MOUSE ANTI-HUMAN CD3, T-CELL/FITC & CD19, B-CELL/RPE

K960531 · Dako Corp. · GKZ · Jun 14, 1996 · Hematology

Device Facts

• Record ID: K960531
• Device Name: MOUSE ANTI-HUMAN CD3, T-CELL/FITC & 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, CD3/FITC, UCHT1 + Mouse Anti-Human B-cell, CD19/RPE, HD37 (DAKO Anti-CD3/FITC and Anti-CD19/RPE) has been developed for use in flow cytometry for the analysis of T-cells and B-cells. This reagent allows simultaneous detection and quantification of total T-cells and B-cells in peripheral blood of 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 kit containing purified mouse anti-human CD3 (Clone UCHT1) conjugated with FITC and mouse anti-human CD19 (Clone HD37) conjugated with R-phycoerythrin. Used in clinical laboratories by technicians/pathologists for flow cytometric analysis of peripheral blood. Samples undergo RBC lysis; lymphocytes are gated based on morphology. Reagents bind to specific cell surface antigens (CD3 for T-cells, CD19 for B-cells). Flow cytometer detects fluorescence signals to quantify T-cell and B-cell populations. Output assists clinicians in diagnosing/monitoring immunodeficiency disorders and performing routine lymphocyte immunophenotyping.

Clinical Evidence

Clinical evaluation compared DAKO reagents to predicate Becton Dickinson Simultest CD3/CD19 using peripheral blood samples from healthy adults and ill patients. Correlation for CD3+ cells was >0.98; correlation for CD19+ cells was >0.99. Linearity testing performed using JM cells (CD3) and Raji cells (CD19) yielded r=0.999 for both. Reproducibility assessed across two flow cytometers at three antigen concentrations. Cross-reactivity with various blood cell types was evaluated.

Technological Characteristics

Reagent composition: Mouse anti-human CD3 (UCHT1) conjugated with FITC and mouse anti-human CD19 (HD37) conjugated with R-phycoerythrin in 0.05M Tris-HCl buffer (pH 7.2), 15 mM NaN3, 0.1M NaCl, 1% carrier protein. Principle: Fluorochrome-conjugated antibody binding for flow cytometric detection. Standalone reagent kit.

Indications for Use

Indicated for the analysis of T-cells and B-cells in peripheral blood of adults (healthy and ill) for immunophenotyping, including assessment of immunodeficiency disorders.

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

• Becton Dickinson Simultest CD3/CD19

Submission Summary (Full Text)

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K960531 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 8, 1995 |
| Device Name: | Mouse Anti-Human T-cell, CD3/FITC, UCHT1 +
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): | Becton Dickinson Simultest CD3/CD19 |
| Device Description: | Purified mouse anti-human CD3, Clone UCHT1, 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 NaN_{3}, 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, CD3/FITC, UCHT1 + Mouse Anti-Human B-cell, CD19/RPE, HD37 (DAKO Anti-CD3/FITC and Anti-CD19/RPE) has been developed for use in flow cytometry for the analysis of T-cells and B-cells. This reagent allows simultaneous detection and quantification of total T-cells and B-cells in peripheral blood of 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 CD3^{+} T-cells and CD19^{+} B-cells that have been previously cleared by FDA (Becton Dickinson’s Simultest CD3/CD19). When flow cytometric tests of peripheral blood samples obtained from apparently healthy adults were completed, correlation of Simultest CD3/CD19 with DAKO Anti-CD3/FITC and Anti-CD19/RPE approached a direct 1 : 1 comparison for measurement of CD3+ cells. Correlation of Simultest CD3/CD19 with DAKO Anti-CD3/FITC and Anti-CD19/RPE approached a direct 1 : 1 comparison for measurement of CD19+ |

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cells. Data for the measurement of CD3 + T-cells by DAKO Anti-CD3/FITC and Anti-CD19/RPE reagent compared to Simultest CD3/CD19 on peripheral blood samples obtained from apparently healthy adults as well as ill patients gave a correlation greater than 0.98 using the whole blood method for flow cytometry. Data for the measurement of CD19 + T-cells by DAKO Anti-CD3/FITC and Anti-CD19/RPE reagent compared to Simultest CD3/CD19 gave a correlation greater than 0.99 using the whole blood method for flow cytometry.

The CD3 antibody clone, UCHT1, was clustered at the First Leukocyte Typing Workshop, Paris, France, 1982. The CD19 antibody clone, HD37, was clustered at the Second Leukocyte Typing Workshop, Boston, 1984.

Linearity testing of DAKO CD3/FITC using JM cells gave the following linear equation:

$$
y = 0.02 + 0.98x; \ r = 0.999
$$

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

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

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-CD3/FITC and 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 First and Second Leukocyte Typing Workshops indicate that the DAKO Anti-CD3/FITC and Anti-CD19/RPE reagent performs as well as Simultest CD3/CD19 in the detection and enumeration of CD3⁺ lymphocytes and the DAKO Anti-CD3/FITC and Anti-CD19/RPE reagent performs as well as Simultest CD3/CD19 in the detection and enumeration of CD19⁺ lymphocytes using flow cytometry. Safety of the DAKO Anti-CD3/FITC plus Anti-CD19/RPE reagent and its predicate device is high as are all reagents used for in vitro testing.