IMMULITE EPO, MODEL #'S LKEPZ & LKEP1

K983203 · Diagnostic Products Corp. · GGT · Jul 12, 1999 · Hematology

Device Facts

• Record ID: K983203
• Device Name: IMMULITE EPO, MODEL #'S LKEPZ & LKEP1
• Applicant: Diagnostic Products Corp.
• Product Code: GGT · Hematology
• Decision Date: Jul 12, 1999
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.7250
• Device Class: Class 2

Indications for Use

IMMULITE EPO is a solid-phase, two-site enzyme chemiluminescent sequential immunometric assay for use with the IMMULITE Automated Analyzer and designed for the quantitative measurement of erythropoietin (EPO) in serum or heparinized plasma. It is intended strictly for in vitro diagnostic use as an aid the diagnosis of anemias and polycythemias.

Device Story

IMMULITE EPO is an in vitro diagnostic reagent system used with the IMMULITE Automated Immunoassay Analyzer. It measures erythropoietin (EPO) levels in serum or heparinized plasma samples. The device utilizes a solid-phase, two-site sequential chemiluminescent enzyme immunometric assay. Patient samples and ligand-labeled monoclonal anti-EPO antibodies are incubated with a polystyrene bead coated with anti-ligand; subsequently, an alkaline phosphatase-labeled polyclonal anti-EPO antibody is added. A chemiluminescent substrate (phosphate ester of adamantyl dioxetane) is introduced; hydrolysis by alkaline phosphatase produces light proportional to EPO concentration. The analyzer measures photon output to quantify EPO. Used in clinical laboratories by trained personnel to assist physicians in diagnosing anemias and polycythemias by evaluating EPO levels relative to patient hematocrit and clinical condition. Results aid in differentiating primary from secondary anemias and polycythemias, supporting clinical management of conditions like chronic renal disease or polycythemia vera.

Clinical Evidence

Method comparison study performed on 130 endogenous serum samples with EPO concentrations ranging from 2.5 to 200 mU/mL. Comparison between IMMULITE EPO and the predicate radioimmunoassay (INCSTAR) yielded a linear regression of y = 0.846x + 3.818 mU/mL with a correlation coefficient (r) of 0.978. Mean values were 40.96 mU/mL for IMMULITE and 43.90 mU/mL for INCSTAR.

Technological Characteristics

Solid-phase, two-site sequential chemiluminescent enzyme immunometric assay. Solid phase consists of a polystyrene bead coated with anti-ligand. Detection uses alkaline phosphatase-labeled polyclonal anti-EPO antibody and a phosphate ester of adamantyl dioxetane chemiluminescent substrate. Operates on the IMMULITE Automated Immunoassay Analyzer. Quantitative measurement based on photon emission proportional to EPO concentration.

Indications for Use

Indicated for the quantitative measurement of erythropoietin (EPO) in serum or heparinized plasma as an aid in the diagnosis of anemias and polycythemias in patients.

Regulatory Classification

Identification

A erythropoietin assay is a device that measures the concentration of erythropoietin (an enzyme that regulates the production of red blood cells) in serum or urine. This assay provides diagnostic information for the evaluation of erythrocytosis (increased total red cell mass) and anemia.

Special Controls

*Classification.* Class II. The special control for this device is FDA's “Document for Special Controls for Erythropoietin Assay Premarket Notification (510(k)s).”

Predicate Devices

• Incstar's EPO-Trac 1251 RIA (K902639)

Submission Summary (Full Text)

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JUL 12 1999

### 510(k) Summary

This summary of safety and effectiveness information has been prepared in accordance with the requirements of SMDA 1990 and 21 CFR Part 807.92.

Diagnostic Products Corporation Name: 5700 West 96th Street Address: Los Angeles, CA 90045

Telephone Number: Facsimile Number:

Contact Person:

Date of Preparation:

Device Name Trade: Common:

Catalog Number:

Classification:

CLIA Complexity Category:

Manufacturer:

Establishment Registration Number:

Substantially Equivalent Predicate Device:

Description of the Device:

[phone] [phone]

Edward M. Levine, Ph.D. Director of Clinical Affairs

June 7, 1999

IMMULITE® EPO Reagent system for the determination of EPO in serum and heparinized plasma

LKEPZ (50 tests), LKEP1 (100 tests)

Class III device (pre-amendment) 21CFR 864.7250

Moderate, based on previous classification of analogous tests

Diagnostic Products Corporation 5700 West 96th Street Los Angeles, CA 90045-5597

DPC's Registration Number is 2017183

Incstar's EPO-Trac 1251 RIA (K902639)

IMMULITE® EPO is a solid-phase, two-site chemiluminescent sequential enzyme immunometric assay for use with the IMMULITE® Automated Immunoassay Analyzer

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Intended Use of the Device:

IMMULITE® EPO is a solid-phase, two-site enzyme chemiluminescent sequential immunometric assay for use with the IMMULITE Automated Analyzer and designed for the quantitative measurement of erythropoietin (EPO) in serum or heparinized plasma. It is intended strictly for in vitro diagnostic use as an aid the diagnosis of anemias and polycythemias.

#### Summary and Explanation of the Test:

Erythropoietin (EPO) is a glycoprotein hormone consisting of 165 amino acids, with four complex carbohydrate chains attached to the peptide at four linkage sites. It has a molecular weight of 36,000 daltons, 40% of this attributed to the carbohydrate chains. EPO is the primary regulator of erythropoiesis, stimulating the proliferation and differentiation of erythroid precursor cells in bone marrow. In mammals, the fetal liver produces nearly all of the hormone; in adults, hepatic production drops to under 10% and renal secretion accounts for over 90%. The production site is believed to be the proximal renal tubular cells or the peritubular capillary endothelial cells of the renal cortex and outer medulla. The clearance of circulating EPO has not been fully explained, but it is accomplished, in small part, by urinary excretion, and possibly also by hepatic elimination and by uptake into target cells in bone marrow.

EPO adjusts red blood cell production to meet the tissue oxygen demand. It exerts its effect in a complex feedback system, in which renal secretion of the hormone is controlled by an oxygen sensor in the kidney that responds to the partial pressure of oxygen in blood. Under conditions of increased peripheral oxygen, EPO levels diminish. This is seen after correction of hypoxia in healthy subjects (as in descent from a high elevation) and after hypertransfusion.

Anemias may be divided into two categories with respect to EPO levels in blood: those that are primary to EPO levels and those that are secondary. Primary anemias are characterized by an increase of EPO in the blood to attempt to restore red blood cell production levels to normal. Examples of anemias in which EPO levels are elevated include iron deficiency anemia, reduction of blood flow to the kidney (as in blood loss) and hemoglobinopathies with increased affinity of hemoglobin for oxygen. The EPO production rate is seen to increase exponentially with the decrease in available oxygen and with falling hematocrit in nonrenal anemias, in the latter, EPO levels 1,000 times normal have been reported.

Anemia can be secondary to inflammation, rheumatoid arthritis, neoplasm, and chronic renal disease. The "secondary anemias" may, however, be at least partly attributable to underproduction of EPO.

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A failure to produce sufficient EPO accounts for the moderate to severe anemias observed in end-stage renal disease. Decreased EPO production is attributed to destruction of renal production sites; the renal oxygen sensor may also be affected. Levels of the hormone slightly exceed the reference range at most, and are inadequate to counter the blood loss due to dialysis, shortened red blood cell life, iron and folate deficiency, impaired iron transfer to erythroid progenitor cells and other challenges faced by such patients. Anephric patients demonstrate especially low EPO levels. A few patients with chronic kidney failure, however, exhibit normal hematocrits or less serious anemia, and elevated EPO. Some of these patients have cystic kidneys or viral hepatitis; in the latter, increased EPO may have resulted from enhanced hepatic production.

Overproduction of red blood cells is called polycythemia. Polycythemias may also be divided into two categories depending on whether the condition is primary or secondary to EPO levels. In polycythemia vera, EPO levels are diminished and erythropoiesis is primary to and independent of stimulation by EPO. Variation in EPO values can be as much as tenfold for different patients with the same hematocrit.

Certain conditions may be characterized by the loss of feedback control of oxygen concentration over EPO production, causing an increase in EPO levels. These include renal cell carcinomas, in which 2% of patients demonstrate erythrocytosis, and some benign renal lesions, such as single or multiple renal cysts, renal artery stenosis and microvascular abnormalities. In addition, approximately 10% of renal transplant patients develop erythrocytosis, sometimes from autologous diseased kidney.

Secondary polycythemia is characterized by elevated EPO levels which lead to increased red blood cell mass. This condition may result from a variety of factors, including defective hemoglobin, smoking, pulmonary fibrosis, cardiac disease, tumors, and kidney stones.

When assaying EPO for the differential diagnosis of polycythemias, the possible overlap of values for secondary erythrocytosis or for polycythemia vera with those in the reference range must be considered.

### Technological Comparison to Predicate:

IMMULITE EPO is a solid-phase, two-site sequential chemiluminescent enzymelabeled immunometric assay. The solid phase, a polystyrene bead enclosed within an IMMULITE Test Unit, is coated with an anti-ligand specific for EPO.

The patient sample and a ligand-labeled monoclonal anti-EPO antibody are simultaneously introduced into the Test Unit and incubated for approximately 30 minutes at 37 ℃ with intermittent agitation. During this time, EPO in the sample binds to the anti-ligand on the solid phase.

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An alkaline phosphatase-labeled polyclonal anti-EPO antibody is introduced, and the Test Unit is incubated for another 30-minute cycle. The unbound enzyme conjugate is removed by a centrifugal wash. Substrate is then added and the Test Unit is incubated for a further 10 minutes.

The chemiluminescent substrate, a phosphate ester of adamantyl dioxetane, undergoes hydrolysis in the presence of alkaline phosphatase to yield an unstable intermediate. The continuous production of this intermediate results in the sustained emission of light, thus improving precision by providing a window for multiple readings. The bound complex and thus also the photon output, as measured by the luminometer - is proportional to the concentration of EPO in the sample.

The Incstar's EPO-Trac 1251 RIA procedure is a competitive binding, disequilibrium radioimmunoassay which utilizes recombinant human erythropoietin for both tracer and standards. Samples are incubated with the EPO-Trac primary goat antibody (goat anti-EPO) and allowed to react for 2 hours before EPO-Trac tracer labeled with iodine-125 is added Following an overnight incubation, the donkey anti-goat precipitating complex (DAG-PPT) secondary antibody is added to the specific assay test tubes (TABLE 1) The DAG-PPT is a donkey anti-goat serum that is pre-precipitated with a normal goat serum and a surfactant. The DAG-PPT is incubated with standards or samples, primary antibody and tracer, for thirty minutes before the test tubes are centrifuged to separate the bound from the unbound tracer. The unbound tracer is removed by decanting the supernatant from each test tube. The bound tracer in the remaining DAG-PPT complex pellets is counted in a gamma counter for I minute. The 122T counts are inversely proportional to the amount of EPO present in each sample.

### Method Comparison:

The IMMULITE EPO procedure was compared to a radioimmunoassay (INCSTAR) on 130 endogenous serum samples, with EPO concentrations ranging from 2.5 to 200 mU/mL. Linear regression analysis yielded the following statistics.

(IMMULITE) = 0.846 (INCSTAR) + 3.818 mU/mL r = 0.978

40.96 mU/mL (IMMULITE) Means:

43.90 mU/mL (INCSTAR)

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# Conclusion:

The data presented in this summary of safety and effectiveness is the data that the Food The Gata prescitied in this summal y or surely and equivales of the equivalence for IMMULITE® EPO.

Edward h. Lewis

Edward M. Levine, Ph.D. Director of Clinical Affairs

6/7/95

Date

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Image /page/5/Picture/0 description: The image shows the seal of the Department of Health & Human Services (HHS). The seal features a stylized caduceus-like symbol, with three abstract human figures forming a wing-like shape. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" is arranged in a circular pattern around the symbol.

DEPARTMENT OF HEALTH & HUMAN SERVICES

JUL 1.2 1999

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Edward M. Levine, Ph.D. Director of Clinical Affairs Diagnostic Products Corporation 5700 West 96th Street Los Angeles, California 90045-5597

Re: K983203

Trade Name: DPC's IMMULITE® EPO Test Kit Regulatory Class: III Product Code: GGT Dated: March 19, 1999 Received: March 22, 1999

## Dear Dr. Levine:

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 legally marketed predicate 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 Current Good Manufacturing Practice requirements, 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 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.

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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 [phone].

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 [phone]. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at [phone]. 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 [phone] or [phone], or at its internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Putman

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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510(k) Number (if known): Device Name: IMMULITE® EPO

Kq83203

Indications For Use:

IMMULITE EPO is a solid-phase, two-site sequential chemiluminescent enzyme immunometric assay for use with the IMMULITE Automated Analyzer and designed for the quantitative measurement of erythropoietin (EPO) in serum or heparinized plasma. It is intended strictly for in vitro diagnostic use as an aid in the diagnosis of anemias and polycythemias.

# (PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Division Sign-Off vision of Clinical Laboratory Devi 1983223 510(k) Numb

Prescription Use
(Per 21 CFR 801.109)

OR

Over-The-Counter Use

(Optional Format 1-2-

96)