Free Testosterone AccuBind ELISA Test System
Device Facts
| Record ID | K181017 |
|---|---|
| Device Name | Free Testosterone AccuBind ELISA Test System |
| Applicant | Monobind, Inc. |
| Product Code | CDZ · Clinical Chemistry |
| Decision Date | Jul 20, 2018 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.1680 |
| Device Class | Class 1 |
Intended Use
The Free Testosterone AccuBind® ELISA Test System is an Enzyme Immunoassay (EIA) for the quantitative measurement of free testosterone in human serum. Measurement of free testosterone is used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, impotence in males and in females; hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries and adrogenital syndromes.
Device Story
Competitive enzyme immunoassay (EIA) for quantitative measurement of free testosterone in human serum; utilizes immobilized testosterone antibody, enzyme-antigen conjugate (HRP-labeled), and native antigen from patient sample. Competitive reaction occurs between native free antigen and enzyme-antigen conjugate for limited binding sites on antibody-coated microplate. After equilibrium, unbound antigen is removed via decantation/aspiration. Substrate (TMB/hydrogen peroxide) added to produce blue color, followed by acid stop solution to yield yellow color. Microplate reader (450nm/620nm) measures absorbance; dose-response curve generated from serum calibrators determines unknown sample concentration. Used in clinical laboratories by trained personnel. Output assists clinicians in diagnosing/treating androgen-related disorders (hypogonadism, hirsutism, virilization).
Clinical Evidence
Bench testing only. Precision study (n=80 per sample) showed total CVs 7.8-12.4%. Linearity confirmed across 0.11-60 pg/mL (R^2=0.9888). LoB 0.0295 pg/mL, LoD/LoQ 0.0519 pg/mL. Interference and cross-reactivity studies showed no significant bias (>10%) for tested substances. Method comparison with predicate (n=137) yielded y=1.017x-0.244, r=0.997. Reference range established using 261 healthy subjects.
Technological Characteristics
Competitive ELISA; solid-phase microplate coated with testosterone antibody. Reagents: HRP-labeled testosterone analog conjugate, TMB substrate, stop solution. Detection: colorimetric microplate reader at 450nm. Standards: CLSI EP06-A (linearity), CLSI EP17-A (limits), CLSI EP07-A2 (interference/cross-reactivity), CLSI C28-A3 (reference ranges).
Indications for Use
Indicated for quantitative measurement of free testosterone in human serum for diagnosis and treatment of androgen-related disorders, including primary/secondary hypogonadism, impotence, hirsutism, and virilization in males and females.
Regulatory Classification
Identification
A testosterone test system is a device intended to measure testosterone (a male sex hormone) in serum, plasma, and urine. Measurement of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delayed or precocious puberty, impotence in males and, in females hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.
Predicate Devices
- EiAsy Free Testosterone EIA (k030730)
Related Devices
- K052649 — DRG SLV TESTOSTERONE ELISA TEST · Drg Intl., Inc. · Jan 27, 2006
- K014120 — TESTOSTERONE BY ENZYMEIMMUNOASSAY (EIA) · Diagnostics Biochem Canada, Inc. · Feb 8, 2002
Submission Summary (Full Text)
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Image /page/0/Picture/0 description: The image contains the logos of the Department of Health & Human Services and the Food and Drug Administration (FDA). The Department of Health & Human Services logo is on the left, and the FDA logo is on the right. The FDA logo is a blue square with the letters "FDA" in white, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue.
July 20, 2018
Monobind Inc. Anthony Shatola Quality Director 100 North Pointe Drive Lake Forest, CA 92630
Re: K181017
Trade/Device Name: Free Testosterone AccuBind ELISA Test System Regulation Number: 21 CFR 862.1680 Regulation Name: Testosterone test system Regulatory Class: Class I, reserved Product Code: CDZ Dated: June 18, 2018 Received: June 19, 2018
Dear Anthony Shatola:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and 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) that do not require approval of a premarket approval application (PMA). 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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR
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Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.
For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely.
# Kellie B. Kelm -S
for Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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## Indications for Use
510(k) Number (if known) k181017
Device Name
Free Testosterone AccuBind® ELISA Test System
Indications for Use (Describe)
The Free Testosterone AccuBind® ELISA Test System is an Enzyme Immunoassay (EIA) for the quantitative measurement of free testosterone in human serum. Measurement of free testosterone is used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, impotence in males and in females; hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries and adrogenital syndromes.
| Type of Use (Select one or both, as applicable) | |
|-----------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------|
| <div> <span> <b> ☑ </b> Prescription Use (Part 21 CFR 801 Subpart D) </span> </div> | <div> <span> ☐ Over-The-Counter Use (21 CFR 801 Subpart C) </span> </div> |
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B Monobind Inc.
The World Resource for Diagnostic Products
#### 510(k)Summarv
| 1. | Submitted by: | Monobind Inc. |
|----|----------------------------|-------------------------------------------------------|
| | Address: | 100 North Pointe Drive<br>Lake Forest, CA 92630, USA |
| | Phone: | 949-951-2665 |
| | Fax: | 949-951-3539 |
| | Website: | www.monobind.com |
| | CompanyContact: | Anthony Shatola, Quality Director |
| | Date: | July 12, 2018 |
| 2. | Product Trade Name: | Free Testosterone AccuBind® ELISA Test system |
| | FDA Submission Number: | k181017 |
| | Classification Name: | Radioimmunoassay,testosterone and dihydrotestosterone |
| | ProductCode: | CDZ |
| | Classification: | Class I, Reserved |
| | Regulation Number / Panel: | 862.1680 / Clinical Chemistry |
- Predicate Device: EiAsvFreeTestosterone EIA 3. CompanyName: Diagnostic Biochem Canada, Inc. 510(k)Number: k030730
- A. TestPrinciple:
Monobind Inc. Free Testosterone test system uses a competitive enzyme immunoassay. The essential reagents required for a solid phase enzyme immunoassay include immobilized antibody, enzyme-antigen conjugate and native antigen. Upon mixing immobilized antibody, enzyme-antigen conjugate and a serum containing the native antigen, a competition reaction results between the native antigen and the enzyme-antigen conjugate for a limited number of insolubilized binding sites. After equilibrium is attained, the antibody-bound fraction is separated from unbound antigen by decantation or aspiration. The enzyme activity in the antibody-bound fraction is inversely proportional to the native antigen concentration. By utilizing several different serum references of known antigen concentration, a dose response curve can be generated from which the antigen concentration of an be ascertained. The serum calibrators are prepared in human serum matrix. The enzyme-antigen conjugate is labelled with horseradish peroxidase (HRP) and the substrate reagent contains tetramethylbenzidine (TMB), a blue color is produced. The reaction is stopped with addition of an acid and a yellow color is developed. The plate is read in a microtiter plate reader at 450nm.
- B. KitDescription:
The kit consists of seven (7) vials of serum reference calibrators for Free Testosterone with two (2) controls (one low and one high); one (1) vial of Testosterone (Analog)-horseradish peroxidase (HRP) conjugate in a protein stabilizing matrix; one 96-well testosterone antibody-coated microplate; one (1) vial of concentrated wash solution; two (2) vials for tetramethy(benzidine (TMB) substrate solution preparation; and one (1) vial of stop reaction solution.
- C. Indications for Use:
The Free Testosterone AccuBind® ELISA Test System is an Enzyme Immunoassay (EIA) for the quantitative measurement of free testosterone in human serum. Measurement of free testosterone is used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, impotence in males and in females; hirsutism (excessive hair) and virilization (masculinization)due to tumors, polycystic ovaries and adrogenital syndromes.
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Comparison with Predicate (EiAsy Free Testosterone EIA):
Similarities and Differences
| | Predicate | Monobind |
|-------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | The direct quantitative<br>determination of Free<br>Testosterone byenzyme<br>immunoassay in human serum | Same |
| Antibody | Utilizes a highly specific rabbit<br>polyclonal antibody at a low<br>binding capacity | Same |
| Detection Method | Microplate colorimetricreader | Same |
| Test Principle | Competitive Enzyme<br>Immunoassay | Same |
| Sample Type | Human Serum | Same |
| Key Reagents | | |
| Calibrators | Six (6) vials containing<br>testosterone in human serum<br>with preservative | Seven (7) vials containing<br>testosterone in human serum with<br>preservative |
| Control | Two (2) vials containing<br>testosterone in human serum | Three (3) vials containing<br>testosterone in human serum |
| Calibrator/Control Matrix | Human Serum | Same |
| HRP conjugate | Testosterone-HRP conjugate<br>in buffer | Same |
| Wash Buffer Concentrate | Concentrated (10x) buffer with<br>preservatives | Same |
| TMB Substrate | One vial containing<br>tetramethylbenzidine and<br>hydrogen peroxide in buffer | Two (2) vials: one containing<br>tetramethylbenzidine, one<br>containing hydrogenperoxide |
| Microplate | Antibody coated plate | Same |
| Specimen treatment | Direct system; no specimen<br>pretreatmentnecessary | Same |
| Measuring range | 0.018-60pg/ml | 0.11-60pg/ml |
| Expected Range of values<br>(pg/ml) | Males / 20-39: 9.1-32.2<br>Males / 40-59: 5.7-30.7<br>Males / ≥60: 5.9-27.0<br>Females / 20-39: 0.1-6.3<br>Females / 40-59: 0.3-5.0<br>Females / ≥60: 0.5-3.9 | Males / 20-39: 9.2-34.6<br>Males / 40-59: 6.1-30.3<br>Males / ≥60: 6.1-27.9<br>Females / 20-39: 0.2-6.1<br>Females / 40-59: 0.3-4.4<br>Females / ≥60: 0.5-3.4 |
TestSummary
- 1. AnalyticalPerformance
- Precision a.
This study was conducted during 20 days of testing. The human serum and control samples were tested in duplicate, two times per day for a total of 80 measurements per sample. Three (3) different reagent lots, three (3) serum pools, and three (3) controls were used for the study (low, medium, and high concentration). The results of a representative lot are shown below:
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| Lot 1<br>N=32 | Mean (pg/ml) | Within-Run | | Total | |
|---------------|--------------|------------|------|-------|-------|
| | | SD | CV | SD | CV |
| Control 1 | 2.51 | 0.09 | 3.7% | 0.20 | 7.8% |
| Control 2 | 10.98 | 0.40 | 3.6% | 0.96 | 8.7% |
| Control 3 | 22.72 | 0.83 | 3.6% | 2.18 | 9.6% |
| Serum 1 | 0.98 | 0.06 | 5.9% | 0.12 | 12.4% |
| Serum 2 | 4.53 | 0.26 | 5.7% | 0.36 | 8.0% |
| Serum 3 | 53.62 | 4.24 | 7.9% | 4.32 | 8.1% |
The results of the combined lot precision are shown below:
| | Mean<br>Value<br>(pg/ml) | Within-Run Precision | | Within-Kit Precision | | Total Precision (n=80) | |
|-----------|--------------------------|----------------------|------|----------------------|------|------------------------|------|
| | | SD | CV% | SD | CV% | SD | CV% |
| Control 1 | 2.48 | 0.11 | 4.57 | 0.20 | 8.20 | 0.21 | 8.51 |
| Control 2 | 11.04 | 0.47 | 4.23 | 0.84 | 2.60 | 0.87 | 8.00 |
| Control 3 | 23.24 | 1.00 | 4.31 | 1.80 | 7.73 | 1.83 | 7.95 |
| Patient 1 | 0.97 | 0.05 | 4.88 | 0.09 | 9.14 | 0.08 | 9.43 |
| Patient 2 | 4.62 | 0.23 | 4.88 | 0.32 | 6.89 | 0.33 | 7.20 |
| Patient 3 | 54.66 | 3.25 | 5.95 | 3.92 | 7.17 | 4.13 | 7.55 |
#### Linearity b.
Sample preparations of 11 concentrations were prepared (0.10-64.91 pg/mL) and assayed per CLSI EP06-A guidance, Evaluation of the Linearity of Quantitative Measurement Procedures. The linear regression obtained is as follows: y=1.0149x-0.6028, R =0.9888
- c. Recovery
A study was performed to evaluate the recovery of the test system. Five serum samples containing different levels of endogenous testosterone were spiked with known concentrations of testosterone throughout the measuring range. The samples were measured in replicates of three and the average concentration was compared against the expected value. The recovery results are summarized below:
| Sample | Endogenous Concentration<br>(pg/ml) | Average % Recovery |
|-------------------|-------------------------------------|--------------------|
| Patient Sample #1 | 0.57 | 100.2% |
| Patient Sample #2 | 0.24 | 102.4% |
| Patient Sample #3 | 5.65 | 98.7% |
| Patient Sample #4 | 0.91 | 104.0% |
| Patient Sample #5 | 0.05 | 105.2% |
Image /page/5/Picture/9 description: The image shows the logo for Monobind Inc. The logo also includes the text "ISO 13485 & ISO 9001 Certified Company". There are also four small icons above the text. The icons are a magnifying glass, a microscope, a box, and an organizational chart.
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- d. ReagentStability/SampleStability
Real time stability studies are conducted to determine the reagent and kit shelf life (expiration date). Expiration date of the Monobind Free Testosterone AccuBind® ELISA Test System is determined by results of shelf studies and is based on the reagent that has the shorter assigned expirationdate.
- 1. The long term stability for kit reagents stored at 2-8 ℃ is 2 years from the manufacturingdate.
- Open vial, Free Testosterone Calibrator and controls, Free Testosterone Enzyme 2. Reagentstabilitydetermination
| Condition | Stability | StorageTemperature |
|-----------------------------|-----------|--------------------|
| Open Kit Stability | 61 days | 2-8 °C |
| FreeTestosteroneCalibrators | 61 days | 2-8 °C |
| FreeTestosteroneControls | 61 days | 2-8 °C |
| Free Testo Enzyme Reagent | 61 days | 2-8 °C |
| TMB Substrate(A&B) | 61 days | 2-8 °C |
| Antibody Coated Plate | 61 days | 2-8 °C |
SampleStability
| Condition | Stability | StorageTemperature |
|---------------------|-----------|--------------------|
| Serum Sample | 61 days | 2-8°C |
| Frozen Serum Sample | 31 days | -20°C |
- Expected Reference Values e.
The reference range was established according to literature and verified utilizing 261 male and female serum samples. The reference range was calculated using CLSI C28-A3 as a guide. The following table indicates the summary of the results.
| Population | N | Reference Range (pg/ml) |
|----------------|----|-------------------------|
| Male / 20-39 | 45 | 9.2-34.6 |
| Male / 40-59 | 43 | 6.1-30.3 |
| Male / ≥60 | 43 | 6.1-27.9 |
| Female/20-39 | 44 | 0.2-6.1 |
| Female / 40-59 | 42 | 0.3-4.4 |
| Female / ≥60 | 44 | 0.5-3.4 |
- f. DetectionLimits
The LOB (limit of the blank), the LOD (limit of detection) and the LOQ (limit of quantitation) were determined in accordance with CLSI EP 17-A guideline, Protocols for Determination of Limits of Detection).
| Limit of the Blank (LoB) | Limit of Detection (LoD) | Limit of Quantitation (LoQ) |
|--------------------------|--------------------------|-----------------------------|
| 0.0295pg/ml | 0.0519pg/ml | 0.0519pg/ml |
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## g. CrossReactivity
Cross reactivity was determined by testing those compounds most likely to interfere with the Monobind Free Testosterone ELISA Test System. The specificity of the assay was determined in accordance with CLSI EP07-A2. Cross-reactivity was determined using the following equation: observed value - unspiked value/ concentration of cross-reactant x100%. Significant cross reactivity is defined as > 10% difference. The results of the cross-reactivity study are as follows.
| Sample | Conc.<br>(ng/ml) | Male Serum<br>Spiked Samples | | Blank Serum<br>Spiked Samples | |
|-------------------------------|------------------|------------------------------|---------------------|-------------------------------|---------------------|
| | | Conc<br>(pg/ml) | Cross<br>Reactivity | Conc.<br>(pg/ml) | Cross<br>Reactivity |
| Base Pool | 0 | 7.408 | - | 0 | - |
| 11-Deoxycortisol | 1000 | 7.269 | 0.000% | 0.000 | ND |
| 11-KetoTestosterone | 10 | 72.139 | 0.647% | 51.855 | 0.519% |
| 11β-Hydroxytestosterone | 100 | 72.815 | 0.065% | 54.000 | 0.054% |
| 17a-ethynyl estradiol | 1000 | 7.351 | 0.000% | 0.000 | ND |
| 17α-Estradiol | 1000 | 7.282 | 0.000% | 0.153 | 0.000% |
| 17β-Estradiol | 100 | 7.363 | 0.000% | 0.000 | ND |
| 17-Hydroxypregnenolone | 1000 | 7.35 | 0.000% | 0.000 | ND |
| 17-Hydroxprogesterone | 10 | 7.969 | 0.000% | 1.012 | 0.000% |
| 3-EstriolGluc | 1000 | 7.825 | 0.000% | 0.000 | ND |
| 3-EstriolSul | 1000 | 11.729 | 0.000% | 0.000 | ND |
| 3β-Androstanediol | 500 | 7.917 | 0.000% | 0.000 | ND |
| 5a-Dihydrotestosterone | 100 | 60.975 | 0.054% | 42.382 | 0.042% |
| Aldosterone | 8000 | 12.80 | 0.000% | 4.919 | 0.000% |
| AmitriptylHCI | 1000 | 7.301 | 0.000% | 0.000 | ND |
| Androsterone | 1000 | 8.061 | 0.000% | 0.000 | ND |
| Andronstenedione | 1000 | 50.552 | 0.004% | 19.248 | 0.002% |
| Clomiphene Citrate | 1000 | 7.263 | 0.000% | 0.000 | ND |
| Corticosterone | 1000 | 10.099 | 0.000% | 0.777 | 0.000% |
| Cortisone | 1000 | 9.90 | 0.000% | 0.478 | 0.000% |
| Cortisol | 1000 | 7.316 | 0.000% | 0.298 | 0.000% |
| Cyproteroneacetate | 1000 | 10.70 | 0.000% | 0.000 | ND |
| D-5-Androstene-3β,17β-diol | 1000 | 7.782 | 0.000% | 0.000 | ND |
| Danazol | 1000 | 12.30 | 0.000% | 0.000 | ND |
| DHEA | 100000 | 7.311 | 0.000% | 7.457 | 0.000% |
| DHEA-S | 1000 | 7.45 | 0.000% | 0.238 | 0.000% |
| Desogestrel | 100 | 7.436 | 0.000% | 0.000 | ND |
| Dexamethasone | 1000 | 7.381 | 0.000% | 0.000 | ND |
| Epistestosterone | 1000 | 21.612 | 0.001% | 8.215 | 0.001% |
| Estriol | 1000 | 7.368 | 0.000% | 0.122 | 0.000% |
| Estrone | 1000 | 7.679 | 0.000% | 0.036 | 0.000% |
| Ethisterone | 1000 | 8.597 | 0.000% | 0.003 | 0.000% |
| Sample | Conc.<br>(ng/ml) | Conc<br>(pg/ml) | Cross<br>Reactivity | Conc.<br>(pg/ml) | Cross<br>Reactivity |
| Ethynodiol | 1000 | 8.042 | 0.000% | 0.114 | 0.000% |
| Ethynodioldiacetate | 50 | 7.563 | 0.000% | 0.000 | ND |
| Flunisolide | 1000 | 7.456 | 0.000% | 0.000 | ND |
| Fluoxymesterone | 1000 | 7.450 | 0.000% | 0.000 | ND |
| Lynestrol | 1000 | 7.395 | 0.000% | 0.000 | ND |
| Medoxyprogesterone<br>acetate | 1000 | 7.426 | 0.000% | 0.000 | ND |
| Methyl Testosterone | 100 | 7.163 | 0.000% | 0.000 | ND |
| Mestranol | 1000 | 7.338 | 0.000% | 0.000 | ND |
| Norethindrone | 50 | 7.541 | 0.000% | 0.000 | ND |
| Norethinodroneacetate | 50 | 7.428 | 0.000% | 0.000 | ND |
| Norgestimate | 1000 | 7.478 | 0.000% | 0.000 | ND |
| Norgestrel (Levonorgestrel) | 50 | 7.463 | 0.000% | 0.000 | ND |
| Norethynodrel | 50 | 7.544 | 0.000% | 0.000 | ND |
| Oxymetholone | 100 | 7.401 | 0.000% | 0.000 | ND |
| Prednisolone | 1000 | 7.828 | 0.000% | 0.000 | ND |
| Prednisone | 800 | 10.442 | 0.000% | 0.966 | 0.000% |
| Progesterone | 1000 | 11.025 | 0.000% | 0.248 | 0.000% |
| Salbutamol | 1000 | 8.026 | 0.000% | 0.000 | ND |
| Spironolactone | 1000 | 7.310 | 0.000% | 0.112 | 0.000% |
| Stanozolol | 1000 | 7.442 | 0.000% | 0.081 | 0.000% |
| Testosteroneenanthate | 100 | 7.304 | 0.000% | 0.044 | 0.000% |
| Testosterone SO4 | 1000 | 75.970 | 0.004% | 28.182 | 0.003% |
| Testosterone Propionate | 1000 | 9.589 | 0.000% | 1.008 | 0.000% |
| Triamcinolone | 50 | 7.544 | 0.000% | 0.043 | 0.000% |
Image /page/7/Picture/5 description: The image shows the Monobind Inc. logo, which includes a magnifying glass, a microscope, a box, and a flowchart. The text "Monobind Inc." is located below the microscope. The text "ISO 13485 & ISO 9001 Certified Company" is located below the logo. The image is in a light blue color.
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An additional study was performed to evaluate the cross-reactivity effects of testosterone cypionate and testosterone undecanoate. Aliquots from pool of human serum with a free testosterone concentration of 38.4 pg/mL were spiked with 12 ng/mL of testosterone cypionate and testosterone undecanoate. Cross-reactivity was determined using the following equation: observed value - unspiked value/ concentration of cross-reactant x100%. Significant crossreactivity is defined as > 10% difference. The results are summarized in the chart below:
| Sample | Conc.<br>(ng/ml) | Conc.<br>(pg/ml) | Cross<br>Reactivity | Conc.<br>(pg/ml) | Cross<br>Reactivity |
|--------------------------|------------------|------------------|---------------------|------------------|---------------------|
| Testosterone Cypionate | 12 | 38.685 | 0.002% | 0.021 | 0.000% |
| Testosterone Undecanoate | 12 | 38.262 | -0.001% | 0.015 | -0.001% |
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#### Method Comparison Studies 3.
Tests were conducted for comparison between the Free Testosterone ELISA Test System and the predicate assay, EiAsy Free Testosterone EIA. The free testosterone concentrations of 137 samples ranging from 0.11-59.63 pg/ml were compared. Comparison of the Monobind Free Testosterone AccuBind® ELISA Test System (new device) and the DBC EiAsy Free Testosterone EIA (predicate) show the following results:
| Method | Least Square Regression<br>Analysis | CorrelationCoefficient |
|-----------------|-------------------------------------|------------------------|
| This Method (y) | $y= 1.017x - 0.244$ | 0.997 |
| Reference(x) | | |
#### Interferences 4.
Using CLSI-A2 Interference Testing in Clinical Chemistry as a guide, potential interferents were tested utilizing charcoal-stripped human serum spiked with known concentrations of interferent.
The following results of % binding values even at higher than normal interferent levels indicate that there is no significant binding of the free testosterone-HRP conjugate.
| Substance | Highest concentration at which no significant<br>interference was observed |
|-----------------------|----------------------------------------------------------------------------|
| Acetaminophen | 20 mg/dl |
| Acetylcysteine | 150mg/dl |
| Ascorbic Acid | 6 mg/dl |
| Bilirubin Conjugated | 15 mg/dl |
| BilirubinUnconjugated | 20 mg/dl |
| Biotin | 100 ng/ml |
| Caffeine | 6 mg/dl |
| Cholesterol | 503mg/dl |
| Creatine | 30 mg/dl |
| Dextran | 5000 mg/dl |
| Digoxin | 6.1 ng/ml |
| Doxycycline | 50 mg/L |
| Erythromycin | 6 mg/dl |
| Gentamicin | 1 mg/dl |
| HAMA | 440 ng/ml |
| Heparin | 3 U/ml |
| Hemoglobin | 500mg/dl |
| Human Serum Albumin | 2.5 g/dl |
| Ibuprofen | 50 mg/dl |
| Immunoglobulin G | 4 g/dl |
| Levodopa | 20 mg/L |
| Lidocaine | 1.2mg/dl |
| Lipemia(glycerides) | 1000 mg/dl |
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| Substance | Highest concentration at which no significant<br>interference was observed |
|-------------------|----------------------------------------------------------------------------|
| Methyldopa | 20 mg/L |
| Nicotine | 0.1 mg/dl |
| Phenobarbital | 15 mg/dl |
| Protein:Total | 10.5 g/dl |
| Rheumatoid Factor | 1110 IU/ml |
| Salicylic Acid | 60 mg/dl |
| SHBG | 200 µg/ml |
| Triglycerides | 900 mg/dl |
| Urea | 500mg/dl |
ConcludingStatement:
Taken together, the performance characteristics, comparison studies with a predicate device and acceptable statistical performance studies in this 510(k) Class I, reserved submission demonstrates that the Monobind Free Testosterone AccuBind® ELISA Test System is safe and effective for its intended use and is substantially equivalent to the predicate device.
Image /page/10/Picture/3 description: The image shows the Monobind Inc. logo, which includes a magnifying glass, a microscope, a box, and an organizational chart. Below the logo is the text "Monobind Inc." followed by "ISO 13485 & ISO 9001 Certified Company". The text indicates that Monobind Inc. is certified under ISO 13485 and ISO 9001 standards.
