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 androgenital syndromes.

Device Description

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.

AI/ML Overview

The provided document describes the analytical performance of the Monobind Inc. Free Testosterone AccuBind® ELISA Test System, an in vitro diagnostic device, rather than an AI/ML-driven device. Therefore, many of the requested criteria often associated with AI/ML device studies (e.g., number of experts for ground truth, MRMC studies, human-in-the-loop performance, training set details) are not applicable to this type of medical device.

However, I can extract the relevant information regarding the device's acceptance criteria and the study that proves it meets them as presented in the document.

Here's the breakdown:

1. Table of Acceptance Criteria and Reported Device Performance

For this type of in-vitro diagnostic device, "acceptance criteria" are typically defined by regulatory standards and good laboratory practices (e.g., CLSI guidelines). The document presents analytical performance data against these established industry benchmarks rather than explicit numerical acceptance criteria beyond what is internally defined for successful assay development and validation (e.g., precision specifications).

Performance Metric	Acceptance Criteria (Implicit from Industry Standards/Good Practice)	Reported Device Performance (Representative Lot / Combined Lot)
Precision	Low %CV for within-run and total precision across concentrations.	Within-Run CV: 3.6-7.9% (Representative Lot), 4.23-5.95% (Combined Lot)
		Total CV: 7.8-12.4% (Representative Lot), 7.20-9.43% (Combined Lot)
Linearity (Accuracy)	High correlation coefficient (R) and slope close to 1.	y = 1.0149x - 0.6028, R = 0.9888 (Excellent linearity)
Recovery	Percent recovery close to 100% for spiked samples.	98.7% - 105.2% (Across 5 patient samples)
Reagent/Kit Stability	Demonstrates stability for stated shelf life and open-vial stability.	2 years (unopened kit), 61 days (open kit, calibrators, controls, enzyme reagent, TMB, plate)
Sample Stability	Demonstrates stability for specified storage conditions.	61 days (serum at 2-8°C), 31 days (frozen serum at -20°C)
Detection Limits	Defined LOB, LOD, LOQ based on CLSI EP17-A.	LoB: 0.0295 pg/ml; LoD: 0.0519 pg/ml; LoQ: 0.0519 pg/ml
Cross-Reactivity	Generally low cross-reactivity (<10% difference) with specified interfering substances.	Generally 0.000% for most compounds; highest reported up to 0.647% (for 11-KetoTestosterone). No significant cross-reactivity (>10% difference) observed.
Method Comparison	Strong correlation with predicate device (high R, slope near 1, low intercept).	y = 1.017x - 0.244, Correlation Coefficient = 0.997 (Excellent agreement)
Interferences	No significant interference at specified concentrations for various substances.	Most substances showed no significant interference at the tested concentrations.

2. Sample Size Used for the Test Set and Data Provenance

Precision Study:
- Sample Size: For the representative lot, 3 control samples were tested, with 80 measurements per sample (duplicate, two times per day for 20 days). 3 serum pools were also tested in the same manner (N=32 readings for the representative lot table, N=80 for combined lot table). For the combined lot precision, 3 controls and 3 patient samples (serum pools) were used, with a total of 80 measurements per sample.
- Data Provenance: Not explicitly stated, but typical for in vitro diagnostic device validation, these would be controlled laboratory studies using clinical samples (serum pools and control materials). No mention of country of origin or retrospective/prospective clinical data for the performance evaluation (this differs from the reference range determination).
Linearity Study: 11 concentrations of sample preparations.
Recovery Study: 5 serum samples (containing different levels of endogenous testosterone).
Reference Range Determination: 261 male and female serum samples.
- Data Provenance: Not specified for the 261 samples, but typically these samples are collected under ethical guidelines from a relevant population.
Cross-Reactivity Study: Specific compounds were tested with male serum spiked samples and blank serum spiked samples. No specific number of replicates per compound is given, but "aliquots from pool of human serum" are mentioned for testosterone cypionate and undecanoate.
Method Comparison Study: 137 samples.
- Data Provenance: Not explicitly stated, but these would be clinical samples with varying testosterone levels.
Interferences Study: Charcoal-stripped human serum spiked with known concentrations of interferent.

3. Number of Experts Used to Establish the Ground Truth and Qualifications of Experts

Not Applicable. For an in-vitro diagnostic assay for quantitative measurement of an analyte like Free Testosterone, the "ground truth" is established by the analytical method itself (the assay's ability to accurately measure the target analyte) and validated against reference methods or calibrated materials. There are no human "experts" establishing image-based ground truth as would be the case for AI/ML diagnostic tools. The predicate device serves as a comparative "ground truth" for method comparison.

4. Adjudication Method for the Test Set

Not Applicable. This is an in-vitro diagnostic assay, not an AI/ML system requiring human adjudication of results. The results are quantitative measurements.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No. This is an in-vitro diagnostic assay, not an imaging AI/ML device that assists human readers. Therefore, an MRMC study is not relevant or performed.

6. Standalone (Algorithm Only) Performance

Yes (inherently). The device itself is the "standalone" measurement system. Its performance (precision, linearity, recovery, sensitivity, specificity, interference) is evaluated on its own. There isn't a separate "human-in-the-loop" component as would be found in an AI-assisted diagnostic workflow.

7. The Type of Ground Truth Used

The ground truth for this device's performance validation is established through:
* Known Concentrations: For precision (control materials with known values), linearity (prepared concentrations), recovery (spiked samples with known additions), detection limits (analyzing blanks and low-concentration samples).
* Reference Methods/Predicate Device: For method comparison, the predicate device (EiAsy Free Testosterone EIA) serves as the comparator.
* Established Analytical Principles: The fundamental biochemical reactions and measurement principles of the ELISA platform.
* CLSI Guidelines: Adherence to established Clinical and Laboratory Standards Institute (CLSI) guidelines (e.g., EP06-A, C28-A3, EP17-A, EP07-A2) for validation studies provides the framework for defining acceptable performance.

8. The Sample Size for the Training Set

Not Applicable. This is an in-vitro diagnostic assay, not an AI/ML device that requires a distinct "training set" in the context of machine learning model development. The development and optimization of the assay's reagents and protocol are analogous to "training" in a general sense, but no specific numerical sample size is defined as a training set for an algorithm.

9. How the Ground Truth for the Training Set Was Established

Not Applicable. As above, there is no AI/ML "training set." The "ground truth" for developing such an assay comes from fundamental biochemical understanding, chemical synthesis of reagents, and iterative optimization of assay conditions to achieve desired analytical performance characteristics. This involves standard laboratory development practices common to IVD manufacturing.

Summary

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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)

Type of Use (Select one or both, as applicable)
☑ Prescription Use (Part 21 CFR 801 Subpart D)	☐ Over-The-Counter Use (21 CFR 801 Subpart C)

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B Monobind Inc.

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510(k)Summarv

1.	Submitted by:	Monobind Inc.
	Address:	100 North Pointe DriveLake 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 quantitativedetermination of FreeTestosterone byenzymeimmunoassay in human serum	Same
Antibody	Utilizes a highly specific rabbitpolyclonal antibody at a lowbinding capacity	Same
Detection Method	Microplate colorimetricreader	Same
Test Principle	Competitive EnzymeImmunoassay	Same
Sample Type	Human Serum	Same
Key Reagents
Calibrators	Six (6) vials containingtestosterone in human serumwith preservative	Seven (7) vials containingtestosterone in human serum withpreservative
Control	Two (2) vials containingtestosterone in human serum	Three (3) vials containingtestosterone in human serum
Calibrator/Control Matrix	Human Serum	Same
HRP conjugate	Testosterone-HRP conjugatein buffer	Same
Wash Buffer Concentrate	Concentrated (10x) buffer withpreservatives	Same
TMB Substrate	One vial containingtetramethylbenzidine andhydrogen peroxide in buffer	Two (2) vials: one containingtetramethylbenzidine, onecontaining hydrogenperoxide
Microplate	Antibody coated plate	Same
Specimen treatment	Direct system; no specimenpretreatmentnecessary	Same
Measuring range	0.018-60pg/ml	0.11-60pg/ml
Expected Range of values(pg/ml)	Males / 20-39: 9.1-32.2Males / 40-59: 5.7-30.7Males / ≥60: 5.9-27.0Females / 20-39: 0.1-6.3Females / 40-59: 0.3-5.0Females / ≥60: 0.5-3.9	Males / 20-39: 9.2-34.6Males / 40-59: 6.1-30.3Males / ≥60: 6.1-27.9Females / 20-39: 0.2-6.1Females / 40-59: 0.3-4.4Females / ≥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 1N=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:

	MeanValue(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(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.(ng/ml)	Male SerumSpiked Samples		Blank SerumSpiked Samples
		Conc(pg/ml)	CrossReactivity	Conc.(pg/ml)	CrossReactivity
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.(ng/ml)	Conc(pg/ml)	CrossReactivity	Conc.(pg/ml)	CrossReactivity
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
Medoxyprogesteroneacetate	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.(ng/ml)	Conc.(pg/ml)	CrossReactivity	Conc.(pg/ml)	CrossReactivity
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 RegressionAnalysis	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 significantinterference 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

{10}------------------------------------------------

Substance	Highest concentration at which no significantinterference 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.

Regulation Number and Section

§ 862.1680 Testosterone test system.

(a)
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.(b)
Classification. Class I.