MOUSE ANTI-HUMAN CD2, T-CELL/FITC AND MOUSE ANTI-HUMAN CD19, B-CELL/RPE

K961435 · Dako Corp. · GKZ · Jun 14, 1996 · Hematology

Device Facts

• Record ID: K961435
• Device Name: MOUSE ANTI-HUMAN CD2, T-CELL/FITC AND MOUSE ANTI-HUMAN CD19, B-CELL/RPE
• Applicant: Dako Corp.
• Product Code: GKZ · Hematology
• Decision Date: Jun 14, 1996
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.5220
• Device Class: Class 2

Intended Use

For In Vitro Diagnostic Use Mouse Anti-Human T-cell, CD2/FITC, MT910 + Mouse Anti-Human B-cell, CD19/RPE, HD37 (DAKO Anti-CD2/FITC and Anti-CD19/RPE) has been developed for use in flow cytometry for the analysis of CD2+ T-cells and CD19+ B-cells. This reagent allows simultaneous detection and quantification of T- and 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 using flow cytometry.

Device Story

Reagent cocktail containing purified mouse anti-human CD2 (clone MT910) conjugated to FITC and mouse anti-human CD19 (clone HD37) conjugated to RPE. Used in clinical laboratories for flow cytometric immunophenotyping of peripheral blood lymphocytes. Input: peripheral blood samples (RBCs lysed). Principle: fluorochrome-conjugated antibodies bind specific cell surface antigens (CD2, CD19); flow cytometer detects fluorescence to identify and quantify T-cell and B-cell subpopulations. Output: quantitative data on T- and B-cell populations. Assists clinicians in diagnosing and monitoring immunodeficiency disorders.

Clinical Evidence

Clinical evaluation using peripheral blood samples from healthy adults and patients. Compared combination reagent to individual predicate reagents (K945692, K943284). Results: correlation >0.99 for both CD2+ and CD19+ cell counts. Linearity testing (JM cells for CD2, Raji cells for CD19) showed r=0.9965 and r=0.999 respectively. Reproducibility assessed across two flow cytometers at three antigen concentrations. Cross-reactivity with various blood cell types evaluated.

Technological Characteristics

Reagent cocktail: purified mouse anti-human CD2 (MT910) conjugated to FITC and CD19 (HD37) conjugated to RPE. Buffer: 0.05M Tris-HCl, pH 7.2, 15 mM NaN3, 0.1M NaCl, 1% carrier protein. Analyte detection via flow cytometry fluorescence. No software or electronic components; reagent-based diagnostic.

Indications for Use

Indicated for the analysis of CD2+ T-cells and CD19+ B-cells in peripheral blood specimens from adults, including those with immunodeficiency disorders, using flow cytometry.

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 Monoclonal Mouse Anti-Human T-cell, CD2/FITC, MT910 (K945692)
• DAKO Monoclonal Mouse Anti-Human B-cell, CD19/RPE, Clone HD37 (K943284)

Submission Summary (Full Text)

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K961435 JUN 14 1996

510(k) Summary

Submitter: DAKO Corporation
6392 Via Real
Carpinteria, CA 93013
[phone]

Contact: Gretchen M. Murray, Ph.D., Regulatory Affairs Asst. Manager

Date Summary
Prepared: December 27, 1995

Device Name: Mouse Anti-Human T-cell, CD2/FITC, MT910 +
Mouse Anti-Human B-cell, CD19/RPE, HD37

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

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

Product Code: GKZ

Predicate Device(s): DAKO Monoclonal Mouse Anti-Human T-cell, CD2/FITC, MT910 (DAKO Code Number F0767, FDA K945692) and
DAKO Monoclonal Mouse Anti-Human B-cell, CD19/RPE,
Clone HD37 (DAKO Code Number R0808, FDA K943284)

Device Description: Purified mouse anti-human CD2, Clone MT910, conjugated with fluorescein isothiocyanate, isomer 1 (FITC) + purified mouse anti-human CD19, Clone HD37, conjugated with R-phycoerythrin, present in 0.05M Tris-HCl buffer, pH 7.2, 15 mM NaN₃, 0.1M NaCl, stabilized with 1% carrier protein.

Subpopulations of lymphocytes may be stained with fluorochrome-conjugated antibody and evaluated in peripheral blood specimens when contaminating red blood cells (RBC's) are lysed prior to flow cytometric analysis. A subpopulation of WBC's are selected for assessment based upon cell morphology.

Intended Use: For In Vitro Diagnostic Use

Mouse Anti-Human T-cell, CD2/FITC, MT910 + Mouse Anti-Human B-cell, CD19/RPE, HD37 (DAKO Anti-CD2/FITC and Anti-CD19/RPE) has been developed for use in flow cytometry for the analysis of CD2⁺ T-cells and CD19⁺ B-cells. This reagent allows simultaneous detection and quantification of T- and 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 using flow cytometry.

Comparison of
Technological
Characteristics

Performance characteristics have been established by clinical evaluation of compared to the individual single reagent predicate devices that quantitatively measure CD2⁺ T-cells and CD19⁺ B-cells that have been previously cleared by FDA (DAKO CD2/FITC, Code No. F0767 and DAKO CD19/RPE, Code No. R0808). When flow cytometric tests of peripheral blood samples obtained from apparently healthy adults were completed, correlation of Anti-CD2, MT910 with

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DAKO Anti-CD2/FITC and Anti-CD19/RPE approached a direct 1 : 1 comparison for measurement of CD2+ cells. Correlation of Anti-CD19, HD37 with DAKO Anti-CD2/FITC and Anti-CD19/RPE approached a direct 1 : 1 comparison for measurement of CD19+ cells. Data for the measurement of CD2+ T-cells by DAKO Anti-CD2/FITC and Anti-CD19/RPE reagent compared to DAKO CD2/FITC gave a correlation greater than 0.99 using the whole blood method for flow cytometry on peripheral blood samples from normal and ill patients. Data for the measurement of CD19+ T-cells by DAKO Anti-CD2/FITC and Anti-CD19/RPE reagent compared to DAKO CD19/RPE gave a correlation greater than 0.99 using the whole blood method for flow cytometry on peripheral blood samples from normal and ill patients.

The CD2 antibody clone, MT910, was clustered at the Second Leukocyte Typing Workshop, Boston, 1984. The CD19 antibody clone, HD37, was clustered at the Second Leukocyte Typing Workshop, Boston, 1984.

Linearity testing of DAKO CD2/FITC using JM cells gave the following linear equation:

$$
y = -0.02 + 0.997x; \quad r = 0.9965
$$

Linearity testing of DAKO CD19/RPE using Raji cells gave the following linear equation:

$$
y = -0.49 + 0.99x; \quad r = 0.999
$$

In addition, reproducibility of DAKO reagents using replicates (from peripheral blood) run on two different flow cytometers was measured at three concentrations of each antigen. Cross-reactivity of Anti-CD2/FITC, plus Anti-CD19/RPE with peripheral blood cells (red blood cells, monocytes, granulocytes, lymphocytes, and platelets) was measured.

## Conclusions:

Results of the above testing as well as the information provided by the Second Leukocyte Typing Workshop indicate that the DAKO Anti-CD2/FITC plus Anti-CD19/RPE reagent performs as well as DAKO CD2/FITC in the detection and enumeration of CD2+ lymphocytes while the DAKO Anti-CD2/FITC plus Anti-CD19/RPE reagent performs as well as DAKO CD19/RPE in the detection and enumeration of CD19+ lymphocytes using flow cytometry. Safety of the DAKO Anti-CD2/FITC plus Anti-CD19/RPE reagent and its individual predicate devices is high as all reagents are used for in vitro testing.

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