CALTAG CD19 R-PE and CD19 TRI-COLOR are fluorochrome conjugated monoclonal antibody reagents that may be used to enumerate CD19+ lymphocytes in human peripheral blood by flow cytometric methods.

Device Description

The CALTAG CD19 R-PE and CD19 TRI-COLOR monoclonal antibodies bind to the surfaces of viable blood cells that express the CD19 antigen. To identify cells bearing the CD19 determinant, 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.

AI/ML Overview

Here's a breakdown of the acceptance criteria and study information for the CALTAG CD19 R-PE and CD19 TRI-COLOR Mouse Monoclonal Antibodies, based on the provided text:

Acceptance Criteria and Reported Device Performance

The provided document primarily focuses on establishing substantial equivalence to predicate devices rather than defining explicit pass/fail acceptance criteria. However, we can infer some criteria from the comparisons made and the expected values presented. The "Comparison" column in the table implies the acceptance criterion of "Substantially equivalent" to the predicate devices for each item.

No.	Item	Acceptance Criteria (Inferred)	Reported Device Performance (CALTAG Antibodies)	Predicate Device Performance (Coulter Antibodies)	Comparison Result
1.	Intended Use	Substantially equivalent to predicate	Flow Cytometry	Flow Cytometry, Immunofluorescence	Substantially equivalent
2.	Specificity	Substantially equivalent to predicate	CD19	CD19	Substantially equivalent
3.	Target cell	Substantially equivalent to predicate	B lymphocyte	B lymphocyte	Substantially equivalent
4.	Chemical form	Substantially equivalent to predicate	Monoclonal antibody	Monoclonal antibody	Substantially equivalent
5.	Fluorochromes	Substantially equivalent to predicate	R-PE, TRI-COLOR	FITC, RD1	Substantially equivalent
6.	Available forms	Substantially equivalent to predicate	FITC liquid, PBS; PE liquid, PBS; TRI-COLOR liquid, PBS	lyophilized, liquid, PBS, not available	Substantially equivalent
7.	Sample prep. methods	Substantially equivalent to predicate	whole blood	whole blood	Substantially equivalent
8.	Expected values from this study (n=155)	Within or comparable to predicate device's expected range (4-21% for RD1, 3-23% for FITC)	R-PE 5-21%, TRI-COLOR 4-24%	4-21% (RD1), 3-23% (FITC)	Substantially equivalent

Reproducibility (Intra-lab and Inter-lab): While specific acceptance criteria for %CV are not explicitly stated, the data provided implies that the observed %CV values in various ranges (high, mid, low) and across different sites were considered acceptable for demonstrating reproducibility. The ranges of observed %CV were:

CD19 R-PE (Intra-lab): 0.5% - 7.6%
CD19 TRI-COLOR (Intra-lab): 1.0% - 6.0%

Correlation: For correlation studies, the acceptance criteria are implicitly that the R-squared (r²) value, slope, and Y-intercept indicate strong linear correlation and agreement with the predicate devices. The reported r² values are very high:

CALTAG CD19 R-PE vs. Coulter CD19 RD1: r² = 97.7
CALTAG CD19 R-PE vs. Coulter CD19 FITC: r² = 96.3
CALTAG CD19 TRI-COLOR vs. Coulter CD19 RD1: r² = 96.7
CALTAG CD19 TRI-COLOR vs. Coulter CD19 FITC: r² = 97.4
CALTAG CD19 TRI-COLOR vs. CALTAG CD19 R-PE: r² = 97.6

The slopes are close to 1 (0.92 to 0.99) and Y-intercepts are close to 0 (-0.56 to 2.14), indicating strong agreement.

Study Information

Sample size used for the test set and the data provenance:
- Expected Value Data: 155 apparently healthy normal donors.
- Specificity Data: An unspecified number of healthy normal donors (Caucasian, Black, Hispanic and Oriental ethnic origins). The table shows data for 5 different donor samples.
- Reproducibility Data (Intra-lab): For each antibody and each of three ranges (high, medium, low), 10 replicated determinations were performed. This involved 10 separate samples per range. The study was conducted in each of three independent laboratories.
- Reproducibility Data (Inter-lab): Similar to intra-lab, 10 replicated determinations for each antibody in each of three ranges (high, medium, low). This involved blood samples from "the same blood donors" across three laboratories.
- Correlation Data: 175 donors (155 normal and 20 abnormal donors).
- Data Provenance: Geographically diverse areas of the United States, including Western, Eastern, and SouthCentral regions. The studies appear to be prospective, as blood samples were "collected" and "analyzed" for the purpose of this study.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts: Not applicable. This study focuses on the performance of monoclonal antibodies for flow cytometry, a lab assay. The "ground truth" is typically established by the assay itself and comparison to established methods (predicate devices), not by expert interpretation of images or other data.
Adjudication method (e.g., 2+1, 3+1, none) for the test set: Not applicable. This is a laboratory assay where results are quantifiable percentages, not subjective interpretations requiring adjudication.
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 study is about the performance of monoclonal antibodies, not an AI or human reader effectiveness study.
If a standalone (i.e., algorithm only without human-in-the loop performance) was done: The devices are reagents for flow cytometry, which is an automated process for cell counting and phenotyping. The performance presented is of the reagents in conjunction with standard flow cytometric methods, which can be seen as an 'algorithm only' performance in terms of the reagent's interaction with the biological sample and the instrument, without human interpretation as the primary output.
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- Expected Values and Specificity: The "ground truth" for the expected values and specificity is derived from flow cytometric analysis of samples from clearly defined populations (healthy donors, specific cell types) using the test device and comparing it to the known performance of predicate devices.
- Reproducibility: The ground truth for reproducibility is the consistency of the assay results itself, measured by standard deviation (S.D.) and coefficient of variation (%CV).
- Correlation: The ground truth for correlation is the established performance of the legally marketed predicate devices (Coulter CD19 RD1 and CD19 FITC monoclonal antibodies), against which the new CALTAG antibodies are compared using linear regression.
The sample size for the training set: Not applicable. This is a medical device study for reagents, not an AI model requiring a training set. The term "training set" is typically used in machine learning.
How the ground truth for the training set was established: Not applicable, as there is no training set for an AI model.

Summary

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Image /page/0/Picture/0 description: The image shows the logo for CALTAG LABORATORIES. The logo is in black and white, with the word "CALTAG" in large, bold letters on top. Below "CALTAG" is a horizontal line, and below the line is the word "LABORATORIES" in smaller, but still bold, letters. The logo is simple and professional.

K963954

510(K) SUMMARY SUMMARY OF SAFETY AND EFFECTIVENESS DATA

NOV 25 1996

CD19 R-PE, CD19 TRI-COLOR Mouse Monoclonal Antibodies 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:

October 1, 1996

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TRADE NAME OF THE DEVICE:

Caltag CD19 R-PE, CD19 TRI-COLOR Mouse Monoclonal Antibodies To Human Cell Surface Antigens by Flow Cytometry

COMMON NAME:

Caltag CD19 R-PE, CD19 TRI-COLOR Monoclonal Antibody

CLASSIFICATION NAME:

Automated Differential Cell Coulter (21 CFR 864.5220)

LEGALLY MARKETED DEVICE (PREDICATE DEVICE) TO WHICH THE MANUFACTURER IS CLAIMING SUBSTANTIAL EQUIVALENCE:

Caltag CD19 R-PE Monoclonal Antibody to Human Cell Surface Antigens is substantially equivalent to the Coulter CD19 RD1 Monoclonal antibody for invitro diagnostic use.

Caltag CD19 TRI-COLOR Monoclonal Antibody to Human Cell Surface Antigens is substantially equivalent to the Coulter CD19 FITC and CD19 RD1 monoclonal antibodies for in-vitro diagnostic use.

DESCRIPTION OF THE DEVICE:

INTENDED USE OF THE DEVICE:

CALTAG CD19 R-PE and CD19 TRI-COLOR are fluorochrome conjugated monoclonal antibody reagents that may be used to enumerate CD19+ lymphocytes in human peripheral blood by flow cytometric methods.

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SUMMARY OF THE TECHNICAL CHARACTERISTICS OF THE MANUFACTURER'S DEVICE COMPARED TO THE PREDICATE DEVICE:

No.	Item	Caltag Antibodies	Coulter Antibodies	Comparison
1.	Intended Use	Flow Cytometry	Flow CytometryImmunofluorescence	Substantiallyequivalent
2.	Specificity	CD19	CD19	Substantiallyequivalent
3.	Target cell	B lymphocyte	B lymphocyte	Substantiallyequivalent
4.	Chemical form	Monoclonalantibody	Monoclonalantibody	Substantiallyequivalent
5.	Fluorochromes	R-PE,TRI-COLOR	FITC,RD1	Substantiallyequivalent
6.	Available forms	FITC liquid, PBSPE liquid, PBSTRI-COLOR liquid, PBS	lyophilizedliquid, PBSnot available	Substantiallyequivalent
7.	Sample prep.methods	whole blood	whole blood	Substantiallyequivalent
8.	Expected valuesfrom this study (n=155)	R-PE 5-21%TRI-COLOR 4-24%	4-21% (RD1)3-23% (FITC)	Substantiallyequivalent

Comparisons of Caltag CD19 and Coulter CD19 Monoclonal Antibodies

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. 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.

The normal donor population included members of differing ethnic origins, including adult Caucasian, Black, Oriental and Hispanic.

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Donors in geographically diverse areas of the United States, including the Western, Eastern and SouthCentral regions, participated in this study. Blood samples collected from each donor were stained with the CALTAG CD19 R-PE and CD19 TRI-COLOR monoclonal antibodies.

Summary of expected values for CALTAG CD19 monoclonal antibodies for all normal donors:

procedure	mean% positive	S.D.$\pm$2 S.D.	Range	n
CD19 R-PE	13.0	4.2	5-21	155
CD19 TRI-COLOR	13.9	5.0	4-24	155

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 CD19 R-PE and CD19 TRI-COLOR monoclonal antibodies. 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.

CD19 R-PE

EthnicOrigin	Lymph.	Mono.	Gran.	Plt.	RBC
Caucasian	18.0	0.6	0.9	0.5	0.5
Caucasian	13.3	1.1	0.8	0.3	0.7
Hispanic	12.2	0.7	0.8	0.4	1.0
Oriental	11.2	1.6	1.3	0.4	0.5
Black	14.6	0.0	0.5	0.6	0.9
Mean	13.9	0.8	0.9	0.4	0.7
± 1 S.D.	2.6	0.6	0.3	0.1	0.2

CD19 TRI-COLOR

EthnicOrigin	Lymph.	Mono.	Gran.	Plt.	RBC
Caucasian	18.3	0.0	1.0	0.3	0.4
Caucasian	12.1	0.2	1.1	0.4	0.1
Hispanic	11.6	0.2	0.3	0.2	0.7
Oriental	11.0	1.0	0.9	0.3	0.2
Black	12.0	0.7	0.9	0.4	0.2
Mean	13.0	0.4	0.8	0.3	0.3
$\pm$ 1 S.D.	3.0	0.4	0.3	0.1	0.2

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.

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REPRODUCIBILITY DATA (INTRA-LAB)

Intra-lab reproducibility for the CALTAG CD19 R-PE and CD19 TRI-COLOR conjugated monoclonal antibodies was determined by performing 10 replicated determinations for each antibody in each of three ranges; high, medium and low. Thus, a total of 30 determinations were performed for each form of CD19. In this manner, reproducibility was demonstrated throughout the entire measuring range.

The 10 determinations for each range were performed by the staining, processing and analysis of 10 separate samples. Lymphocytes were selected for the analysis of percent cells stained in each of the three ranges.

To perform this study, anticoagulated blood was obtained from an abnormal donor expressing a high percentage of CD19+ cells. Mid range and low range samples were obtained by adding known CD19- cells in appropriate ratios, while maintaining approximately the same total cell concentrations for the three ranges.

The study was performed in each of three independent laboratories, in the manner that each laboratory obtained, stained and analyzed separate blood samples. The following data are representative:

procedure	Level	mean% positive	S.D.	% CV	n
CD19 R-PE	high	66.6	0.4	0.5	10
	mid	46.7	0.8	1.6	10
	low	14.9	0.6	4.3	10
procedure	Level	mean% positive	S.D.	% CV	n
CD19TRI-COLOR	high	65.4	0.7	1.0	10
	mid	46.1	0.6	1.3	10
	low	15.5	0.4	2.7	10
			-6-
SITE 1
procedure	Level	mean% positive	S.D.	% CV	n
CD19 R-PE	high	84.7	4.4	5.2	10
	mid	70.4	1.2	1.8	10
	low	42.7	3.2	7.6	10
procedure	Level	mean% positive	S.D.	% CV	n
CD19TRI-COLOR	high	86.7	1.0	1.2	10
	mid	70.9	1.3	1.9	10
	low	42.7	0.9	2.1	10
SITE 2
procedure	Level	mean% positive	S.D.	% CV	n
CD19 R-PE	high	83.9	1.5	1.7	10
	mid	71.1	2.8	3.9	10
	low	42.5	1.9	4.6	10
procedure	Level	mean% positive	S.D.	% CV	n
CD19TRI-COLOR	high	85.3	2.2	2.6	11
	mid	65.7	3.1	4.8	9
	low	32.6	2.0	6.0	10
SITE 3
procedure	Level	mean% positive	S.D.	% CV	n
CD19 R-PE	high	87.5	0.6	0.7	10
	mid	69.9	0.8	1.1	10
	low	38.8	1.6	4.2	10
procedure	Level	mean% positive	S.D.	% CV	n
CD19TRI-COLOR	high	85.3	0.9	1.0	10
	mid	66.1	1.0	1.5	10
	low	30.7	2.4	7.9	10

REPRODUCIBILITY, (INTER-LAB)

Inter-lab reproducibility for the CALTAG CD19 R-PE and CD19 TRI-COLOR conjugated monoclonal antibodies was determined by performing 10 replicated determinations for each antibody in each of three ranges; high, medium and low. Thus, a total of 30 determinations were performed for each form of CD19. In this manner, reproducibility was demonstrated throughout the entire measuring range.

The study was performed in each of three laboratories. All laboratories stained and analyzed blood samples from the same blood donors. Lysed unstained samples containing cells in the appropriate ranges were prepared by one of the participating laboratories for staining and analysis in each of the laboratories. The following data were obtained:

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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 CD19 R-PE conjugated monoclonal antibody with the Coulter CD19 RD1 conjugated monoclonal antibody:

procedure	mean %positive	r2value	slope	Yintercept	n
CD19 R-PE	16.4	97.7	0.92	1.30	175
CD19 RD1	16.2
CD19 R-PE

y = 1.30 + 0.92x Linear regression

Comparison of the CALTAG CD19 R-PE conjugated monoclonal antibody with the Coulter CD19 FITC conjugated monoclonal antibody:

procedure	mean %positive	r2value	slope	Yintercept	n
CD19 R-PE	16.4	96.3	0.93	0.69	175
CD19 FITC	16.7

CD19 R-PE

Linear regression y = 0.69 + 0.93x

Comparison of the CALTAG CD19 TRI-COLOR conjugated monoclonal antibody with the Coulter CD19 RD1 conjugated monoclonal antibody:

procedure	mean %positive	r²value	slope	Yintercept	n
CD19 TRI-COLOR	17.1	96.7	0.92	2.14	175
CD19 RD1	16.2
CD19 TRI-COLORLinear regression	$y =2.14 + 0.92x$

Comparison of the CALTAG CD19 TRI-COLOR conjugated monoclonal antibody with the Coulter CD19 FITC conjugated monoclonal antibody:

procedure	mean % positive	r² value	slope	Y intercept	n
CD19 TRI-COLOR	17.1	97.4	0.94	1.36	175
CD19 FITC	16.7
CD19 TRI-COLORLinear regression	$y = 1.36 + 0.94x$

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Comparison of the CALTAG CD19 TRI-COLOR conjugated monoclonal antibody with the CALTAG CD19 R-PE conjugated monoclonal antibody:

procedure	mean % positive	r² value	slope	Y intercept	n
CD19 TRI-COLOR	17.1	97.6	0.99	-0.56	175
CD19 R-PE	16.4
CD19 TRI-COLORLinear regression	$v = -0.56 + 0.99x$

BIBLIOGRAPHY

Hsu S., Cossman J., Jaffe E.; Lymphocyte subsets in normal human lymphoid tissues. Am. J. Clin. Path. 80:21-30, 1983. 1.
Morimoto C., Levin N.L., Distaso J.A., et al: The cellular basis for the induction of antigen-specific T8-suppressor cells. 2. Eur. J. Immunol. 16:198-204, 1986.
1. Dorken B., Moller P., Pezzutto A., Schwarz-Albiez R., Moldenhauer G., B-cell antigens: CD19, in Fourth International Workshop and Conference On Human Leucocyte Differentiation Antigens, pp 34-36, Vienna, 1989.
Piruccello S.J., Johnson D.R., Reagents for flow cytomaty: Monoclopalic cell anigens, in Flow Cyonery And 4. Clinical Diagnosis, pp 56-78, pub. American Society of Clinical Pathologists, 1994.
5 Shields J.G., Righty K.P., Callard R.E., Regulation of human B cell prolification by the CD19 cell-surface glycoprotein, in Fourth International Workshop and Conference On Human Leucocyte Differentiation Antigens, pp 40-43, Vienna, 1989.
Nadler L.M., in Leukocyce Typing II, Volume 2, B cel/Leukemix Panel Workshop:Summary and Comments, Chapter 1:2-20, Springer 6. Verlag, New York, 1986.
1. Wedgwood R.I., X-linked agammaglobulinemia, in CRC Handbook Series In Clinical Laboratory Science, CRC Press, West Palm Beach Florida, pp 41-50, 1978.
Pearl E.R., Vogler L.B., Okos A.J. et al, B lymphocyce precursors in bone marrow. An analysis of normal individuals and patients with 8. antibody-deficiency states,
- J. Immunol. 120:1169-175, 1978.
Spickett G.P., Webster A.D., Farrant J., Cellular abnormalities in common variable immunodeficiency, 9.
Immunodefic. Rev. 2:199-219, 1990.
1. Small T.N., Keever C., Collins N. et al, Characterization of B cells in severe combined immunodeficiency disease, Hum. Immunol. 25:181-193, 1991.
1. Goldstein R., Laguire L.C., Douglas J. et al, Systematosus and common variable panhypogammaglobulinemia. Theoretical and practical considerations, Fed. Proc. 29:1606-1611. 1985.
1. Lokea M.L., Brosnan N.N., Back B.A., Aut K.A., Establishing opimal lymphocyce gates for immunophersonyping for flow cyometry. Cytometry 11:453-459, 1990.
Guidelines for the Performance of CD4+ T-Cell Determinations in Persons with Human Immundeticiency Virus Infection, Morbidity And 13. Mortality Weekly Report (MMWR), Volume 41/No. RR-8, May 8, 1992.
1. Nicholson J.K.A., Green T.A. and Collaborative Laboratories, Selection of anticoagulans for lymphocyce immunophenotyping. J. Immunol. Methods 165:31-35, 1993 .
ા રા Tennant J.R., Evaluation of the Trypan Blue technique for determination of cell viability, Transplanation 2:685-694, 1964.
1. Koepke J.A., Landay A.I.: Precision and Accuracy of Absolute Lymphocyte Counts,
Clin. Immunol, and Immunopath, 52:19-27, 1989.
1. Brown M.C., Hoffman R.A., Kirchanki S., Controls for flow cyometers in hematology and cethular immunology, Ann. N.Y. Acad. Sci. 468:93-103. 1986.
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Regulation Number and Section

§ 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.”