For In Vitro Diagnostic Use. The Prostate-63 Cancer Diagnostic Test features a mouse monoclonal antibody, clone 4A4, that recognizes the human p63 protein in the nucleus of prostatic basal cells and urothelial tissues. This test is intended for laboratory use to qualitatively identify by immunohistochemistry the p63 antigen in histological sections from formalin-fixed paraffin-embedded tissue of normal and/or pathological prostate tissue obtained by needle biopsy or surgical procedures. The presence or absence of p63 staining aids the pathologist in the differential diagnosis of prostate cancer in conjunction with morphological findings seen with hematoxylin and eosin staining complemented by proper controls. The clinical interpretation of any staining or its absence should be evaluated within the context of the patient's clinical history and other diagnostic tests by a qualified pathologist.

Device Description

The Prostate-63 Cancer Diagnostic Test features a mouse monoclonal antibody, clone 4A4, that recognizes the human p63 protein in the nucleus of prostatic basal cells and urothelial tissues. The 4A4 anti-p63 monoclonal antibody is provided at a working dilution in a buffer containing 50 mM Tris-HCl, pH 7.2, and 15 mM sodium azide. The immunogen is p63 recombinant protein comprising amino acids 1-205 of the N-terminal portion of human △Np63 protein.

AI/ML Overview

The provided text describes the intended use and background of the Prostate-63 Cancer Diagnostic Test, as well as a list of references to relevant studies. However, it does not contain a dedicated section detailing specific acceptance criteria or an explicit study proving the device meets those criteria in a structured manner typical of a regulatory submission.

Instead, the performance characteristics are discussed by referencing several scientific publications. The acceptance of this device by the FDA (as indicated by the 510(k) clearance letter) implies that the provided data (including these referenced studies) were deemed sufficient to demonstrate substantial equivalence to a predicate device.

Based on the provided text, here's what can be extracted and inferred:

1. Table of Acceptance Criteria and Reported Device Performance:

Since explicit acceptance criteria are not stated, the table below will summarize the implied performance based on the claims and references provided. The "acceptance criteria" here are interpreted as the expected performance of a basal cell marker for prostate cancer diagnosis.

Acceptance Criteria (Inferred from clinical utility)	Reported Device Performance (as per references cited)
Stains prostatic basal cells of normal glands in needle biopsies with high sensitivity.	Sensitivity approaching 100% (Wu et al., 2004).
Stains benign glands derived from TURP.	Positive in 11 of 12 (95%) sections (Shah et al., 2002).
Does not stain neuroendocrine or luminal, secretory cells of the prostate.	Signoretti et al., 2000.
Basal cells are absent from invasive prostate carcinoma.	Hedrick and Epstein, 1995 (cited in a longer version of the extract not provided in full in the prompt) / General understanding of prostate carcinoma pathology.
Offers advantages over traditional basal cell markers (e.g., diffuse cytoplasmic stain vs. distinct nuclear stain).	Dark nuclear staining pattern (Shah et al., 2002).
Recognizes all known p63 protein isoforms on western blots and shows strong nuclear staining in relevant cells.	Yang et al., 1998.
Recognizes basal cells in mouse tissues (skin, breast, prostate, urothelia) but not in p63-deficient embryos.	Yang et al., 1999.

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

The document primarily references external studies rather than detailing a single "test set" for this specific 510(k) submission. Therefore, the sample sizes and provenance would be varied across the referenced papers.

Wu et al., 2004: "100 consecutive prostate carcinoma diagnosed by needle biopsies." (Implies retrospective data, likely from a single institution, but country of origin not specified).
Shah et al., 2002: "11 of 12 (95%) sections derived from transurethral resection of the prostate (TURP)." (Small sample, likely retrospective, country of origin not specified).
Other cited studies (Signoretti et al., 2000; Weinstein et al., 2002; Shah et al., 2004; Davis et al., 2002; Yang et al. publications) would have their own sample sizes and methodologies, which are not detailed in this summary.

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

This information is not provided in the 510(k) summary. Given that the references are peer-reviewed publications, it's highly probable that pathologists were involved in establishing diagnoses and ground truth, but their number and specific qualifications are not stated in this document.

4. Adjudication Method:

This information is not provided in the 510(k) summary.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study:

No MRMC study is explicitly mentioned as part of the 510(k) submission. The document discusses the utility of the antibody in facilitating diagnosis and offers advantages over other markers, implying improved effectiveness for pathologists, but a formal MRMC study calculating an effect size of human readers with vs. without AI assistance (which is not applicable here as it's an antibody, not an AI) is not presented.

6. Standalone (Algorithm Only) Performance:

This is not an AI/algorithm-based device, but rather an immunohistochemical reagent. Therefore, the concept of "standalone performance" for an algorithm is not applicable. The performance is inherently tied to human interpretation by a pathologist.

7. Type of Ground Truth Used:

The ground truth for the referenced studies and the intended use of this diagnostic test is based on pathology findings (morphological findings seen with hematoxylin and eosin staining, and the specific characteristics of prostate cancer identified by qualified pathologists).

8. Sample Size for the Training Set:

The document describes the origin and specificity of the antibody (Immunogen: p63 recombinant protein, tested on BHK cells, mouse tissues), but it does not describe a "training set" in the context of machine learning or an algorithm. The development of the antibody involved biological and laboratory research, which would implicitly include various experiments and testing but not a formal "training set" as understood for AI.

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

Again, for an antibody reagent, there isn't a "training set" with established ground truth in the AI sense. The antibody's specificity and reactivity were established through:

Immunogen: p63 recombinant protein.
Testing on BHK cells expressing p63 cDNAs vs. control vectors.
Immunohistochemistry in mouse tissues, comparing staining in p63-positive tissues with those from p63-deficient embryos (Yang et al., 1999).
Western blot analysis for recognition of p63 protein isoforms (Yang et al., 1998).

These methods established the biological specificity and performance characteristics of the 4A4 anti-p63 monoclonal antibody.

Summary

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K050063

510 (k) Summary

Asymmetrx, Inc. 117 Senate Brook Drive Amston, CT 06231 (860) 716-8888

Contact Person: Maria McKeon

Date Summary Prepared: February 4, 2005

Name of Device:

1. Proprietary/Trade Name: Prostate-63 Cancer Diagnostic Test
1. Common Name: antibody to p63 protein in basal cells
1. Classification Name: Immunohistochemical reagent, antibody (monoclonal or polyclonal) to p63 protein in nucleus of prostatic basal cells

DESCRIPTION OF DEVICE:

Intended Use

Background

p63 is a member of the p53 family, which also includes p73 (Yang et al., 1998; Yang and McKeon, 2000). p63 is strongly expressed in the basal or progenitor cells of a large number of epithelial tissues (Yang et al., 1998). These tissues include squamous, transitional, and glandular epithelia, such as prostate, breast, esophagus, bladder, airway, and epidermis, among others. In general, these p63 positive basal cells act as the progenitors or reserve cells in these regenerative epithelia (Parsa et al., 1999). p63deficient mice lack all epithelial tissues in which p63 is highly expressed, such as prostate, breast and epidermis. p63 mutant mice also show severe defects in limb and craniofacial development (Yang et al., 1999). A similar pattern is observed in patients with EEC syndrome, an autosomal dominant disorder characterized by ectodactyly, ectodermal dysplasia, and facial clefts, where affected patients are found to have dominant heterozygous p63 mutations (Celli et al., 1999). Recent studies support the utility of the 4A4 anti-p63 monoclonal antibody in the differential diagnosis of prostate

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cancer. (Signoretti et al., 2000; Weinstein et al., 2002; Shah et al., 2002; Davis et al., 2002; Shah et al., 2004).

Reagent provided.

The 4A4 anti-p63 monoclonal antibody is provided at a working dilution in a buffer containing 50 mM Tris-HCl, pH 7.2, and 15 mM sodium azide.

Immunogen.

p63 recombinant protein comprising amino acids 1-205 of the N-terminal portion of human △Np63 protein (Yang et al., 1998).

Precautions:

1. For professional users only.
1. This product contains sodium azide (NaN}), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing.
1. As with any product derived from biological sources, proper handling procedures should be used.

The specifics of Specimen Preparation, Staining Procedure and Storage Requirements are detailed in the label.

Performance Characteristics.

The Prostate-63 Cancer Diagnostic Test was developed as a tool to facilitate the diagnosis of prostate cancer. Recent studies have supported the utility of the 4A4 antip63 monoclonal antibody in prostate cancer diagnosis (Signoretti et al., 2000; Weinstein et al., 2002; Shah et al., 2002; Davis et al., 2002; Shah et al., 2004). The basis of the Prostate-63 Cancer Diagnostic Test is the use of the 4A4 anti-p63 monoclonal antibody that recognizes all known p63 protein isoforms. Basal cells of the prostate show dark staining with the p63 antibody whereas secretory and neuroendocrine cells of the prostate typically show no staining (Yang et al., 1998; Signoretti et al., 2000). The dark nuclear staining pattern of the p63 antibody in basal cells may offer advantages over traditional basal cell markers which yield a diffuse cytoplasmic stain (Shah et al., 2002),

The 4A4 anti-p63 monoclonal antibody recognizes all forms of p63 protein on western blots, and shows strong nuclear staining of baby hamster kidney (BHK) cells expressing p63 cDNAs but not BHK cells expressing control vectors (Y ang et al., 1998). By immunohistochemistry in mouse tissues, the 4A4 anti-p63 monoclonal antibody recognizes basal cells of skin, breast, prostate, urothelia, among other tissues, but does not show significant staining in corresponding tissues of embryos lacking the p63 gene (Yang et al., 1999)

p63 stains prostatic basal cells of normal glands in needle biopsies with a sensitivity approaching 100% (Wu et al., 2004). Benign glands were also positive in 11 of 12 (95%) sections derived from transurethral resection of the prostate (TURP; Shah et al., 2002). p63 does not stain neuroendocrine or luminal, secretory cells of the prostate (Signoretti et al., 2000). As basal cells are absent from invasive prostate carcinoma (Hedrick and

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cancer. (Signoretti et al., 2000; Weinstein et al., 2002; Shah et al., 2002; Davis et al., 2002; Shah et al., 2004).

Reagent provided.

The 4A4 anti-p63 monoclonal antibody is provided at a working dilution in a buffer containing 50 mM Tris-HCl, pH 7.2, and 15 mM sodium azide.

Immunogen.

p63 recombinant protein comprising amino acids 1-205 of the N-terminal portion of human △Np63 protein (Yang et al., 1998).

Precautions:

1. For professional users only.
1. This product contains sodium azide (NaN}), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing.
1. As with any product derived from biological sources, proper handling procedures should be used.

The specifics of Specimen Preparation, Staining Procedure and Storage Requirements are detailed in the label.

Performance Characteristics.

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cancer. (Signoretti et al., 2000; Weinstein et al., 2002; Shah et al., 2002; Davis et al., 2002; Shah et al., 2004).

Reagent provided.

The 4A4 anti-p63 monoclonal antibody is provided at a working dilution in a buffer containing 50 mM Tris-HCl, pH 7.2, and 15 mM sodium azide.

Immunogen.

p63 recombinant protein comprising amino acids 1-205 of the N-terminal portion of human △Np63 protein (Yang et al., 1998).

Precautions:

1. For professional users only.
1. This product contains sodium azide (NaN}), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing.
1. As with any product derived from biological sources, proper handling procedures should be used.

The specifics of Specimen Preparation, Staining Procedure and Storage Requirements are detailed in the label.

Performance Characteristics.

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Shah RB, Zhou M, LeBlanc M, Snyder M, Rubin MA. Comparison of the basal cellspecific markers, 34betaE12 and p63, in the diagnosis of prostate cancer. Am J Surg Pathol. 2002 Sep:26(9):1161-8.

Shah RB, Kunju LP, Shen R, LeBlanc M, Zhou M, Rubin MA. Usefulness of basal cell cocktail (34betaE12 + p63) in the diagnosis of atypical prostate glandular proliferations. Am J Clin Pathol. 2004 Oct;122(4):517-23.

Signoretti S. Waltregny D. Dilks J. Isaac B. Lin D. Garraway L. Yang A. Montironi R. McKeon F. Loda M. p63 is a prostate basal cell marker and is required for prostate development. Am J Pathol. 2000 Dec:157(6):1769-75.

Weinstein MH, Signoretti S, Loda M. Diagnostic utility of immunohistochemical staining for p63, a sensitive marker of prostatic basal cells. Mod Pathol. 2002 Dec; 15(12): 1302-1308.

Wu HH, Lapkus O, Corbin M. Comparison of 34betaE12 and P63 in 100 consecutive prostate carcinoma diagnosed by needle biopsies.

Appl Immunohistochem Mol Morphol. 2004 Dec:12(4):285-9.

Yang A, Kaghad M, Wang Y, Gillett E, Fleming MD, Dotsch V, Andrews NC, Caput D, McKeon F. p63, a p53 homolog at 3g27-29, encodes multiple products with transactivating, death-inducing, and dominant-negative activities. Mol Cell. 1998 Sep;2(3):305-16.

Yang A, McKeon F. P63 and P73: P53 mimics, menaces and more. Nat Rev Mol Cell Biol. 2000 Dec;1(3):199-207. Review.

Yang A, Schweitzer R, Sun D, Kaghad M, Walker N, Bronson RT, Tabin C, Sharpe A, Caput D. Crum C. McKeon F. p63 is essential for regenerative proliferation in limb. craniofacial and epithelial development. Nature. 1999 Apr 22;398(6729):714-8.

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/5/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract symbol resembling three stylized human figures or birds in flight, stacked on top of each other.

FEB - 9 2005

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Ms. Maria F. McKeon President AsymmetRx, Inc. 117 Senate Brook Drive Amston, Connecticut 06231

B - 9 2005

Re: K050063

Trade/Device Name: Prostate-63 Cancer Diagnostic Test Regulation Number: 21 CFR § 864.1860 Regulation Name: Immunohistochemistry Regulatory Class: I Product Code: NTR Dated: December 28, 2004 Received: January 12, 2005

Dear Ms. McKeon:

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 Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); and 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 information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (240) 276-0484. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). 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 (301) 443-6597 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html

Sincerely yours,

lobatz Beckerh

Robert L. Becker, Jr., MD, PM Director Division of Immunology and Hematology 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): K050063

Device Name: Prostate-63 Prostate Cancer Diagnostic Test

Indications For Use:

Dated this 4th day of February, 2005

Maria F. McKeon, President AsymmetRx, Inc.

Prescription Use (Part 21 CFR 801 Subpart D)

AND/OR

Over-The-Counter Use (21 CFR 807 Subpart C)

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

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (O[VD)

-maria Elam

ision Sion-Off) (Division of Clinical Laboratory Devices

510(k) Number_K 50063

Page 1 of 1

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.