Monoclonal Mouse Anti-Human Estrogen Receptor, Clone 1D5 may be used in the semi-quantitative detection of human estrogen receptor in tissue sections of human breast cancer by immunohistochemistry. The information gained by this assay can aid in assessing the likelihood of response to therapy as well as in the prognosis and management of breast cancer patients.

The clinical interpretation of any positive staining or its absence should be complemented by morphological and histological studies with proper controls. Evaluations should be made within the context of the patient's clinical history and other diagnostic tests by a qualified individual having knowledge of all the potential antibody reactivities.

Device Description

Monoclonal Mouse Anti-Human Estrogen Receptor, Clone 1D5, Device Code No. M7047) is a mouse anti-human monoclonal antibody (Product) produced as a tissue culture supernatant. The antibody is supplied in Description: 0.05M Tris-HCl buffer, pH 7.2, containing fotal bovine serum and 15mM sodium azide. (1mL total volume).
Monoclonal Mouse Anti-Human Estrogen Receptor. Clone 105 Ready-to-Use Antibody and Negative Control (Product Code No. N1576) consists of a mouse anti-human monoclonal antibody produced as a tissue culture supernatant and pre-diluted in 0.05M Tris-HCI buffer, pH 7.6, containing fotal bovine serum and 15mM sodium azide (7mL total volume). The primary antibody is packaged with a negative control reagent consisting of a cocktail of purified mouse immunoglobulins (IgG, IgG, IgGz, IgGz, IgGz and IgM) in 0.05M Tris-HCI buffer, pH 7.6 and 15mM sodium azide (5mL total volume).

AI/ML Overview

The provided text describes DAKO's Monoclonal Mouse Anti-Human Estrogen Receptor, Clone 1D5 (anti-ER, 1D5) and its comparison to a predicate device, Abbott ER-ICA Monoclonal, clone H222. The studies performed focused on demonstrating substantial equivalence, rather than establishing specific quantitative acceptance criteria for the anti-ER, 1D5 device alone against predefined thresholds.

However, based on the comparative effectiveness studies presented, we can infer performance and the methods used to demonstrate the device's utility.

Here's an analysis based on the provided text:

1. Table of Acceptance Criteria and Reported Device Performance

The submission does not explicitly state quantitative "acceptance criteria" for the anti-ER, 1D5 antibody against a specific clinical outcome. Instead, it demonstrates performance by comparing it to an already approved predicate device (Abbott ER-ICA H222) and by citing published studies on its use. The primary goal was to establish substantial equivalence.

The table below summarizes the reported performance of anti-ER, 1D5 and its comparison to the predicate device and clinical outcomes, where available.

Metric (Inferred Acceptance Criterion based on predicate comparison or clinical relevance)	Reported Device Performance (anti-ER, 1D5)
Normal Tissue Reactivity (Expected specific staining, no non-specific staining)	Expected positive staining in breast, cervix, and uterus. No reactivity in ER-negative tissues. Weak cytoplasmic staining considered non-specific.
Reproducibility (Consistent results within and between runs)	Consistent results with intra- and inter-run testing.
Specificity vs. Predicate (H222)	Varied from 51% to 79% (across different studies). One study reported 79.2% specificity. Another reported 88% specificity with H222 frozen and 64.90% with H222 paraffin.
Sensitivity vs. Predicate (H222)	Varied from 89% to 100% (across different studies). One study reported 94.6% sensitivity. Another reported 91.95% sensitivity with H222 paraffin and 100% with H222 frozen.
Positive Predictive Value vs. Predicate (H222)	One study reported 64%.
Negative Predictive Value vs. Predicate (H222)	One study reported 92%.
Correlation with Outcome (Tamoxifen Response)	Predictive information for selecting patients benefiting from hormonal treatment. Strong predictor of favorable primary response in 89% of 72 cases in one study.
Correlation with Overall Survival (OS)	Significant for DFS (p = 0.01) and OS (p = 0.01) in one study (for 3+ intensity).
Correlation with Relapse-Free Survival (RFS)	Prognostically relevant for predicting relapse-free survival in node-positive cases.
Correlation with H222 frozen (R value, if reported)	R=0.7, with p<0.0001 (in one study comparing H scores).

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

The "test set" here refers to the various studies cited in the submission for comparative effectiveness and clinical utility. There isn't a single "test set" but rather a collection of published research:

Normal Tissue Testing: A "required panel of normal tissues" as specified by FDA guidance. The exact number is not explicitly stated in the summary but indicates a prospective testing approach.
Reproducibility Testing: Eight serial sections from each of three different paraffin-embedded blocks of human breast carcinoma. This was an internal prospective study.
Comparative Effectiveness and Clinical Utility Studies (Table 1): This table summarizes findings from various published articles.
- Mauri, et al.: 374 samples.
- Goulding, et al.: 90 samples.
- Nedergaard et al.: 83 samples.
- Pertschuk et al.: 74 samples (all had Stage IV disease; included White, AfroAm, Hispanic, Asian demographics).
- Leong and Milos.: 31 samples.
- Pellicer and Sundblad: 300 patients with 5-year follow-up.
- Hopkins et al.: 51 samples.
- Hendricks and Wilkinson: 20 samples.

Data Provenance for Published Studies: The text does not explicitly state the country of origin for each study, but given the journal names and author affiliations typically found in such publications, it's highly likely to be a mix of international (including US and European) retrospective cohort studies. The studies are consistently referred to as "Published Immunoreactivity."

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

The text does not specify the number or qualifications of experts used to establish ground truth for the test set utilized in the submission. The studies cited (Table 1) often compare the anti-ER, 1D5 results to the predicate device (H222) or to clinical outcomes like response to tamoxifen.

In the case of comparison to H222 (predicate device), the ground truth for H222 results would have been established by trained pathologists.
For clinical outcome data (e.g., tamoxifen response, survival), the ground truth is derived from patient medical records and follow-up data, generally managed and interpreted by clinicians.
The scoring systems mentioned (e.g., percentage of stained tumor nuclei, H scores, 0-3+ intensity) imply evaluation by trained individuals, likely pathologists, but their number and specific qualifications are not detailed in this summary.

4. Adjudication Method for the Test Set

The submission does not explicitly describe an adjudication method (like 2+1 or 3+1) for the test set examples presented. The performance metrics are reported from individual published studies, which would have had their own internal methods for slide interpretation and data analysis. These methods often involve interpretation by one or more pathologists, but explicit adjudication rules are not provided in this summary.

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

No, a Multi Reader Multi Case (MRMC) comparative effectiveness study was not done involving AI assistance. This submission describes an antibody for immunohistochemical staining, not an AI-powered diagnostic device. The comparisons are between two different antibodies (DAKO's 1D5 and Abbott's H222) and their correlation with clinical outcomes.

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

No, a standalone (algorithm-only) performance study was not conducted. This is not an AI device. The device is a diagnostic reagent (antibody) that is interpreted by human observers (pathologists) under a light microscope. The text mentions "image analysis systems" and "automated stainers" as being used in efforts for greater objectivity but notes they "have yet to be validated in the literature," implying they are not part of the core performance claims for this specific 510(k) submission.

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.)

The ground truth used across the various studies cited is a combination of:

Pathology: Evaluation of tissue sections by pathologists for the presence and quantity of estrogen receptor staining (both for the predicate device H222 and for anti-ER, 1D5). This includes semiquantitative scoring systems (e.g., percentage of positive cells, staining intensity, H scores).
Clinical Outcomes Data: Patient follow-up data related to response to therapy (e.g., tamoxifen), overall survival, and relapse-free survival.
Expert Consensus (Implicit): While not explicitly stated as a formal "expert consensus" process for establishing ground truth within the context of the submission's own testing, the predicate device (Abbott ER-ICA H222) had previously been approved by the FDA based on its established utility and, presumably, expert agreement on its diagnostic accuracy. The studies comparing anti-ER, 1D5 to H222 are effectively using H222 results (interpreted by pathologists) as a reference.

8. The Sample Size for the Training Set

The concept of a "training set" is not applicable in the context of this device, as it is not an AI/machine learning algorithm. The studies described are for validation and comparison of a biological reagent (antibody).

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

As this is not an AI/machine learning device, there is no "training set" for which ground truth would be established. The information gathered about the antibody's characteristics and performance came from its development, internal testing, and independent published research.

Summary

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MAR - 3 2000

510{k) Summary

Submitter:	DAKO Corporation6392 Via RealCarpinteria, CA 93013805-566-6655
Contact:	Gretchen M. Murray, Ph.D., Regulatory Affairs Manager
Date SummaryPrepared:	October 30, 1999 (amended March 2, 2000)
Device Name:	1) DAKO® Monoclonal Mouse Anti-Human Estrogen Receptor, Clone1D5, Antibody for Immunoenzymatic Staining (Product Code No.M7047)2) DAKO® Monoclonal Mouse Anti-Human Estrogen Receptor, Clone1D5, Ready-to-Use Antibody and Negative Control Reagent forImmunoenzymatic Staining (Product Code No.N1575)
DeviceClassification:	Class II for prognostic immunohistochemical staining reagents (21CFR 864.1860).
Panel:	Hematology and Pathology Devices Panel.Division of Clinical Laboratory Devices.
Predicate Device:	Abbott ER-ICA Monoclonal, Clone H222 approved by the FDA as PM# P880026 and downclassified to class II by 21 CFR 864.1860,Immunohistochemistry Reagents and Kits on June 3, 1998. Devicepackage insert from this product is included in Section 2 of thissubmission.

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Monocional Mouse Anti-Human Estrogen Receptor, Clone 1D5, Device Code No. M7047) is a mouse anti-human monoclonal antibody (Product) produced as a tissue culture supernatant. The antibody is supplied in Description: 0.05M Tris-HCl buffer, pH 7.2, containing fotal bovine serum and 15mM sodium azide. (1mL total volume).

Monoclonal Mouse Anti-Human Estrogen Receptor. Clone 105 Ready-to-Use Antibody and Negative Control (Product Code No. N1576) consists of a mouse anti-human monoclonal antibody produced as a tissue culture supernatant and pre-diluted in 0.05M Tris-HCI buffer, pH 7.6, containing fotal bovine serum and 15mM sodium azide (7mL total volume). The primary antibody is packaged with a negative control reagent consisting of a cocktail of purified mouse immunoglobulins (IgG, IgG, IgGz, IgGz, IgGz and IgM) in 0.05M Tris-HCI buffer, pH 7.6 and 15mM sodium azide (5mL total volume).

Intended Use: For In Vitro Diagnostic Use

Monoclonal Mouse Anti-Human Estrogen Receptor, Clone 1D5 may be used in the semiquantitative detection of human estrogen receptor in tissue sections of human breast cancer by immunohistochemistry. The information gained by this assay can aid in assessing the likelihood of respanse to therapy as well as in the prognosis and management of breast cancer patients.

Statement of Substantial Equivalence:

DAKO" Monoclonal Mouse Anti-Human Estrogen Receptor, 1D5 (anti-ER, 1D5) is a monoclonal antibody produced as a tissue culture supernatant which is available as a concentrate (Code No. M7047) and a prediluted primary (Code No. N1675),

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Anti-ER. 1D5 immunohistochemically demonstrates the presence of estrogen receptor in a variety of normal and pathological tissues. In normal tissue, anti-ER, 1D5 labels estrogen containing cells. Pathological reactivity includes detection of estrogen receptor in breast cancer.

Anti-ER, 1D6 is comparable in use and technology to Abbott ER-ICA Monoclonal, clone H222 which labels cells in tissues and is currently in commercial distribution. Similarities of the anti-ER, 1D5 to the Abbott ER-ICA include that both products are monoclonal antibodies directed against the human Estrogen Receptor (ER). The differences between the two products include that the DAKO anti-ER, 1D5 is directed to the N-terminal end of the ER, while the H222 clone from the Abbott ER-ICA is directed against the functional section of the ER, coded for in exon 5 of the ER DNA. Clone H222 is of rat origin, while anti-ER, 1D5 is of mouse origin. Functionally, anti-ER, 1D5 reacts with an epitope exposed in formalin-fixed, paraffin-embedded tissues by heat induced epitope retrieval. The H222 clone binds to ER found in frozen tissues. When the H222 clone has been tested on formalin-fixed, paraffin-embedded tissues, the specificity was decreased from that seen with frozen tissues.

Visualization of the primary antibody for the Abbott ER-ICA is achieved using an indirect peroxidase anti-peroxidase (PAP) system. ER-ICA monoclonal is located by a goat anti-rat bridging antibody. Anti-Rat PAP complex is added, which recognizes the bridging antibody. Then a precipitating enzyme generated product (hydrogen peroxide) and a chromogan substrate solution, containing DAB are added. The reaction product produces a reddish brown precipitate at the locations of the primary antibody.

The DAKO anti-estrogen receptor, clone 1D5 is substantially equivalent to the Abbott ER-ICA Monoclonal, clone H222 in that both products specifically bind to estrogen receptor proteins located in the nuclei of cells. Bath products require similar detection chemistry principles for visualization of the product, and both aid in the prognosis of breast carcinoma.

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Functional comparison testing of anti-ER, 1D5 on formalin-fixed paraffin-ambedded specimens and Abbort's ER-ICA using frozen breast cancer specimens is presented in Table 1.

Experimental Data 1.

Normal Tissue Testing:

(The negative raagent control for these 2 studies was fetal bovine serum diluted with the same Tris-HCl buffer with 0.015M NaN3 as the primary antibody.)

The required panel of normal tissues was tested with this antibody as specified in the 6/3/98 final version of Guidance for Submissions of Immunohistochemistry Applications to the FDA. All tissues were formalin fixed and paraffin embedded. Staining was performed using the DAKO LSAB®2 Peroxidase kit system (Code No. K0672).

Normal tissues exhibiting positive staining with anti-ER, 1D5 included the following: breast, cervix and uterus. Refer to the package insert Table 2, Normal Tissue Reactivity for additional testing results or to Section 3 of this submission for the Results of the Normal Tissue Reactivity Testing.

Reproducibility Testing:

Eight serial sections from each of three different paraffin embedded blocks of human breast carcinoma were collected for testing. Testing was performed as follows:

Intra-run reproducibility: Following the standard DAKO LSAB®2 Peroxidase Kit protocol (Code No. K0677), three slides from each tissue block were steined with Monoclonal Mouse Anti-Human Estrogen Receptor, Clone 1D5 Ready-to-Use Antibody and negative control reagent consisting of fatal bovine serum in the equivalent diluent. Concurrently, one slide from each block was stained with the supplied negative control reagent.

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Inter-run reproducibility: Staining one slide from each tissue block, the above procedure was repeated on two additional days. Concurrently, one slide from each block was stained with the supplied negative control reagent.

Reproducibility experiments with anti-ER. 1DE yielded consistent results with intra- and inter-run testing. Consistent test conditions were maintained throughout the study and reagents were stored at 2-8° C between test runs. (See Section 3 for the Report of the Results of the Reproducibility Testing.)

)). Published Immunoreactivity

Twenty-six articles published on the characterization or clinical use of estrogen receptor were used in the submission. Twenty-three of those articles reported on studies using anti-ER, 1D5. Following is a brief summary of the compiled information.

Estrogen Receptor Characteristics

Steroid receptors exhibit a high affinity and specificity for their ligands. The human estrogen receptor (ER) is a dimeric protein of 65 kDa located primarily on the membrane of cell nuclei and belongs to a class of trans-acting proteins which stimulate transcription by binding to specific DNA elements, also known as hormone response elements. Through binding estrogen, the ER is induced to precisely stimulate gene transcription, hence is also known as an inducible enhancer factor. 1

Measurement of the ER has been shown to be important in the initial evaluation of breast cancer patients, for providing information on the likelihood of a successful endocrine response and for the management of the patiant and the tumor. 2.3

Development of ER-1D5 monoclonal antibody

The monoclonal antibody, anti-ER, 1D6 was produced in BALB/c mice by i.p. injection of recombinant estrogen receptor (RER). * The RER used was obtained by insarting human ER

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cDNA into the Ndel-BamHI site of translation vector pETIla and introducing the same into the 5' end of the coding sequence of ER, a fragment of the plasmid pSG5-HGO. This modified cDNA was then inserted into the plasmid expression vector pETIla. E. coli, transformed with pETII-HGO, and produced a protein with molecular mass of 67kD. Clone 1D5 was shown to react with an epitope located in the N-terminal domain of ER.

Clone 1D5 Positivity in Normal Tissues

In normal frozen and formalin-fixed, paraffin-embedded human tissue, anti-ER, 1D5 reacts with the nuclei of cells known to contain large amounts of ER, including cells of the mammary gland and the uterus. No reactivity was found in tissues normally considered negative for ER. 4 When present, weak cytoplasmic staining was considered nonspecific.

Clone 1D5 Presence in Abnormal Tissues

In pathological tissues, anti-ER, 1D5 was examined for specificity and sensitivity for breast cancer as well as for the evaluation of patients for endocrine therapy. Numerous studies of cases of breast cancer showed anti-ER, 1D5 to be safe and effective in this endoavor. 513 On frozen tissues, anti-ER, 1D6 immunoreacted with 63/93 (67.7%) cases of broast cancer and only 1/30 (3.3%) nonbreast cancers reacted positively. 4 Direct comparison of anti-ER, 1D5 on formalin-fixed, paraffin-embodded (FFPE) specimens with H222 on frozen sections of the same tumors has been completed by 7 different laboratories on more than 1000 specimens. The results are reported in Table 1.

The specificity of anti-ER, 1D5 (aither assessed as compared to H222 or to outcome with tamoxifon) varied from 51% to 79% while sensitivity varied from 89% to 100%. H222 was reported to be less sensitive but more specific in one article. "* Thus, the use of IHC with anti-ER, 1D5 correlates well with a previously approved immunocytochemical assay for ER. However, clinical follow-uns have chown that IHC hu use of anti-FR 105 nrovided

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and and the contribution of the first to the program and comments of the

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Definition of positive IHC assay results with anti-ER. 1D5 have ranged somewhat arbitrarily from a minimum of any calls staining positive, to 5% of stained tumor nuclei, 21 to 10% " 11.13.15.18 22 or 20%.18 In over 900 cases of breast cancer, one investigator observed a strong relationship between staining intensity and the number of positive cells (g 0.001).ª Because of the above arbitrary limits of positivity, these authors and others 4, 20, 22-28 designed a semiguantitative "score" from the percentage of stained cells and the nuclear staining intensity (e.g. 0-3 or negative, weak, moderate, and strong). This scoring system is adopted for anti-ER, 1D5. Howaver, Pertchuck et al (1996) found little support for the use of such scores. 13

lmage analysis systems 7,8,19 and automated stainers 5 were used in an effort to obtain greater objectivity in assessing positive expressions but have yet to be validated in the literature.

In 105 cases of node-negative primary breast cancer, anti-ER, 105-positivity made no significant difference in predicting overall survival, whereas in 152 node-positive casas, anti-ER, 1D5-positivity was found to be prognostically relevant for predicting overall survival and predicting relapse-free survival.11 Staining with anti-ER, 106 provided predictive information for the selection of patients who may benefit by hormonal treatment. 1 Image analysis of stained tissues from 250 cases of breast cancer showed disease progression to be 1.7 times and death rate to be 2.5 times higher in anti-ER, 1D5negative cases than in 1D5-positive cases if the cut-off value was "optimally" salected. 4 Of 36 Anti-ER, 1 D5-positive cases, 23 (64%) showed favorable responses to therapy, whereas 35/38 (92%) 1D5-negative cases showed disease progression. 13 Others 19 have reported that 1D5-positivity was a strong prodictor of favorable primary response to tamoxifen in 89% of 72 cases and that treatment did not impede the detection of ER by anti-ER, 1D5.

Bibliography:

1. Kurnar V, Green S, Stack G, Berry M, Jin J-R, Chambon P. Functional domains of the human estrogen receptor. Cell 1987; 51: 941
1. Glick JH, Abeloff MD, Brown BW jr, Cobau CD, Johnson BL, Lichter AS, O'Bannon HB, Ottman R, Russo J, Singer K, Trafford A, Wood WC. Adjuvant chemotherapy for breast cancer. J Am Med Assn 1986; 264(24): 3461

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Pertschuk LP, Kim Y-D, Axiotis CA, Braverman AS, Carter AC, Eisenberg KB, Braithwaite 3. LV. Estrogen receptor immunocytochemistry: The promise and the perils. J Cell Biochem 1994: Suppl 19: 134.
Al Saati T, Clamons S, Cohen-Knafo E, Faye JC, Prats H, Coindre JM, Wafflart J, 4. Caveriviere P, Bayard F, Dasol G. Production of monoclonal antibodies to human estrogen receptor protein (EA) using recombinant ER (RER). Int J Cancer 1993; 56: 661
1. Esteban JM, Ahn C, Battifora H, Felder B. Quantitative immunohistochemical assay for hormonal receptors: Technical aspects and biological significance. J Cell Biochem, 1994; Suppl 19: 138
Esteban JM, Felder B, Ahn C, Simpson JF, Battifora H, Shively JE. Prognostic relevance of 6. carcinoembryonic antigen and estrogen receptor status in breast cancer patients. Cancer 1994; 74(6): 1576
Esteban JM, Ahn C, Mehta P, Battifora H. Biologic significance of quantitative estrogen 7. receptor immunohistochemical assay by image analysis in breast cancer. Am J Clin Pathol 1994; 102(2): 158
1. Esteban JM, Ahn C, Battifora H, Felder B. Predictive value of estrogen receptors evaluated by quantitative immunohistochemical analysis in breast cancer. Am J Clin Pathol 1994; 102(4) Syppl 1: S9
1. de Mascarel I, Soubevran I, MacGrogan G, Wafflart J, Bonichon F, Durand M, Avril A, Mauriac L. Troiani M. Coindre JM. Immunohistochemical analysis of estrogen receptors in 938 breast carcinomas: Concordance with biochemical assay and prognostic significance. Appl Immunohistochem 1995; 3(4): 222
Johnston SRD, Sacanni-Jotti G. Smith IE, Salter J. Newby J. Coppen M, Ebbs SR, Dowsett 10. M. Changes in estrogen receptor, progesterone receptor, and pS2 expression in tamoxifenresistant human breast cancer. Cancer Res 1996; 56; 3331
1. Veronese SM, Barbareschi M, Morelli L, Aldovini D, Mauri FA, Caffo O, Gambacorta M, Dalla Palma P. Predictive value of ER1D5 antibody immunostaining in breast cancer. Appl Immunohistochem 1996; 3(2): 86
1. Pellicer EM, Sundblad A. Evaluation antibodies to estrogen receptors. Appl Immunahistachem 1994 ; 2(2); 141
1. Pertschuk LP, Feldman JG, Kin Y-D, Braithwaite L, Schneider F, Braverman AS Axiotis C. Estrogen receptor immunocytochemistry in paraffin embedded tissues with ER1D6 predicts breast cancer endocrine response more accurately than H222Spy in frozen sections or cytosol-based ligand-binding assays. Cancer 1996; 77(12): 2514
1. Goulding H, Pinder S, Cannon P, Pearson D, Nicholson R, Snead D, Bell J, Elston CWE, Robertson JF, Blamey RW, Ellis IO. A new immunohistochemical antibody for the assessment of estrogen receptor status on routine formalin-fixed tissue samples. Hum Pathol 1995; 26: 291
1. Mauri FA, Veronese S, Frigo B, Girlando S, Losi L, Gambacorta M, Palma PD, Barbareschi M. ER1D5 and H222 (ER-ICA) antibodias to human estrogen receptor protein in breast carcinomas. Appl Immunohistochem 1994; 2(3):167
Barnes DM, Harris WH, Smith P, Millis RR, Rubens RD. Immunohistochemical determination 16. of oestrogen receptor: Comparison of different methods of assessment of staining and correlation with clinical outcome of breast cancer patients. Br J Cancer 1996; 74: 1446
1. Hopkins CA, McNeill WF, Walker GH. Comparison of antibodies for estrogen and progesterone receptors using paraffin-embedded tissues and frozen sections. Am J Clin Pathol 1996; 103(4) 603
1. Nedergaard L. Haerslev T, Jacobsen GK, Immunohistochemical study of estrogen receptors in primary breast carcinomas and their lymph node metastases including comparison of two monoclonal antibodies. APMIS 1996; 103; 20
1. Hendricks JB, Wilkerson EJ. Comparison of two antibodies for evaluation of estrogen receptors in paraffin-embedded turnors. Modern Pathol 1993; 6(6): 766

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Leong AS-Y, Milios J. Comparison of antibodies to estrogen and progesterone receptors 20. and the influence of microwave-antigen retrieval. Appl Immunohistochem 1993 ; 144: 282
Kell DL, Kamel OW, Rouse RV. Immunohistochamical analysis of broast carcinoma estrogen 21. and progesterone receptors in paraffin-ambedded tissue: Correlation of clones ER1D5 and 1 A6 with cytosol-based hormone receptor assay. Appl Immunohistochem 1993; 114): 276
Bier B, Bankfalvi A, Grote L, Blasius S, Ofner D, Bōcker W, Jasani B, Schmid KW. Wet 22. autoclave pretreatment for immunohistochemical demonstration of oestrogen receptors in routinely processed breast carcinoma tissue. Histochem J 1996; 27: 148
Sannino P, Shousha S. Demonstration of oestrogen receptors in paraffin wax sections of 23. breast carcinoma using the monoclonal antibody 105 and microwave oven processing. J Clin Pathol 1994; 47: 90 1994
1. Sacanni Jotti G, Johnston SRD, Salter J, Detre S, Dowsett M. Comparison of new immunohistochemical assay for cestrogen receptor in paraffin wax embedded breast carcinoma tissue with quantitative enzyme immunoassay. J Clin Path 1994; 47: 900
1. Balaton AJ, Mathieu M-C, Le Doussal V. Optimizaiton of heat-induced epitope retrieval for estrogen receptor determination by immunohistochemistry on paraffin sections. Appl Immunohistochem 1996; 4(4): 259
1. Barnes DM, Millis RR. Oestrogen receptors: the history, the relevance and the mathods of evaluation. In Kirkham N, Lamoine NR, eds. Progress in Pathology 2, Edinburgh, Churchill Livingstone, 1995, 89

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TABLE 1

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A-014-088	1
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Author/Journal	Studycomparison	Numberofsamples	Scoring system	2x2		Relationship to outcome
				+	H222
Mauri, et al. ApplImmunohistochem 2(3) 1994;15715	1D5 (DAKO)to H222(frozen)	374	All cells with stainednuclei were countedas positive. 7.2%(1D5+/H222-) and3.5% 1D5-/H222+.	231 + 2311D5 - 13Total 244	27103130	Specificity = 103/130 = 79.2Sensitivity = 231/244 = 94.6
Goulding, et al.Human Pathol1995; 26:29114	1D5 (DAKO)paraffin toH222, frozen	90	H scores 1D5correlation withH222 frozen R=0.7, withp<0.0001. Samecorrelation withH222 paraffin			H score of 50 = cut-off. ER 1D5correlated with outcome to TAM1D5 is more sensitive (90%vs67%) but less specific (51% vs62%) to H222
Nedergaard et al.APMIS 1995;103:2018	1D5 (DAKO)to ER-ICAfrozen and1D5 to ER-ICAparaffin	83		1D5+53- 2Total 55	72128	Weak staining, more than 10%cells corresponds to <100fmol/mgSpecificity = 0.75,sensitivity = 0.96
Pertschuk et al.Cancer 1996; 77:251413	1D5 (DAKO)to ER-ICAfrozen	74	Cut-off betweenpositive andnegative was atleast 10% of tumorcells stainingpositive	1D5+29- 14Total 43	72431	All had Stage IV disease.DemographicsWhite 38AfroAm 31Hispanic 4Asian 154 received tamoxifen ± adjuvanttherapyof 21 discordant cases, ER-1D5correctly predicted endocrineresponse in 16 cases (p < 0.02)
Author/Journal	Study comparison	Number of samples	Scoring system	2x2	Relationship to outcome
Leong and Milos.Appl.Immunohistochem 1993;1: 28220	ER 1D5(DAKO) toH222 frozen	31	No negative ERspecimens wereevaluated. 1D5correlated well withH222 on levels ofpositivity		1D5 specificity = 25/48 (73%)sensitivity = 23/26 (89%)+ Predictive value = 23/36 (64%)- Predictive value = 35/38(92%)In all cases better than H222 andER by DCC
Pellicer andSundblad, Appl.Immunohistochem 1994:2: 14112	1D5 (AMAC)to H222	300patientswith 5yearfollow-up	Staining graded 0 –3+ according to thenuclear stainingintensity	+ 1371D5 - 12149(49.7%)	1D5 was significant for DFS (p =0.01) as well as OS (p=0.01).When staining intensity wasstratified,OS reached (p = 0.007) and DFSreached (p = 0.003) for 3+intensity
Hopkins et al.Am J Clin Path1995;103: 50317	ER 1D5 toH222 paraffinand H222frozen	51		53 19098 110151 300(50.3%)	Specificity = 98/151 = 64.90%Sensitivity = 137/149 = 91.95%Discrepant results occurred withweak or heterogeneous staining.1D5 had 100% sensitivity and 88%specificity with H222 frozen
Author/Journal	Study comparison	Number of samples	Scoring system	2x2		Relationship to outcome
				DCC
				+	-
Hendricks andWilkinson ModernPath 1993; 6:76519	H222 withproteolyticdigestion ofFFPE, 1D5(AMAC) withHIER: Bothwerecompared toDCC	20		ER ICA + 86ER 1D5 + 133	1513	ER 1D5 sensitivity = 93%Specificity = 50%+ predictive value =- predictive value = 7
						ER-ICA sensitivity = 57%Specificity = 83%+ predictive value =- predictive value =

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Statement of Substantial Equivalence:

DAKO" Monoclonal Mouse Anti-Human Estrogen Receptor, 1DB (anti-ER, 1D5) is a monoclonal antibody produced as a tissue culture supernatant which is available as a concentrate (Code No. M7047) and a prediluted primary (Code No. N1575).

Anti-ER, 1D5 immunahistochemically demonstrates the presence of estrogen receptor in a variety of normal and pathological tissues. In normal tissue, anti-ER, 105 labels estrogen containing cells. Pathological reactivity includes detection of estrogen receptor in breast cancer.

Anti-ER, 1D6 is comparable in use and technology to Abbott ER-ICA Monoclonal, clone H222 which labels cells in tissues and is currently in commercial distribution. Similarities of the anti-ER, 1D5 to the Abbott ER-ICA include that both products are monoclonal antibodies directed against the human Estrogen Receptor (ER). The differences between the two products include that the DAKO anti-ER, 1D5 is directed to the N-terminal end of the ER, while the H222 clone from the Abbott ER-ICA is directed against the functional section of the ER, coded for in exon 5 of the ER DNA. Clone H222 is of rat origin, while anti-ER, 1D5 is of mouse origin. Functionally, anti-ER, 1D5 reacts with an exitope exposed in formalin-fixed, paraffin-embedded tissues by heat induced epitope retrieval. The H222 clone binds to ER found in frozen tissyes. When the H222 clone has been tested on formalin-fixed, paraffin-embedded tissues, the specificity was decreased from that seen with frozen tissyes.

Visualization of the primary antibody for the Abbott ER-ICA is achieved using an indirect peroxidase anti-peroxidase (PAP) system. ER-ICA monoclonal is located by a goat anti-rar bridging antibody. Anti-Rat PAP complex is added, which recognizes the bridging antibody. Then a precipitating enzyme generated product (hydrogen peroxide) and a chromogen substrate solution, containing DAB are added. The reaction product produces a reddish brown precipitate at the locations of the primary antibody.

The DAKO anti-estrogen receptor, clone 1D5 is substantially equivalent to the Abbott ER-ICA Monoclonal, clone H222 in that both products specifically bind to estrogen receptor proteins located in the nuclei of cells, both products require similar detection chemistry principles for visualization of the product, and both aid in the prognosis of breast carcinoma. Direct comparison Is presented in Table 2.

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TABLE 2.

DEVICE	DAKO° Monoclonal MouseAnti-Human EstrogenReceptor, clone 1D5(Product Code Nos. M7047and N1575)	Abbott ER-ICA Monoclonal,clone H222
ANTIBODY TYPE	Monoclonal, mouse origin	Monoclonal, rat origin
ISOTYPE	IgG₁, kappa
CLONE	1D5	H222
PRESENTATION	tissue culture supernatant
REACTIVITY	estrogen receptor	estrogen receptor
POSITIVE CELLTYPE	cells containing estrogen	cells containing estrogen
STAINING PATTERN	nuclear	nuclear
INTENDED USE	semi-quantitative detectionof estrogen receptor	semi-quantitative detectionof estrogen receptor
CLINICAL UTILITY	aid in prognosis andmanagement of breastcancer	aid in prognosis andmanagement of breastcancer
TECHNOLOGY	qualitative and semi-quantitativeimmunohistochemistry	qualitative and semi-quantitativeimmunohistochemistry
INTERPRETATION	light microscopy	light microscopy
SPECIMEN TYPES	paraffin embedded tissue andcryostat	cryostat
STORAGE	2.8°C	2.8°C

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Image /page/14/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle with three curved lines representing its body and wings. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the eagle. The text is in all capital letters and is black.

MAR - 3 2000

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Gretchen M. Murray, Ph.D. Regulatory Affairs Manager DAKO Corporation 6392 Via Real Carpinteria, California 93013

Re: K993957

Trade Name: DAKO® Monoclonal Mouse Anti-Human Estrogen Receptor, Clone 1D5, Antibody for Immunoenzymatic Staining (Product Code No. M7047) and DAKO® Monoclonal Mouse Anti-Human Estrogen Receptor, Clone 1D5, Ready-to-Use Antibody and Negative Control Reagent for Immunoenzymatic Staining (Product Code No. N1575)

Regulatory Class: II Product Code: MYA Dated: February 4, 2000 Received: February 17, 2000

Dear Dr. Murray:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to 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). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic OS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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Page 2

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

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Putman

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

Enclosure

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510(k) Number (if known): K993957

Device Name: Monoclonal Mouse Anti-Human Estrogen Receptor, Clone 1D5 Antibody for Immunoenzymatic Staining (DAKO Code No. M7047)

Monoclonal Mouse Anti-Human Estrogen Receptor, Clone 1D5 Ready-to-Use Antibody and Negative Control for Immunoenzymatic Staining (DAKO Code No. N1575)

Indications For Use:

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

inn of Clinical Laboratory Devi

Prescription Use V (Per 21 CFR 801.109)

IVD Use V (Per 21 CFR 801.119 Over-The-Counter Use (Per 21 CFR 801.110

(Optional Format 1-2-96)

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Regulation Number and Section

§ 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.