(194 days)
Monoclonal Mouse Anti-Human Leukocyte Common Antigen, CD45, FITC-Conjugated, Clone T29/33 Monocrollar Wouse First House Anti-Human Mouse Anti-Human Monocyte. CD14, RPE-Conjugated, Clone TUK4 (Anti-CD) 1/RPE, TÜK4) have been developed for use in flow cytometry. This reagent may be used to optimize the gating of lymphocytes when analyzing peripheral whole blood or peripheral blood mononuclear cell preparations. This reagent is one component of the suggested monoclonal antibody (MAb) combination for routine immunophenoryping of lymphocytes in peripheral blood.
Purified mouse anti-human CD45, Clone T29/33, conjugated with fluorescein isothiocyanate, isomer 1 (FITC) + purified mouse anti-human CD14, Clone TUK4, 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.
Here's an analysis of the provided text, outlining the acceptance criteria and study details as requested:
Acceptance Criteria and Device Performance
| Acceptance Criteria | Reported Device Performance |
|---|---|
| Linearity of Anti-CD45/FITC: | y = 0.11% + 0.997x. r = 0.999 (for Anti-CD45/FITC, using Raji and U937 cells) |
| Reproducibility (Anti-CD45/FITC, %CV): | FACScan: 0.1%, 0.85%, 0.04% Profile II: 1.31%, 1.24%, 1.55% |
| Reproducibility (Anti-CD14/RPE, %CV): | FACScan: 21.92%, 15.46%, 18.63% Profile II: 8.66%, 15.58%, 6.60% |
| Specificity (CD45+CD14- for WBCs, not RBCs/platelets, Anti-CD45/FITC): | Avg % Positive Red Blood Cells: 0.1% (0.0-0.2%) Avg % Positive Granulocytes: 13.4% (1.9-35.4%) Avg % Positive Monocytes: 96.6% (93.7-98.8%) Avg % Positive Lymphocytes: 0.2% (0.0-0.3%) Avg % Positive Platelets: 0.1% (0.0-0.2%) The text indicates "antibody binding of Anti-CD45 T29/33 is specific for WBC's, not RBC's or platelets." |
| Specificity (CD14+ for monocytes, Anti-CD14/RPE): | The text states "Anti-CD14/RPE, TÜK4 is specific for monocytes, with some binding noted for granulocytes." (Numerical data from the table above also supports monocyte specificity with high % Positive Monocytes for CD45+CD14: and CD45-CD14+ blood cells, and some percentage for granulocytes) |
| Correlation to Predicate Device (Linearity for CD45+ cells): | y = 18.68 + 0.81 X (Predicate CD45+ cells). r² = 0.8847 |
| Correlation to Predicate Device (Linearity for CD14+ cells): | y = 0.44 + 0.91 X (Predicate CD14+ cells). r² = 0.6187 |
| Gating Verification (Lymphocyte population): | 99% or more of the cells as CD45+ CD14 as 2% or less of the cell population. |
Study Details
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Sample size used for the test set and the data provenance:
- Linearity (Anti-CD45/FITC): Not explicitly stated how many individual cells/samples were used, but it involved serial dilutions of Raji (antigenic) and U937 (non-antigenic) cell lines.
- Reproducibility: 10 replicates from peripheral blood of one donor per test (for each of two flow cytometers and each antibody). Data provenance is DAKO Corporation internal testing.
- Specificity: 5 apparently healthy adult donors (3 Caucasians, 1 Asian, 1 Hispanic) for basic specificity. Data provenance is DAKO Corporation internal testing.
- Correlation to Predicate Device: 150 normal, apparently healthy individuals and 27 samples from ill patients. These were tested at three geographically separate laboratories. Data provenance is multi-site, multi-ethnic (implied by "normal, apparently healthy individuals"), and appears to be prospective for the purpose of this comparison.
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Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- The document does not mention experts being used to establish ground truth for the specific performance evaluation (reproducibility, specificity, linearity). These appear to be laboratory measurements against known cell lines or direct observation of antibody binding.
- For the correlation study against the predicate device, the "ground truth" is essentially the results obtained from the predicate device itself. The document does not specify if experts verified these predicate results.
- The "Leukocyte Typing Workshops" (Third, Fourth, Fifth) are cited as providing information that contributes to the conclusions, implying a broader expert consensus within the scientific community for the validity of the clones used (T29/33 and TUK4). However, these workshops did not establish the ground truth for this specific device's test set.
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Adjudication method for the test set:
- No adjudication method is described for the direct performance studies (linearity, reproducibility, specificity).
- For the correlation study, duplicate samples were tested with each reagent, suggesting comparative measurement rather than adjudication by experts.
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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:
- No MRMC comparative effectiveness study was done. This device is an in vitro diagnostic reagent and accessory for flow cytometry, not an AI or imaging device where "human readers" typically interpret results with or without assistance in the conventional sense. The "reading" is done by the flow cytometer, and the results are numerical.
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If a standalone (i.e., algorithm only without human-in-the-loop performance) was done:
- This is not applicable as the device is a reagent for flow cytometry, not an algorithm. The flow cytometer itself performs the "standalone" measurement, and human operators perform sample preparation, instrument setup, and interpretation of the numerical results produced by the instrument. The "algorithm" here would be the flow cytometer's software, which is part of the system the reagent is designed to be used with.
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The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- Linearity: Based on known antigenic expression of established cell lines (Raji and U937).
- Reproducibility: Based on direct measurements from a single donor's peripheral blood run multiple times.
- Specificity: Based on observed antibody binding to different cell populations (RBCs, granulocytes, monocytes, lymphocytes, platelets) in apparently healthy donors, with cell identification likely based on morphology and characteristic light scatter patterns in flow cytometry, consistent with cellular biology.
- Correlation to Predicate Device: The "ground truth" for the comparative study was the results obtained from the predicate device (Becton Dickinson Simultest LeucoGATE) on the same samples. These predicate results are assumed to be a valid measurement.
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The sample size for the training set:
- This is not applicable. The device is a diagnostic reagent, not a machine learning algorithm that requires a "training set." The development of the monoclonal antibodies (clones T29/33 and TUK4) themselves would have involved extensive R&D, but that is not an "algorithm training set" in the modern sense.
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How the ground truth for the training set was established:
- Not applicable, as there is no "training set" for this type of device.
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JUN 24 1991
સ્વર્જ્યનયત્વે
| Submitter: | DAKO Corporation6392 Via RealCarpinteria, CA 93013(805)566-6655 | ||
|---|---|---|---|
| Contact: | Gretchen M. Murray, Ph.D., Regulatory Affairs Asst. Manager | ||
| Date SummaryPrepared: | June 19, 1997 | ||
| Device Name: | Mouse Anti-Human CD45/FITC, T29/33 +Mouse Anti-Human CD14/RPE, TUK4 | ||
| DeviceClassification: | Class II according to 21 CFR 864.5220, on the basis that monoclonalantibodies 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 LeucoGATE | ||
| Device Description: | Purified mouse anti-human CD45, Clone T29/33, conjugated with fluoresceinisothiocyanate, isomer 1 (FITC) + purified mouse anti-human CD14, CloneTUK4, conjugated with R-phycoerythrin, present in 0.05M Tris-HCl buffer, pH7.2, 15 mM NaN3, 0.1M NaCl, stabilized with 1% carrier protein | ||
| Subpopulations of lymphocytes may be stained with fluorochrome-conjugatedantibody and evaluated in peripheral blood specimens when contaminating redblood cells (RBC's) are lysed prior to flow cytometric analysis. A subpopulationof WBC's are selected for assessment based upon cell morphology. | |||
| Intended Use: | For In Vitro Diagnostic Use | ||
| Mouse Anti-Human CD45/FITC, T29/33 + Mouse Anti-Human CD14/RPE,TUK4 (DAKO Anti-CD45/FITC and Anti-CD14/RPE) has been developed for usein flow cytometry to optimize the gating of lymphocytes when analyzingperipheral whole blood (erythrocyte lysed peripheral blood samples) orperipheral blood mononuclear cell preparations. It is one component of thesuggested monoclonal antibody (MAb) combinations for routineimmunophenotyping of lymphocytes in peripheral blood using flow cytometry. | |||
| Comparison ofTechnologicalCharacteristics | Anti-CD45/FITC, T29/33, and Anti-CD14/RPE, TUK4 is a dual antibody reagentin which each antibody has a related fluorescent conjugate. This reagent isused for detection and enumeration in peripheral blood. Clone T29/33 wasclustered at the Third and Fourth International Leukocyte Workshops (In:McMichael, AJ, et al (Ed's). Leukocyte Typing III, Oxford, New York, Tokyo:Oxford University Press, 1987). Clone TUK4 was clustered at the FourthInternational Leukocyte Workshop (Knapp, W, et al (eds). Leukocyte TypingIV, Oxford, New York, Tokyo: Oxford University Press, 1989.) | ||
| Binding linearity was determined over serial dilutions of a cell line known to |
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express the antigen diluted with a cell line that has no antigenic sites for each antibody. For Anti-CD45/FITC, the cell line with known antigenic reactivity is Raji cells, while the cell line without antigenic sites is U937 cells. For Anti-CD14/RPE, no cell line could be reliably grown for linearity testing. Linearity results are not available for this antibody. For Anti-CD45/FITC, five dilutions were rested, with linear equations calculated from the results. The equation for Anti-CD45/FITC, T29/33 was y = 0.11% + 0.997x. r = 0.999.
Reproducibility of ten replicates from peripheral blood of one donor were run on two flow cytometers from different manufacturers. Different concentrations of antigen from whole blood was not possible to obtain.
| Anti-CD45/FITC | Mean % CD45+ | ± 1 SD | %CV | n |
|---|---|---|---|---|
| FACScan | 99.8 | 0.1 | 0.1 | 10 |
| 99.7 | 0.84 | 0.85 | 10 | |
| 100 | 0.04 | 0.04 | 10 | |
| Profile II | 88.58 | 1.16 | 1.31 | 10 |
| 95.61 | 1.18 | 1.24 | 10 | |
| 94.65 | 1.47 | 1.55 | 10 | |
| Anti-CD14/RPE | Mean % CD14+ | ± 1 SD | %CV | n |
| FACScan | 2.30 | 0.50 | 21.92 | 10 |
| 2.63 | 0.41 | 15.46 | 10 | |
| 1.57 | 0.29 | 18.63 | 10 | |
| Profile II | 3.02 | 0.26 | 8.66 | 10 |
| 3.67 | 0.57 | 15.58 | 10 | |
| 6.41 | 0.42 | 6.60 | 10 |
Specificity of Anti-CD45/FITC, T29/33 in combination with Anti-CD14/RPE, TÜK4 has been verified by tests performed on five apparently healthy adult donors of various races at DAKO Corporation (3 Caucasians, 1 Asian, 1 Hisbanic). Cell populations tested were RBC's, granulocytes, monocytes, lymphocytes and platelets. The results indicate antibody binding of Anti-CD45 T29/33 is specific for WBC's, not RBC's or platelets. Anti-CD14/RPE, TÜK4 is specific for monocytes, with some binding noted for granulocytes.
| Averages(n = 5) | % PositiveRed BloodCells | % PositiveGranulocytes | % PositiveMonocytes | % PositiveLymphocytes | %PositivePlatelets | |
|---|---|---|---|---|---|---|
| CD45+CD14-Blood Cells | 0.1(0.0-0.2) | 13.4(1.9-35.4) | 96.6(93.7-98.8) | 0.2(0.0-0.3) | 0.1(0.0-0.2) | |
| CD45+CD14:Blood Cells | 0.4(0.1-1.1) | 99.5(98.0-100.0) | 99.3(97.1-100.0) | 99.0(96.9-99.9) | 2.0(0.4-4.5) | |
| CD45-CD14+Blood Cells | 0.2(0.0-0.5) | 13.5(1.9-35.4) | 96.6(93.7-98.8) | 0.3(0.0-0.5) | 0.7(0.4-1.2) |
Anti-CD45/FITC with Anti-CD14/RPE Specificity
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Correlation of the dual antibody reagent, Anti-CD45/FITC, T29/33, and Anti-CD14/RPE, TUK4 to a predicate dual antibody reagent, Becton Dickinson Simultest LeucoGATE, was determined by testing duplicate samples with each reagent across 150 normal, apparently healthy individuals at three geographically separate laboratories and 27 samples obtained from ill patients. The regression analysis indicated that there was a 1:1 linear comparison of the DAKO CD45/CD14 to the LeucoGATE CD45/CD14 CD45/FITC. The linear equations that were generated are:
VIDAKO CO45/CD16 CO45 F callaj = 18.68 + 0.81 X(LoudaGATE CD45 + MIN). r2 = 0.8847
(DAKO CD45/CD14 CD14 .; colla) = 0.44 + 0.91X(1,BUcaGATE CD14 .; calle) . r2 = 0.6187
These equations indicate that the Anti-CD45/FITC, T29/33, and Anti-CD14/RPE, TUK4 reagent and the Becton Dickinson Simultest LeucoGATE reagent are comparable on a 1:1 basis.
Conclusions:
Results of the above testing as well as the information provided by the Third, Fourth and Fifth Leukocyte Typing Workshops indicate that the DAKO Anti-CD45/FITC plus Anti-CD14/RPE reagent performs as well as Becton Dickinson's Simultest LeucoGATE in the detection and enumeration of CD45 ' lymphocytes and the labeling of CD14 positive cells for the exclusion of these cells using flow cytometry for the detection and enumeration of lymphocytes. Safety of the DAKO Anti-CD45/FITC plus Anti-CD14/RPE reagent and its predicate device is high as all reagents are used for in vitro testing.
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Image /page/3/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo is a circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract symbol that resembles a stylized caduceus, a symbol often associated with medicine and healthcare. The symbol consists of a staff with a serpent entwined around it, and a pair of wings at the top.
Food and Drug Administration 2098 Gaither Road Rockville MD 20850
JUN 24 1997
Gretchen M. Murray, Ph.D. Regulatory Affairs Asst. Manager DAKO Corporation 6392 Via Real ………… Carpinteria, CA 93013
Re: K964974/S001
Trade Name: Monoclonal Mouse Anti-Human Leukocyte Common Antigen, CD45, FITCconjugated. Clone T29/33 and Monoclonal Mouse Anti-Human Monocyte, CD14, RPE Conjugated, Clone TOK 4 Regulatory Class: II Product Code: GKZ Dated: March 28, 1997 Received: April 04, 1997
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 regulations 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 Good Manufacturing Practice for Medical Devices: General (GMP) regulation (21 CFR Part 820) and that, through periodic GMP 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 Register.
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.
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.
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Page 2
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 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 Autman
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 _ of _
0(k) Number (if known):
evice Name: Monoclonal Mouse Anti-Human Leucocyte Common Antigen, CD45, FITCevice Name: monocronal nouse internant used on t-Human Monocyte, CD14, RPE-Conjugated, Clone TUK4 dications For Use:
Monoclonal Mouse Anti-Human Leukocyte Common Antigen, CD45, FITC-Conjugated, Clone T29/33 Monocrollar Wouse First House Anti-Human Mouse Anti-Human Monocyte. CD14, RPE-Conjugated, Clone TUK4 (Anti-CD) 1/RPE, TÜK4) have been developed for use in flow cytometry. This reagent may be used to optimize the gating of lymphocytes when analyzing peripheral whole blood or peripheral blood mononuclear cell preparations. This reagent is one component of the suggested monoclonal antibody (MAb) combination for routine immunophenoryping of lymphocytes in peripheral blood.
Lymphocyte gating is assigned by selection of the upper and lower channel numbers to define the Ivmphocye population and is a quality control function for flow cytometry. Gating verification is lymplocere population and 16 9911% of the cells as CD45 meth CD14 as 2% or less of the cell population. 19
Note that this definition of "lymphocyte" does not give complete and unambiguous resolution from all other cell types. When optimizing light scatter gates for lymphocytes, operator and/or software algorithms are minimizing the number of lymphocytes excluded while still maintaining acceptable levels of contaminating cell types. Lymphocyte gate purity may be calculated by determining the percentage of non-lymphocyte events within the gate. Most non-lymphocytes are CD14 positive. Because each flow cytometer has different operating characteristics, each laboratory must determine its optimal operating procedure.
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Division of Clinical Laborator tory Devices
510(k) Number
Citeur Matin
(PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON ANOTHER PAGE IF NEEDED)
Concurrence of CDRH, Office of Device Evaluation (ODE)
escription Use ✓
er 21 CFR 801.109)
D Use
er 21 CFR 801.119)
OR
Over-The-Counter Use
(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.”