial containing enough liquid suspension for at least 100 tests. The user is instructed to pipet a one microliter droplet onto the slide that also bears the patient sample. The droplet is a suspension containing microscopic glass microbeads that are approximately the same size as cells. The microbead surface bears the molecule(s) being measured. Peptides representing the epitopes of all of the major commercial HER2. ER. and PR antibodies used by clinical immunohistochemistry laboratories are incorporated into the products.
HER2/ER/PR IHControls® are comprised of two different microbeads: analyte-coated glass test microbeads (7-8 um in diameter) and color standard microbeads (4.5 um in diameter). The latter provide a fixed brown color intensity for use in standardizing stain intensity measurement.
The HER2/ER/PR IHControls® panel is provided at 3 different analyte concentrations. In descending order of concentration, these levels are denoted "H", "M", and "L". These different products allow matching the analyte concentration to the dynamic range of the laboratory's stain.
- Level H (high): This product includes analytes for HER2, ER, and PR, at approximately 10^6 molecules per microbead.
- Level M (medium): This product includes analytes for HER2, ER, and PR, at approximately 10^5 molecules per microbead.
- Level L (low): This product includes analytes for HER2, ER, and PR, at approximately 10^4 molecules per microbead.
These concentrations are approximate; they are not intended as assayed controls.
Three product levels are provided to allow the user to select a control with the lowest analyte concentration that still produces an easily visible immunostain.
The immunoreactivity pattern for product levels H, M, and L is described in Table 1. The table indicates the primary antibodies corresponding with up to 9 separate HER2/ER/PR peptides.
Users pipette a droplet onto a microscope slide that also bears a patient's tissue sample. The droplet hardens upon drying, adheres the analyte-coated microbeads to the microscope slide, and is then processed with the patient sample. The hardened droplet is able to pass through dry heat associated with "baking" of slides, organic solvents associated with deparaffinization, boiling associated with antigen retrieval, and repeated rinses associated with immunostaining.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
breast tumor samples
Indicated Patient Age Range
Not Found
Intended User / Care Setting
diagnostic immunohistochemistry 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)
Analytical Performance Studies
Specificity
The HER2/ER/PR IHControls® were tested against twenty-six (26) antigenically irrelevant, commonly used primary antibodies, in the context of clinical IHC testing. Positive tissue controls demonstrated that the antigenically irrelevant tests were operating correctly. No cross-reactivity was detected with the HER2/ER/PR IHControls®. When an appropriate ER, HER2 and PR primary antibody was used for staining, HER2/ER/PR IHControls® demonstrated immunoreactivity.
Shelf Life
The shelf life for each HER2/ER/PR IHControl® level, when stored at 2 - 8 ° C, was tested in real time over approximately 2 years. Shelf life for Level H (BRLS11), Level M (BRL2U04), and Level L (BRL2W08) is 392 days.
Reproducibility of Pathologist IHControl® Readouts
Three pathologists evaluated a blinded slide set of 30 stained HER2/ER/PR IHControl® slides. Ten (10) slides each were stained for HER2, ER, and PR. In each set of ten, 4 were stained with a diluted primary antibody, or no primary antibody, to create weak or absent staining. Overall agreement among pathologists for the interpretation of HER2/ER/PR IHControls® was 100%. Pathologists are able to visually interpret HER2/ER/PR IHControls® and reproducibly render Pass or Fail scores.
Clinical Performance Studies
The HER2/ER/PR IHControls® were incorporated as on-slide controls for HER2, ER, and/or PR immunostaining at three clinical immunohistochemical laboratories. The laboratories incorporated the HER2/ER/PR IHControls® but otherwise followed their typical method for patient testing. After immunostaining was complete, the laboratories performed an assessment. For each stained slide, the pathologist interpreted both the conventional (tissue) control and the HER2/ER/PR IHControls®, for each of the HER2, ER, and PR stains. The controls (tissue controls or HER2/ER/PR IHControls®) were scored as either "Pass" or "Fail", depending on whether the control shows that the test passed or failed the quality control check. Percent concordance is reported.
The overall concordance rate is 440 out of 442 tests, or 99.5%, from all 3 clinical trial sites, for ER, PR, and HER2 tests combined. Individual site concordance was 100%, 99.5%, and 99.1%.
Two discrepancies between tissue controls and HER2/ER/PR IHControls® were due to operator error: (1) use of an incorrect HER2/ER/PR IHControl®, and (2) placement of the HER2/ER/PR IHControls® at the slide edge because the patient sample and tissue (comparator) control left no other place. There was also one site where a histotechnologist did not pipette the HER2/ER/PR IHControls® onto the slide.
The data demonstrate that the HER2/ER/PR IHControls® provide an equally accurate assessment of IHC test quality.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Specificity was tested against 26 antigenically irrelevant primary antibodies, yielding no cross-reactivity.
Reproducibility of Pathologist IHControl® Readouts: Overall agreement among pathologists was 100%.
Clinical Performance Studies: Overall concordance rate between tissue controls and HER2/ER/PR IHControls® was 99.5% (440/442) across 3 clinical sites. Individual site concordance rates were 100%, 99.5%, and 99.1%.
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.
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
§ 864.1860 Immunohistochemistry reagents and kits.
(a)
Identification. Immunohistochemistry test systems (IHC's) are in vitro diagnostic devices consisting of polyclonal or monoclonal antibodies labeled with directions for use and performance claims, which may be packaged with ancillary reagents in kits. Their intended use is to identify, by immunological techniques, antigens in tissues or cytologic specimens. Similar devices intended for use with flow cytometry devices are not considered IHC's.(b)
Classification of immunohistochemistry devices. (1) Class I (general controls). Except as described in paragraphs (b)(2) and (b)(3) of this section, these devices are exempt from the premarket notification requirements in part 807, subpart E of this chapter. This exemption applies to IHC's that provide the pathologist with adjunctive diagnostic information that may be incorporated into the pathologist's report, but that is not ordinarily reported to the clinician as an independent finding. These IHC's are used after the primary diagnosis of tumor (neoplasm) has been made by conventional histopathology using nonimmunologic histochemical stains, such as hematoxylin and eosin. Examples of class I IHC's are differentiation markers that are used as adjunctive tests to subclassify tumors, such as keratin.(2) Class II (special control, guidance document: “FDA Guidance for Submission of Immunohistochemistry Applications to the FDA,” Center for Devices and Radiologic Health, 1998). These IHC's are intended for the detection and/or measurement of certain target analytes in order to provide prognostic or predictive data that are not directly confirmed by routine histopathologic internal and external control specimens. These IHC's provide the pathologist with information that is ordinarily reported as independent diagnostic information to the ordering clinician, and the claims associated with these data are widely accepted and supported by valid scientific evidence. Examples of class II IHC's are those intended for semiquantitative measurement of an analyte, such as hormone receptors in breast cancer.
(3) Class III (premarket approval). IHC's intended for any use not described in paragraphs (b)(1) or (b)(2) of this section.
(c)
Date of PMA or notice of completion of a PDP is required. As of May 28, 1976, an approval under section 515 of the Federal Food, Drug, and Cosmetic Act is required for any device described in paragraph (b)(3) of this section before this device may be commercially distributed. See § 864.3.
0
August 15, 2022
Image /page/0/Picture/1 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: on the left, there is a seal with an eagle emblem, and on the right, there is the text "FDA U.S. FOOD & DRUG ADMINISTRATION" in blue. The text is arranged in three lines, with "FDA" in a larger font size and a blue square behind it.
Boston Cell Standards, Inc. % Gail Radcliffe, President Radcliffe Consulting, Inc. 23 Fairbanks St West Boylston, MA 01583
Re: K220163
Trade/Device Name: HER2/ER/PR IHControls® - Level H HER2/ER/PR IHControls® - Level M HER2/ER/PR IHControls® - Level L Regulation Number: 21 CFR 864.1860 Regulation Name: Immunohistochemistry Reagents And Kits Regulatory Class: Class II Product Code: NJW Dated: January 10, 2022 Received: January 20, 2022
Dear Gail Radcliffe:
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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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
1
requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Shyam Kalavar Deputy Branch Chief Division of Molecular Genetics and Pathology OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
2
Indications for Use
510(k) Number (if known) K220163
Device Name Her2/ER/PR IHControls® - Level H Her2/ER/PR IHControls® - Level M Her2/ER/PR IHControls® - Level L
Indications for Use (Describe)
HER2/ER/PR IHControls® -Level H
HER2/ER/PR IHControls® -Level H are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast turnor samples.
HER2/ER/PR IHControls® -Level H are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level H are an additional controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® -Level M
HER2/ER/PR IHControls® -Level M are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast turnor samples.
HER2/ER/PR IHControls® -Level M are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level M are an additional controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® -Level L
HER2/ER/PR IHControls® -Level L are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® -Level L are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level L are an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
3
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4
5. 510(k) Summary
5.1 Boston Cell Standards (BCS)
Sponsor:
Boston Cell Standards Inc. 800 Washington St. Boston, MA 02111 Tel: 617-636-8042
Contact Person:
Steven Bogen, MD, PhD Boston Cell Standards Inc. 800 Washington St. Boston MA 02111 E-mail: sbogen@bostoncellstandards.com Tel: 617-636-5422 FAX: 617-674-3570
Consultant:
Gail Radcliffe, PhD Radcliffe Consulting, Inc. 231 Fairbanks St. West Boylston, MA 01583 E-mail: gradcliffe@RMDCI.com Tel: 508-934-9673
5.2 Device
Trade Name: HER2/ER/PR IHControls® - Level H HER2/ER/PR IHControls® - Level M HER2/ER/PR IHControls® - Level L Common name: Control Material, peptide-based controls for HER2, ER, and PR, Immunohistochemistry Regulation number: 864.1860 Product Code: NJW Class: 2
5.3 Predicate Device
QCS HER2 Immunocontrols (K023335)
5
5.4 Intended Use
HER2/ER/PR IHControls® - Level H
HER2/ER/PR IHControls® - Level H are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® - Level H are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® - Level H are an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® - Level M
HER2/ER/PR IHControls® - Level M are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® - Level M are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® - Level Mare an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® - Level L
HER2/ER/PR IHControls® - Level L are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® - Level L are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® - Level L are an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
6
5.5 Device Description
HER2/ER/PR IHControls® - Level H, HER2/ER/PR IHControls® - Level M and HER2/ER/PR IHControls® - Level L (HER2/ER/PR IHControls®) are immunostaining positive controls for diagnostic immunohistochemistry laboratories. The same immunohistochemical reaction that occurs on a tissue or cellular sample also occurs on the HER2/ER/PR IHControls® microbead. As a result of immunostaining, the microbead bearing the analyte turns the same color as cells expressing the analyte.
HER2/ER/PR IHControls® incorporate assay controls for human epidermal growth factor receptor type II (HER-2), estrogen receptor (ER), and progesterone receptor (PR). The HER2/ER/PR panel includes controls at 3 different analyte concentrations, providing users with an opportunity to select the control with an analyte concentration within the dynamic range of their assay.
HER2/ER/PR IHControls® are provided in a small vial containing enough liquid suspension for at least 100 tests. The user is instructed to pipet a one microliter droplet onto the slide that also bears the patient sample. The droplet is a suspension containing microscopic glass microbeads that are approximately the same size as cells. The microbead surface bears the molecule(s) being measured. Peptides representing the epitopes of all of the major commercial HER2. ER. and PR antibodies used by clinical immunohistochemistry laboratories are incorporated into the products.
HER2/ER/PR IHControls® are comprised of two different microbeads: analyte-coated glass test microbeads (7-8 um in diameter) and color standard microbeads (4.5 um in diameter). The latter provide a fixed brown color intensity for use in standardizing stain intensity measurement.
The HER2/ER/PR IHControls® panel is provided at 3 different analyte concentrations. In descending order of concentration, these levels are denoted "H", "M", and "L". These different products allow matching the analyte concentration to the dynamic range of the laboratory's stain.
| Level H (high) | This product includes analytes for HER2, ER, and PR, at approximately
$10^6$ molecules per microbead. |
|------------------|----------------------------------------------------------------------------------------------------------|
| Level M (medium) | This product includes analytes for HER2, ER, and PR, at approximately
$10^5$ molecules per microbead. |
| Level L (low) | This product includes analytes for HER2, ER, and PR, at approximately
$10^4$ molecules per microbead. |
These concentrations are approximate; they are not intended as assayed controls.
Three product levels are provided to allow the user to select a control with the lowest analyte concentration that still produces an easily visible immunostain.
The immunoreactivity pattern for product levels H, M, and L is described in Table 1. The table indicates the primary antibodies corresponding with up to 9 separate HER2/ER/PR peptides.
7
(Some peptides are immunoreactive with more than one primary antibody.) Level H has a narrower range of primary antibody immunoreactivity than levels M and L but the concentrations per microbead are higher. Although any individual primary antibody will be listed as potentially immunoreactive with 2 or more levels. IHC laboratories should use the lowest level that provides for an easily visible stain.
| Level H | Level M | Level L |
|---|---|---|
| (BRLS11) | (BRL2U04) | (BRL2W08) |
| HER2 CB11 | HER2 CB11 | HER2 CB11 |
| HER2 4B5 | HER2 4B5 | HER2 4B5 |
| HercepTest | HercepTest | HercepTest |
| PR 636 | PR 636 | PR 636 |
| PR 16 | PR 16 | PR 16 |
| ER EP1 | ER EP1 | ER EP1 |
| ER 1D5/2.123 | ER 1D5/2.123 | |
| ER 6F11 | ER 6F11 | |
| PR 1294 | PR 1294 | |
| PR 1E2 | PR 1E2 | |
| ER SP1 | ER SP1 |
| Table 1. HER2/ER/PR IHControls® Immunoreactivity: Levels H. M. and L | ||||||
|---|---|---|---|---|---|---|
| ---------------------------------------------------------------------- | -- | -- | -- | -- | -- | -- |
Users pipette a droplet onto a microscope slide that also bears a patient's tissue sample. The droplet hardens upon drying, adheres the analyte-coated microbeads to the microscope slide, and is then processed with the patient sample. The hardened droplet is able to pass through dry heat associated with "baking" of slides, organic solvents associated with deparaffinization, boiling associated with antigen retrieval, and repeated rinses associated with immunostaining.
5.6 Principle of Operation
The HER2/ER/PR IHControls® contain analytes that are immunoreactive in IHC stains. They are present on every slide. Therefore, negative staining due to analytic error is readily detectable. Like tissue samples, the HER2/ER/PR IHControls® are formalin-fixed during manufacture. Therefore, test microbeads in the HER2/ER/PR IHControls® products may demonstrate improved immunoreactivity after antigen retrieval.
5.7 Substantial Equivalence
The similarities and differences between Boston Cell Standards HER2/ER/PR IHControls® and the predicate (QCS control slides for HER2 IHC) are outlined in the table below.
8
Table 2. Comparison of HER2/ER/PR IHControls® Panel to Predicate
9
| Control
configuration | not intended to be used for scoring
HER2, ER, and PR IHC stained slides.
Her2/ER/PR IHControls® -Level M are
an additional control to the run controls
specified in the HER2, ER, or PR IHC
device labeling and are not intended to
replace the controls approved or cleared
as part of an IHC device.
Her2/ER/PR IHControls® -Level L
Her2/ER/PR IHControls® -Level L are
peptide based qualitative on-slide
controls to monitor the performance of
the analytic components (antigen retrieval
and immunostaining) of the
immunohistochemical (IHC) staining
process for certain human epidermal
growth factor receptor type II (HER2),
estrogen receptor (ER) and progesterone
receptor (PR) IHC stains. It is indicated
for use with formalin-fixed paraffin-
embedded (FFPE) breast tumor samples.
Her2/ER/PR IHControls® -Level L are
not intended to be used for scoring
HER2, ER, and PR IHC stained slides.
Her2/ER/PR IHControls® -Level L are
an additional control to the run controls
specified in the HER2, ER, or PR IHC
device labeling and are not intended to
replace the controls approved or cleared
as part of an IHC device. | Breast cancer cell lines (MDA-
361, MDA-453, MCF-7) for
detection of HER2. |
|--------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------|
| | Level H
This product includes analytes for
HER2, ER, and PR, at approximately
106 molecules per microbead. | |
| | Level M
This product includes analytes for
HER2, ER, and PR, at approximately
105 molecules per microbead. | |
| | Level L
This product includes analytes for | |
10
| | HER2, ER, and PR, at approximately
104 molecules per microbead. | |
|--------------------------|------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------|
| Assay
compatibility | Polyclonal and monoclonal IHC stains | Polyclonal and monoclonal IHC
stains |
| Analyte
concentration | Three different levels of HER2,
ER, PR each at different
concentration. | Three different cell lines each
expressing unknown
concentrations of HER2. |
| | Differences | |
| Reagents | Formalin-fixed HER2, ER, PR peptide
epitopes covalently attached to glass
microbeads in a liquid matrix that
adhere to a slide. | Formalin-fixed paraffin
embedded breast cancer cell
lines expressing HER2
protein, mounted on slides. |
| Slide
controls | Controls on every slide, as per
regulatory guidelines. | Separate slides for batch
control |
5.8 Analytical Performance Studies
5.8.1 Specificity
The HER2/ER/PR IHControls® were tested against twenty-six (26) antigenically irrelevant, commonly used primary antibodies, in the context of clinical IHC testing (Table 3). Positive tissue controls demonstrated that the antigenically irrelevant tests were operating correctly. No cross-reactivity was detected with the HER2/ER/PR IHControls®. When an appropriate ER, HER2 and PR primary antibody was used for staining, HER2/ER/PR IHControls® demonstrated immunoreactivity.
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| | Marker | Organ | Antibody | Tissue
Control | IH Controls® |
|-----|-------------------|-------------|---------------|-------------------|--------------|
| | | | | | |
| 1. | Bcl-6 | Tonsil | GI191E/A8 | Positive | Negative |
| 2. | C4d | Tonsil | SP91 | Positive | Negative |
| 3. | CD10 | Tonsil | SP67 | Positive | Negative |
| 4. | CD138 | Tonsil | B-A38 | Positive | Negative |
| 5. | CD20 | Tonsil | L26 | Positive | Negative |
| 6. | CD34 | Tonsil | QBEnd/10 | Positive | Negative |
| 7. | CD5 | Tonsil | SP19 | Positive | Negative |
| 8. | CD56 | Tonsil | MRQ-42 | Positive | Negative |
| 9. | CMV | Placenta | CMV | Positive | Negative |
| 10. | Cyclin D1 | Lymphoma | SP4-R | Positive | Negative |
| 11. | Cytokeratin
20 | Colon | SP33 | Positive | Negative |
| 12. | Cytokeratin 7 | Lung | SP52 | Positive | Negative |
| 13. | EFGR | Normal Skin | 5B7 | Positive | Negative |
| 14. | Keratin | Skin | 34BE12 | Positive | Negative |
| 15. | Ki-67 | Tonsil | 30-9 | Positive | Negative |
| 16. | Mart-1 | Melanoma | A103 | Positive | Negative |
| 17. | MITF | Melanoma | C5/D5 | Positive | Negative |
| 18. | p53 | p53 | BP53-11 | Positive | Negative |
| 19. | p63 | Prostate | 4A4 | Positive | Negative |
| 20. | Pan Keratin | Appendix | AE1/AE3/PCK26 | Positive | Negative |
| 21. | Podoplanin | Tonsil | D2-40 | Positive | Negative |
| 22. | PSA | Prostate | PSA | Positive | Negative |
| 23. | S100 | S100 | 4C4.9 | Positive | Negative |
| 24. | Somatostatin | Pancreas | Somatostatin | Positive | Negative |
| 25. | TAG 72 | Colon | B72.3 | Positive | Negative |
| 26. | TTF-1 | Thyroid | 8G7G3/1 | Positive | Negative |
| 27. | Breast | Breast ca | ER SP1 | Positive | Positive |
| 28. | Breast | Breast ca | HER2 4B5 | Positive | Positive |
| 29. | Breast | Breast ca | PR 1F2 | Positive | Positive |
Table 3. Specificity Testing - List of Primary Antibodies, Targets, and Test Results
5.8.2 Shelf Life
The shelf life for each HER2/ER/PR IHControl® level, when stored at 2 - 8 ° C, was tested in real time over approximately 2 years. Table 4 summarizes product shelf life.
| Table 4. Product Shelf-Life | |
|---|---|
| Product | Shelf Life (days) |
| Level H (BRLS11) | 392 |
| Level M (BRL2U04) | 392 |
| Level L (BRL2W08) | 392 |
5.8.3 Reproducibility of Pathologist IHControl® Readouts
Three pathologists evaluated a blinded slide set of 30 stained HER2/ER/PR IHControl® slides.
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Ten (10) slides each were stained for HER2, ER, and PR. In each set of ten, 4 were stained with a diluted primary antibody, or no primary antibody, to create weak or absent staining. Overall agreement (Table 5) among pathologists for the interpretation of HER2/ER/PR IHControls® was 100%. Pathologists are able to visually interpret HER2/ER/PR IHControls® and reproducibly render Pass or Fail scores.
| Slide number | Pathologist 1 | Pathologist 2 | Pathologist 3 | Expected Result | Concordance |
|---|---|---|---|---|---|
| HER2 | |||||
| 1 | P | P | P | P | 100% |
| 2 | P | P | P | P | 100% |
| 3 | F | F | F | F | 100% |
| 4 | P | P | P | P | 100% |
| 5 | F | F | F | F | 100% |
| 6 | P | P | P | P | 100% |
| 7 | F | F | F | F | 100% |
| 8 | P | P | P | P | 100% |
| 9 | F | F | F | F | 100% |
| 10 | P | P | P | P | 100% |
| ER | |||||
| 11 | P | P | P | P | 100% |
| 12 | P | P | P | P | 100% |
| 13 | F | F | F | F | 100% |
| 14 | P | P | P | P | 100% |
| 15 | F | F | F | F | 100% |
| 16 | P | P | P | P | 100% |
| 17 | F | F | F | F | 100% |
| 18 | P | P | P | P | 100% |
| 19 | F | F | F | F | 100% |
| 20 | P | P | P | P | 100% |
| PR | |||||
| 21 | P | P | P | P | 100% |
| 22 | P | P | P | P | 100% |
| 23 | F | F | F | F | 100% |
| 24 | P | P | P | P | 100% |
| 25 | F | F | F | F | 100% |
| 26 | P | P | P | P | 100% |
| 27 | F | F | F | F | 100% |
| 28 | P | P | P | P | 100% |
| 29 | F | F | F | F | 100% |
| 30 | P | P | P | P | 100% |
Table 5. Pathologist Reader Reproducibility
5.9 Clinical Performance Studies
The HER2/ER/PR IHControls® were incorporated as on-slide controls for HER2, ER, and/or PR immunostaining at three clinical immunohistochemical laboratories. The laboratories incorporated the HER2/ER/PR IHControls® but otherwise followed their typical method for patient testing. After immunostaining was complete, the laboratories performed an assessment. For each stained slide, the pathologist interpreted both the conventional (tissue) control and the HER2/ER/PR IHControls®, for each of the HER2, ER, and PR stains. The controls (tissue
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controls or HER2/ER/PR IHControls®) were scored as either "Pass" or "Fail", depending on whether the control shows that the test passed or failed the quality control check. Percent concordance is reported.
The overall concordance rate is 440 out of 442 tests, or 99.5%, from all 3 clinical trial sites, for ER, PR, and HER2 tests combined (Table 6). Individual site concordance was 100%, 99.5%, and 99.1%.
Two discrepancies between tissue controls and HER2/ER/PR IHControls® were due to operator error: (1) use of an incorrect HER2/ER/PR IHControl®, and (2) placement of the HER2/ER/PR IHControls® at the slide edge because the patient sample and tissue (comparator) control left no other place. There was also one site where a histotechnologist did not pipette the HER2/ER/PR IHControls® onto the slide.
| Site | Immunostain | Product Tested | # Slides | Concurrence | Deviations |
|---|---|---|---|---|---|
| 1 | ER NCL-L-ER-6F11 | BRL2U04-001 | 39 | 39/39 | 0 |
| 1 | PR NCL-L-PGR-312 | BRLS11-001 | 39 | 39/39 | 0 |
| 1 | HER2 BOND Oracle | BRL2U04-001 | 39 | 39/39 | 0 |
| 2 | PR Agilent M3569 | BRL2U04-001 | 172* | 171/172 | 1 |
| 2 | HER2 PATHWAY | BRLS11-003 | 42 | 42/42 | 0 |
| 3 | ER CONFIRM | BRL2W08-003 | 37 | 36/37 | 1 |
| 3 | PR BOND PA0312 | BRLS11-001 | 37 | 37/37 | 0 |
| 3 | HER2 PATHWAY | BRLS11-001 | 37 | 37/37 | 0 |
Table 6. Data Summary from Five Clinical Sites
- Slides from two days were deemed uninformative and not counts for the difference between 182 slides tested and the denominator in the concurrence column being 172, as there were 5 slides per day that did not generate any data.
The data (Table 6) demonstrate that the HER2/ER/PR IHControls® provide an equally accurate assessment of IHC test quality.
6.0 Conclusions from Nonclinical and Clinical Data
The conclusions drawn from the analytical and clinical data demonstrate that the device is safe and effective for its intended use.