CD3 FITC/CD19 R-PE/CD45 TRI-COLOR MONOCLONAL ANTIBODY COMBINATION
Device Facts
| Record ID | K964856 |
|---|---|
| Device Name | CD3 FITC/CD19 R-PE/CD45 TRI-COLOR MONOCLONAL ANTIBODY COMBINATION |
| Applicant | Caltag Laboratories, Inc. |
| Product Code | GKZ · Hematology |
| Decision Date | Jan 17, 1997 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 864.5220 |
| Device Class | Class 2 |
Indications for Use
CALTAG CD3/CD19/CD45 is a fluorescent reagent containing a combination of CD3, CD19 and CD45 monoclonal antibodies conjugated to fluorescein, phycoerythrin and the tandem fluorochrome PE-Cy5, respectively. This reagent permits the simultaneous identification of CD3+ mature T lymphocytes, CD19+ mature B lymphocytes, and CD45+ leukocytes including lymphocytes, monocytes and granulocytes, by flow cytometric methods.
Device Story
Device is a fluorescent monoclonal antibody cocktail (CD3-FITC, CD19-R-PE, CD45-TRI-COLOR) for flow cytometry. Input: peripheral blood leukocytes. Process: incubation with antibodies; lysis of red blood cells; washing; fixation. Principle: antibodies bind specific cell surface antigens (CD3, CD19, CD45). Output: fluorescently labeled cells analyzed by flow cytometer. Used in clinical laboratories by technicians/pathologists. Output allows identification/quantification of lymphocyte subsets; aids in immunophenotyping and clinical assessment of immune status.
Clinical Evidence
Bench testing only. Study of 155 healthy donors (ages 16-72) established expected values for CD3, CD19, and CD45 components. Correlation studies (n=175) compared the combination reagent against single-antibody reagents, showing high correlation (r² 0.948-0.989) and slopes near 1.0. Specificity data provided across ethnic groups (Caucasian, Black, Hispanic, Oriental) confirming consistent staining of target leukocyte populations.
Technological Characteristics
Mouse monoclonal antibody combination conjugated to FITC, R-PE, and PE-Cy5 (TRI-COLOR). Liquid form in PBS. Analyzed via flow cytometry. No specific materials or software algorithms described; relies on standard immunofluorescence and flow cytometric gating principles.
Indications for Use
Indicated for the simultaneous identification of CD3+ mature T lymphocytes, CD19+ mature B lymphocytes, and CD45+ leukocytes (lymphocytes, monocytes, granulocytes) in human peripheral blood samples via 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
- Caltag CD3 FITC single monoclonal antibody
- Caltag CD19 R-PE single monoclonal antibody
- Coulter CD19 RD1 single monoclonal antibody
Submission Summary (Full Text)
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CALTAG LABORATORIES
JAN 17 1997
K964856
510(K) SUMMARY
SUMMARY OF SAFETY AND EFFECTIVENESS DATA
CD3 FITC/CD19 R-PE/CD45 TRI-COLOR™
Mouse Monoclonal Antibody Combination
To Human Cell Surface Antigens by Flow Cytometry
NAME AND LOCATION OF MANUFACTURER:
Caltag Laboratories, Inc.
1849 Old Bayshore Highway
Suite 200
Burlingame, CA 94010
(800) 874-4007
NAME OF CONTACT PERSON:
Robert C. Johnson
Executive Vice President
Caltag Laboratories, Inc.
DATE OF PREPARATION OF SUMMARY:
December 1, 1996
Caltag Laboratories Inc. • 1849 Bayshore Blvd., Suite #200 • Burlingame, CA 94010
Telephone (415) 652-0468 • (800) 874-4007 • FAX (415) 652-9030
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## TRADE NAME OF THE DEVICE:
CD3 FITC/CD19 R-PE/CD45 TRI-COLOR™
Mouse Monoclonal Antibody Combination
To Human Cell Surface Antigens by Flow Cytometry
## COMMON NAME:
Caltag CD3 FITC/CD19 R-PE/CD45 TRI-COLOR™ Monoclonal Antibody Combination
## CLASSIFICATION NAME:
Automated Differential Cell Coulter (21 CFR 864.5220)
## LEGALLY MARKETED DEVICE (PREDICATE DEVICE) TO WHICH THE MANUFACTURER IS CLAIMING SUBSTANTIAL EQUIVALENCE:
The Caltag CD3 FITC monoclonal antibody in the CD3 FITC/CD19 R-PE/CD45 TRI-COLOR combination is substantially equivalent to the Caltag CD3 FITC single monoclonal antibody for in vitro diagnostic use.
The Caltag CD19 R-PE monoclonal antibody in the CD3 FITC/CD19 R-PE/CD45 TRI-COLOR combination is substantially equivalent to the Caltag CD19 R-PE single monoclonal antibody for in vitro diagnostic use, and is substantially equivalent to the Coulter CD19 RD1 single monoclonal antibody for in vitro diagnostic use.
## DESCRIPTION OF THE DEVICE:
The CALTAG CD3/CD19/CD45 monoclonal antibody combination binds to the surfaces of viable blood cells that express the corresponding antigens. To identify cells bearing these antigenic determinants, peripheral blood leukocytes are incubated with the monoclonal antibody, and washed to remove unbound antibody. Prior to removal of unbound antibody, lysis solution is added to lyse red blood cells. An appropriate fixative solution is added to lysed and washed cells. Stained and fixed cells are subsequently analyzed by flow cytometric methods.
## INTENDED USE OF THE DEVICE:
CALTAG CD3/CD19/CD45 is a fluorescent reagent containing a combination of CD3, CD19 and CD45 monoclonal antibodies conjugated to fluorescein, phycoerythrin and the tandem fluorochrome PE-Cy5, respectively. This reagent permits the simultaneous identification of CD3+ mature T lymphocytes, CD19+ mature B lymphocytes, and CD45+ leukocytes including lymphocytes, monocytes and granulocytes, by flow cytometric methods.
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SUMMARY OF THE TECHNICAL CHARACTERISTICS OF THE MANUFACTURER'S DEVICE COMPARED TO THE PREDICATE DEVICE:
Comparisons of Caltag Monoclonal Antibody Components to Caltag and Coulter Single Monoclonal Antibodies
| No. | Item | Caltag Components | Caltag Antibodies
Coulter Antibody | Comparison |
| --- | --- | --- | --- | --- |
| 1. | Intended Use | Flow Cytometry | Flow Cytometry
Immunofluorescence | Substantially equivalent |
| 2. | Specificity | CD3 | CD3 Caltag | Substantially equivalent |
| | | CD19 | CD19 Caltag | |
| | | CD19 | CD19 Coulter | |
| | | CD45 | | "live gating" only |
| 3. | Target cell
for CD3 | Mature | Mature | Substantially equivalent |
| | | T Lymphocyte | T Lymphocytes | |
| | for CD19 | B lymphocytes | B lymphocytes | Substantially equivalent |
| | for CD45 | All Leukocytes | All Leukocytes | "live gating" only |
| 4. | Chemical form | Monoclonal antibody | Monoclonal antibody | Substantially equivalent |
| 5. | Fluorochromes | FITC | FITC | Substantially equivalent |
| | | R-PE | R-PE (Caltag) | |
| | | R-PE | RD1 (Coulter) | |
| | | TRI-COLOR | not available (Coulter) | |
| 6. | Available forms | | | |
| | FITC | liquid, PBS | lyophilized | Substantially equivalent |
| | PE | liquid, PBS | liquid, PBS | |
| | TRI-COLOR | liquid, PBS | none available (Coulter) | |
| 7. | Sample prep.
methods | whole blood | whole blood | Substantially equivalent |
NON CLINICAL TESTS SUPPORTING A DETERMINATION OF SUBSTANTIAL EQUIVALENCE:
**EXPECTED VALUE DATA**
Blood samples were collected from a total of 155 apparently healthy normal donors in an age range of 16 to 72, with a mean age of 41, for the determination of expected values of the Caltag CD3/CD19/CD45 monoclonal antibody combination. Samples were collected and analyzed in each of three independent laboratories. An approximately equal number of males and females were collected and analyzed in each laboratory.
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The population contained members of differing ethnic origins, including adult Caucasians, Blacks, Orientals and Hispanics. Donors in geographically diverse areas of the United States, including the Eastern, South Central and Western regions, participated in this study.
Summary of expected values for CALTAG CD3 FITC CD19 R-PE and CD45 TRI-COLOR components for 155 normal donors:
| procedure | mean % positive | S.D. | Range ±2 S.D. |
| --- | --- | --- | --- |
| CD3 FITC component | 72.8 | 7.7 | 57-88 |
| CD19 R-PE component | 13.3 | 4.5 | 4-22 |
| CD45 TRI-COLOR component | 100.0 | 0.1 | 100-100 |
Comparison of CALTAG CD3 FITC, CD19 R-PE and CD45 TRI-COLOR components to the CALTAG CD3 FITC, CD19 R-PE and CD45 TRI-COLOR single antibodies. Blood samples were collected from different randomly selected populations of adult normal donors having similar age, gender and ethnic distributions for the determination of expected values for the Caltag CD3 FITC and CD45 TRI-COLOR single monoclonal antibodies, and from the same population of adult normal donors for the determination of expected values for Caltag CD19 R-PE single antibody.
### SINGLE ANTIBODIES
| procedure | mean % positive | S.D. | Range ±2 S.D. | n |
| --- | --- | --- | --- | --- |
| CD3 FITC | 71.8 | 6.9 | 58-86 | 130 |
| CD19 R-PE | 13.0 | 4.2 | 5-21 | 155 |
| CD45 TRI-COLOR | 99.0 | 0.9 | 97-100 | 40 |
Expected values for pediatrics and adolescents have not been determined.
The values obtained from normal individuals may vary from laboratory to laboratory; therefore, it is recommended that each laboratory establish its own normal range.
## SPECIFICITY DATA
Blood samples were obtained from healthy normal donors of Caucasian, Black, Hispanic and Oriental ethnic origins. Samples of each donor were stained with CALTAG CD3 FITC/CD19 R-PE/CD45 TRI-COLOR monoclonal antibody combination. Cells contained in the lymphocyte, monocyte and granulocyte regions were selected for analysis. Separate samples from the same donors were prepared for analysis of red blood cells and platelets and stained with each of the CALTAG monoclonal antibodies.
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| CD3 FITC Component | | | | | |
| --- | --- | --- | --- | --- | --- |
| Ethnic | Percent of Stained Cells | | | | |
| Origin | Lymph. | Mono. | Gran. | Plt. | RBC |
| Caucasian | 65.2 | 1.7 | 1.5 | 0.6 | 0.3 |
| Caucasian | 81.4 | 1.4 | 0.5 | 0.3 | 0.4 |
| Hispanic | 79.2 | 1.9 | 0.6 | 0.3 | 0.4 |
| Oriental | 81.2 | 1.3 | 0.9 | 0.4 | 0.2 |
| Black | 84.9 | 0.9 | 0.6 | 0.4 | 0.4 |
| Mean | 78.4 | 1.4 | 0.8 | 0.4 | 0.3 |
| ±1 S.D. | 7.6 | 0.4 | 0.4 | 0.1 | 0.1 |
| CD19 R-PE Component | | | | | |
| --- | --- | --- | --- | --- | --- |
| Ethnic | Percent of Stained Cells | | | | |
| Origin | Lymph. | Mono. | Gran. | Plt. | RBC |
| Caucasian | 18.1 | 1.3 | 0.5 | 0.4 | 0.2 |
| Caucasian | 12.9 | 0.6 | 0.4 | 0.2 | 0.2 |
| Hispanic | 13.4 | 0.5 | 0.6 | 0.2 | 0.6 |
| Oriental | 12.1 | 0.8 | 0.5 | 0.3 | 0.3 |
| Black | 12.7 | 0.9 | 0.3 | 0.4 | 0.2 |
| Mean | 13.8 | 0.8 | 0.5 | 0.3 | 0.3 |
| ±1 S.D. | 2.4 | 0.3 | 0.1 | 0.1 | 0.2 |
| CD45 TRI-COLOR Component | | | | | |
| --- | --- | --- | --- | --- | --- |
| Ethnic | Percent of Stained Cells | | | | |
| Origin | Lymph. | Mono. | Gran. | Plt. | RBC |
| Caucasian | 100.0 | 100.0 | 100.0 | 0.8 | 0.2 |
| Caucasian | 100.0 | 100.0 | 100.0 | 0.5 | 0.3 |
| Hispanic | 100.0 | 100.0 | 100.0 | 0.3 | 0.4 |
| Oriental | 100.0 | 100.0 | 100.0 | 0.4 | 0.3 |
| Black | 100.0 | 100.0 | 100.0 | 0.3 | 0.2 |
| Mean | 100.0 | 100.0 | 100.0 | 0.5 | 0.3 |
| ±1 S.D. | 0.0 | 0.0 | 0.0 | 0.2 | 0.1 |
Specific and/or nonspecific antibody Fc binding to monocytes in a patient sample can be excluded by proper gating on lymphocytes on the flow cytometer.
## CLINICAL TESTS SUPPORTING A DETERMINATION OF SUBSTANTIAL EQUIVALENCE:
### CORRELATION DATA
The Correlation study was performed on 175 donors, including 155 normal and 20 abnormal donors.
Comparison of the CALTAG CD3 FITC conjugated monoclonal antibody component of CD3 FITC/CD19 R-PE/CD45 TRI-COLOR with the CALTAG CD3 FITC conjugated single monoclonal antibody:
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| procedure | mean % positive | r² value | slope | Y intercept |
| --- | --- | --- | --- | --- |
| CD3 FITC component | 69.7 | 94.8 | 1.01 | 0.33 |
| CD3 FITC single | 68.3 | | | |
CD3 FITC component
Slope + 1.01
y intercept + 0.33
Linear regression y = 0.33 + 1.01x
Comparison of the CALTAG CD19 R-PE conjugated monoclonal antibody component of CD3 FITC/CD19 R-PE/CD45 TRI-COLOR with the Caltag CD19 R-PE conjugated single monoclonal antibody:
| procedure | mean % positive | r² value | slope | Y intercept |
| --- | --- | --- | --- | --- |
| CD19 R-PE component | 16.8 | 98.9 | 1.02 | 0.08 |
| CD19 R-PE single | 16.4 | | | |
CD19 R-PE component
Slope + 1.02
y intercept + 0.08
Linear regression y = 0.08 + 1.02x
Comparison of the CALTAG CD19 R-PE conjugated monoclonal antibody component of CD3 FITC/CD19 R-PE/CD45 TRI-COLOR with the Coulter CD19 RD1 conjugated single monoclonal antibody:
| procedure | mean % positive | r² value | slope | Y intercept |
| --- | --- | --- | --- | --- |
| CD19 R-PE component | 16.8 | 98.1 | 0.95 | 1.29 |
| Coulter CD19 RD1 | 16.2 | | | |
CD19 R-PE component
Slope + 0.95
y intercept + 1.29
Linear regression y = 1.29 + 0.95x
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