(153 days)
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.
Monoclonal Mouse Anti-Human B-cell, CD19, RPE-Cy5-conjugated, Clone HD37 is specific for B-lymphocyte cluster determinants as evaluated by the 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.
This document describes the DAKO® Mouse Anti-Human B-cell, CD19/RPE-Cy5, Clone HD37 (Product Code No. C7066), a monoclonal antibody reagent intended for flow cytometry analysis of B-cells in peripheral blood.
Acceptance Criteria and Device Performance Study Summary
The acceptance criteria for this device are not explicitly stated in a quantitative manner as "acceptance criteria" but are implied through the performance characteristics presented. The study demonstrates the device's linearity, reproducibility, specificity, and correlation with a predicate device.
1. Table of Acceptance Criteria and Reported Device Performance
| Performance Characteristic | Acceptance Criteria (Implied) | Reported Device Performance |
|---|---|---|
| Binding Linearity | $R^2$ value close to 1 for the linear fit of antigen dilution vs. signal. | $y = 0.01% + 0.98x$, $R^2 = 0.999$ (for Anti-CD19/Cy5, HD37) |
| Reproducibility | Low Coefficient of Variation (%CV) across different concentrations and machines. | FACScan: High Level (90.87 ± 1.24, 1.37%CV); Medium Level (51.28 ± 1.47, 2.87%CV); Low Level (26.77 ± 1.02, 3.81%CV).Profile II: High Level (87.16 ± 2.09, 2.40%CV); Medium Level (49.75 ± 1.54, 3.09%CV); Low Level (26.13 ± 1.08, 4.11%CV). |
| Specificity | Minimal non-specific binding, particularly to non-lymphocyte populations; specific binding to B-cells (lymphocytes). | Specific for lymphocytes (average 13.0% binding, representative of B-cell population). Minimal binding to RBCs (0.04%), granulocytes (1.10%), and platelets (0.22%). Some binding to monocytes (7.66%), but can be excluded by proper gating. |
| Predicate Equivalence (Normal) | Strong linear correlation ($R^2$ close to 1) with the predicate device. | $y_{\text{(DAKO CD19/Cy5+ Lymphocytes)}} = 2.27 + 0.77 x_{\text{(DAKO CD19/RPE + Lymphocytes)}}$. $R^2 = 0.6743$. |
| Predicate Equivalence (Total) | Strong linear correlation ($R^2$ close to 1) with the predicate device across a broader range of samples, ideally a 1:1 relationship (slope near 1, intercept near 0). | $y_{\text{(DAKO CD19/Cy5+ Lymphocytes)}} = 0.45 + 0.98 x_{\text{(DAKO CD19/RPE + Lymphocytes)}}$. $R^2 = 0.8832$. This indicates comparability on a 1:1 basis (slope 0.98, intercept 0.45). |
2. Sample Size Used for the Test Set and Data Provenance
- Linearity: 5 dilutions of Raji cells (antigen-positive) diluted with JM cells (antigen-negative). Data provenance is internal (DAKO Corporation, implicitly).
- Reproducibility: 10 replicates from peripheral blood of 3 donors. Data provenance is internal (DAKO Corporation, implicitly).
- Specificity: 5 apparently healthy adult donors of various races. Data provenance is internal (DAKO Corporation, implicitly).
- Predicate Equivalence:
- 153 normal, apparently healthy individuals. Data provenance: prospective, from three geographically separate laboratories.
- An additional 27 samples from patients with illnesses. Data provenance: not explicitly stated if prospective or retrospective, but added to the overall dataset for correlation.
- Total for correlation: 180 samples.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of those Experts
The document does not mention the use of experts to establish a "ground truth" in the traditional sense for the performance studies. The measurements (e.g., %CD19+ cells) are direct measurements from flow cytometry, which is an objective measurement technique rather than interpretive.
4. Adjudication Method for the Test Set
Not applicable. The measurements are objective laboratory results and do not require adjudication.
5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
Not applicable. This device is a monoclonal antibody reagent used in flow cytometry, not an AI or imaging diagnostic tool that involves human "readers" or subjective interpretation.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
This refers to the performance of the device itself (the antibody reagent) in binding to target cells and being detected by a flow cytometer. The entire testing described (linearity, reproducibility, specificity, predicate comparison) represents the standalone performance of the reagent. Human involvement is in operating the flow cytometer and analyzing the raw data, but the performance metrics directly pertain to the reagent's characteristics.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.)
The "ground truth" for the performance studies is rooted in the intrinsic properties of the cells and the flow cytometry method itself:
- Linearity: Based on known concentrations of antigen-expressing cells.
- Reproducibility: Based on repeated measurements of the same samples.
- Specificity: Verified by testing binding to different known cell populations (RBCs, granulocytes, monocytes, lymphocytes, platelets) and observing the expected pattern of strong binding to lymphocytes (B-cells) and minimal binding to others.
- Predicate Equivalence: The predicate device (DAKO Anti-CD19/RPE, HD37 reagent) serves as the "reference standard" or "ground truth" for comparative purposes, implicitly having its own established performance.
8. The sample size for the training set
The document does not mention a "training set" in the context of machine learning or AI algorithms. This device is a biochemical reagent, and its development would typically involve empirical optimization and validation rather than a formal machine learning training phase.
9. How the ground truth for the training set was established
Not applicable, as there is no "training set" in the context of the type of device described.
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.111 22 1998
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 RedBlood Cells | % PositiveGranulocytes | % PositiveMonocytes | % PositiveLymphocytes | % PositivePlatelets | |
|---|---|---|---|---|---|
| Average (n=5)(range) | 0.04(0.0-0.2) | 1.10(0.3-1.7) | 7.66(3.5-9.7) | 13.34(10.1-17.6) | 0.22(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:
$y_{\text{(DAKO CD19/Cy5+ Lymphocytes)}} = 0.45 + 0.98 x_{\text{(DAKO CD19/RPE + Lymphocytes)}}$.
6
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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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Page 2
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 (770)488-7655.
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 (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. 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 (800) 638-2041 or at (301) 443-6597 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)
§ 864.5220 Automated differential cell counter.
(a)
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.(b)
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.”