DAKO MOUSE ANTI-HUMAN B-CELL, CD19/RPE-CY5, CLONE HD37

K980635 · Dako Corp. · GKZ · Jul 22, 1998 · Hematology

Device Facts

• Record ID: K980635
• Device Name: DAKO MOUSE ANTI-HUMAN B-CELL, CD19/RPE-CY5, CLONE HD37
• Applicant: Dako Corp.
• Product Code: GKZ · Hematology
• Decision Date: Jul 22, 1998
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.5220
• Device Class: Class 2

Intended Use

Monoclonal Mouse Anti-Human B-cell, CD19, Clone HD37, RPE-Cy5 conjugated, has been developed for use in flow cytometry for the analysis of B-cells. This reagent allows simultaneous detection and quantification of CD19-positive cells (B-cells) in normal and pathological conditions such as immunodeficiency disorders. It is one component of the suggested monoclonal antibody (MAb) combinations for routine immunophenotyping of lymphocytes in peripheral blood. Immunophenotyping of lymphocytes is widely applied for detection and classification of hematovojetic malignancies, and for diagnosis of immunodeficiencies. DAKO Anti-CD19/Cy5 is one of the reagents utilized when performing immunophenotyping of lymphocytes.

Device Story

Reagent consists of monoclonal mouse anti-human CD19 antibody (clone HD37) conjugated to R-phycoerythrin (RPE) and cyanin 5 (Cy5). Used in clinical laboratories by trained personnel for flow cytometry analysis of peripheral blood samples. Antibody binds specifically to CD19 antigens on B-lymphocytes; RPE-Cy5 fluorochrome allows detection via flow cytometer. Clinicians use quantification of CD19+ cells to assist in diagnosing immunodeficiency disorders and classifying hematologic malignancies. Proper gating during flow cytometry excludes non-target cells like monocytes. Provides standardized immunophenotyping data to support clinical decision-making regarding patient immune status and disease classification.

Clinical Evidence

Bench testing only. Reproducibility assessed across two flow cytometer platforms using high, medium, and low CD19+ concentration samples; CVs ranged from 1.37% to 4.11%. Specificity verified in five healthy donors; antibody binding specific to lymphocytes (13.34% average) with minimal cross-reactivity to other blood components. Correlation study compared subject device to predicate in 180 samples (153 healthy, 27 pathological), yielding a linear regression equation of y = 0.45 + 0.98x and R^2 = 0.8832, demonstrating 1:1 comparability.

Technological Characteristics

Monoclonal mouse anti-human CD19 antibody (clone HD37) conjugated to RPE-Cy5. Supplied in 0.05M Tris-HCl buffer, pH 7.2, 15mM NaN3, 0.1M NaCl, 1% carrier protein. Sensing principle: fluorescence-based flow cytometry. Intended for use with standard flow cytometers. No electronic or software components integral to the reagent itself.

Indications for Use

Indicated for in vitro diagnostic use in flow cytometry for the analysis of B-cells in peripheral blood. Used for the detection and quantification of CD19-positive cells in patients with normal or pathological conditions, including immunodeficiency disorders and hematologic malignancies.

Regulatory Classification

Identification

An automated differential cell counter is a device used to identify one or more of the formed elements of the blood. The device may also have the capability to flag, count, or classify immature or abnormal hematopoietic cells of the blood, bone marrow, or other body fluids. These devices may combine an electronic particle counting method, optical method, or a flow cytometric method utilizing monoclonal CD (cluster designation) markers. The device includes accessory CD markers.

Special Controls

*Classification.* Class II (special controls). The special control for this device is the FDA document entitled “Class II Special Controls Guidance Document: Premarket Notifications for Automated Differential Cell Counters for Immature or Abnormal Blood Cells; Final Guidance for Industry and FDA.”

Predicate Devices

• DAKO Mouse anti-human B-cell, CD19, RPE, clone HD37, Code No. R0808

Submission Summary (Full Text)

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.111 22 1998

K980635

510(k) Summary

Submitter:

DAKO Corporation 6392 Via Real Carpinteria, CA 93013 805-566-6655

Contact:

Gretchen M. Murray, Ph.D.

Date Summary Prepared:

January 2, 1998

Device Name:

DAKO® Mouse Anti-Human B-cell, CD19/RPE-Cy5, Clone HD37 (Product Code No. C7066

Device

Class II according to 21 CFR 864.5220, on the basis that monoclonal Classification: antibodies are accessories for automated differential cell counters.

Panel: The device classification is under the Hematology and Pathology Devices panel. Division of Clinical Laboratory Devices.

DAKO Mouse anti-human B-cell, CD19/RPE, clone HD37, Code No. R0808 Predicate Device:

Monoclonal Mouse Anti-Human B-cell, CD19, RPE-Cy5-conjugated, Clone Device HD37 is specific for B-lymphocyte cluster determinants as evaluated by the Description: International Workshop on Human Leukocyte Differentiation Antigens. HD37 CD19-specific monoclonal antibody was designated B28 antibody at the Second Workshop (Reinherz, EL, Haynes, BF, Nadler, LM, Bernstein, ID, Leukocyte Typing II, Vol. 2. New York-Berlin-Heidelberg-Tokyo: eds. Springer Verlag, 1986.) Purified monoclonal mouse anti-human CD19 is produced in tissue culture, dialyzed and conjugated with R-phycoerythrin (RPE) covalently coupled to cyanin 5 (Cy5). One ml (1,0 ml) containing the conjugated antibody is supplied in 0.05M Tris-HCI buffer, pH 7.2, 15mM NaN3, 0.1M NaCl, stabilized with 1% carrier protein.

Intended Use:

For In Vitro Diagnostic Use

Monoclonal Mouse Anti-Human B-cell, CD19, RPE-Cy5 conjugated, Clone HD37, (Anti-CD19/Cy5) has been developed for use in flow cytometry for the analysis of B-cells in peripheral blood. This reagent allows simultaneous detection and quantification of CD19-positive cells (B-cells) in normal and pathological conditions such as immunodeficiency disorders. It is one component of the suggested monoclonal antibody (MAb) combinations for routine immunophenotyping of lymphocytes in peripheral blood.

Comparison of Technological Characteristics

Binding linearity was determined over serial dilutions of a cell line known to express the antigen diluted with a cell line that has no antigenic sites. For Anti-CD19/Cy5, the cell line with known antigenic reactivity is Raji cells,

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while the cell line without antigenic sites is JM cells. Five dilutions were tested, with a linear equation calculated from the results. The equation for Anti-CD19/Cy5, HD37 was $y = 0.01% + 0.98x$. $r^2 = 0.999$.

Ten replicates from peripheral blood of three donors were tested for reproducibility of Anti-CD19/RPE-Cy5 and run on two flow cytometers from different manufacturers at three concentrations of antigen. Different levels of CD19+ lymphocytes were selected from a population of normal and abnormal peripheral blood samples. Each level of CD19 was analyzed within one day on both machines.

| FACScan | | Mean % CD19 + | ± 1 SD | %CV | n |
|------------|--------------|---------------|--------|------|----|
| | High Level | 90.87 | 1.24 | 1.37 | 10 |
| | Medium Level | 51.28 | 1.47 | 2.87 | 10 |
| | Low Level | 26.77 | 1.02 | 3.81 | 10 |
| Profile II | | Mean % CD19 + | ± 1 SD | %CV | n |
| | High Level | 87.16 | 2.09 | 2.40 | 10 |
| | Medium Level | 49.75 | 1.54 | 3.09 | 10 |
| | Low Level | 26.13 | 1.08 | 4.11 | 10 |

Specificity of Anti-CD19/Cy5 has been verified by tests performed on five apparently healthy adult donors of various races at DAKO Corporation. Cell populations tested were RBC's, granulocytes, monocytes, lymphocytes and platelets. The results indicate antibody binding of Anti-CD19/Cy5 is specific for lymphocytes. Lymphocytes bound to Anti-CD19/Cy5 antibodies on an average of 13.0%, representative of the B-cell population. Approximately 7% of monocytes bound with the Anti-CD19/Cy5. However, monocyte binding can be excluded from the lymphocyte analysis by proper gating on lymphocytes.

| | %Positive Red<br>Blood Cells | % Positive<br>Granulocytes | % Positive<br>Monocytes | % Positive<br>Lymphocytes | % Positive<br>Platelets |
|--------------------------|------------------------------|----------------------------|-------------------------|---------------------------|-------------------------|
| Average (n=5)<br>(range) | 0.04<br>(0.0-0.2) | 1.10<br>(0.3-1.7) | 7.66<br>(3.5-9.7) | 13.34<br>(10.1-17.6) | 0.22<br>(0.0-0.5) |

DAKO Anti-CD19/Cy5 Specificity

Correlation of Anti-CD19/Cy5, HD37 to a predicate Anti-CD19/RPE, HD37 reagent, was determined by testing duplicate samples with each reagent across 153 normal, apparently healthy individuals at three geographically separate laboratories. Linear regression analysis of the data gave the following equations and Pearson correlations.

$y_{\text{(DAKO CD19/Cy5+ Lymphocytes)}} = 2.27 + 0.77 x_{\text{(DAKO CD19/RPE + Lymphocytes)}}$. $R^2 = 0.6743$. $n = 153$

> In addition, 27 samples from patients with illnesses were compared, and their data added to the results of the testing of the 153 apparently healthy individuals. Linear correlation was performed on the total database. Linear regression analysis gave the following equation and R2:

$y_{\text{(DAKO CD19/Cy5+ Lymphocytes)}} = 0.45 + 0.98 x_{\text{(DAKO CD19/RPE + Lymphocytes)}}$.

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$$\begin{array}{rcl} \mathsf{R}^2 &=& \mathsf{0.8832} \\ \mathsf{n} &=& \mathsf{1800}. \end{array}$$

.

:

:

This equation indicates that Anti-CD19/Cy5, HD37 reagent and the Anti-CD19/RPE, HD37 reagent are comparable on a 1:1 basis.

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Image /page/3/Picture/2 description: The image is a black and white logo for the U.S. Department of Health and Human Services. The logo features a stylized image of an eagle with its wings spread, and the words "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" are arranged in a circle around the eagle. The eagle is composed of three thick, curved lines that suggest movement and flight. The overall design is simple and clean, conveying a sense of authority and professionalism.

JUL 22 1998

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Gretchen M. Murray, Ph.D., RAC Requlatory Affairs Manager DAKO CORPORATION 6392 Via Real Carpinteria, CA 93013

K980635 Re: Trade Name: Mouse Anti-human B-cell, CD19/RPE-Cy5, Clone HD37 Regulatory Class: II Product Code: GKZ Dated: May 28, 1998 · · · Received: June 1, 1998

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 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 requlations 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 requirement, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic (QS) 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 Reqister. 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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Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at [phone].

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 [phone]. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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 [phone] or at [phone] or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours,

Steven Sutman

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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Page l of l

510(k) Number (if known): K9801635

Device Name: _ Monoclonal Mouse Anti-Human B-cell, CD19 Clone HD37 RPE-Cy5 Conjugated

## Indications For Use:

Monoclonal Mouse Anti-Human B-cell, CD19, Clone HD37, RPE-Cy5 conjugated, has been developed for use in flow cytometry for the analysis of B-cells. This reagent allows simultaneous detection and quantification of CD19-positive cells (B-cells) in normal and pathological conditions such as immunodeficiency disorders. It is one component of the suggested monoclonal antibody (MAb) combinations for routine immunophenotyping of lymphocytes in peripheral blood.

Immunophenotyping of lymphocytes is widely applied for detection and classification of hematovojetic malignancies, and for diagnosis of immunodeficiencies. DAKO Anti-CD19/Cy5 is one of the reagents utilized when performing immunophenotyping of lymphocytes.

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

OR

Over-The-Counter Use (Per 21 CRF 801.110)

Prescription Use V (Per 21 CFR 801.109)

IVD Use (Per 21 CFR 801.119)

(Optional Format 1-2-96)