ST AIA-PACK SHBG, ST AIA-PACK SHBG Calibrator Set

K143075 · Tosoh Bioscience, Inc. · CDZ · Jul 2, 2015 · Clinical Chemistry

Device Facts

• Record ID: K143075
• Device Name: ST AIA-PACK SHBG, ST AIA-PACK SHBG Calibrator Set
• Applicant: Tosoh Bioscience, Inc.
• Product Code: CDZ · Clinical Chemistry
• Decision Date: Jul 2, 2015
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1680
• Device Class: Class 1

Intended Use

ST AIA-PACK SHBG is designed for In Vitro Diagnostic Use Only for the quantitative measurement of sex hormone binding globulin (SHBG) in human serum or Na heparinized plasma on Tosoh AIA System Analyzers. The ST AIA-PACK SHBG assay is intended for use as an aid in the diagnosis of androgen disorders. The ST AIA-PACK SHBG Calibrator Set is intended for In Vitro Diagnostic Use Only for the calibration of the ST AIA-PACK SHBG assay.

Device Story

The ST AIA-PACK SHBG is a two-site immunoenzymometric assay performed in unit-dose test cups on Tosoh AIA System Analyzers. The device uses human serum or Na heparinized plasma as input. Inside the test cup, SHBG in the sample binds to monoclonal antibody immobilized on magnetic beads and enzyme-labeled monoclonal antibody. After washing to remove unbound components, a fluorogenic substrate (4-methylumbelliferyl phosphate) is added. The resulting fluorescence is directly proportional to the SHBG concentration. A standard curve is constructed using the ST AIA-PACK SHBG Calibrator Set to calculate unknown sample concentrations. The device is used in clinical laboratory settings by trained personnel. The output provides quantitative SHBG levels, which clinicians use as an aid in diagnosing androgen disorders, potentially guiding patient management and treatment decisions.

Clinical Evidence

No clinical diagnostic studies performed. Evidence consists of analytical performance data: precision (total CV 2.3-6.8%), linearity (R^2=0.9924-0.9997), LoQ (0.20 nmol/L), and method comparison against predicate (n=126, slope 0.949, R=0.991). Reference ranges established for pre/post-menopausal women and males (21-49 and >=50 years) using 122-123 subjects per group.

Technological Characteristics

Two-site immunoenzymometric assay. Components: unit dose test cups with lyophilized magnetic beads coated with anti-SHBG mouse monoclonal antibody and enzyme-labeled anti-SHBG mouse monoclonal antibody. Energy source: Tosoh AIA System Analyzer (fluorescence detection). Connectivity: System-integrated. Calibration: 6-point calibrator set traceable to NIBSC 08/266. Software: Embedded firmware for instrument control and data processing.

Indications for Use

Indicated for the quantitative measurement of sex hormone binding globulin (SHBG) in human serum or Na heparinized plasma to aid in the diagnosis of androgen disorders.

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

• Architect SHBG Reagent Kit (k060818)
• Architect SHBG Calibrator Kit (k060818)

Submission Summary (Full Text)

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract symbol that resembles a stylized caduceus, with three figures or lines intertwined.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

July 2, 2015

TOSOH BIOSCIENCE, INC. ROBERT WICK REGULATORY SPECIALIST 6000 SHORELINE COURT SUITE 101 SOUTH SAN FRANCISCO CA 94080

Re: K143075

Trade/Device Name: ST AIA-PACK SHBG, ST AIA-PACK SHBG Calibrator Set Regulation Number: 21 CFR 862.1680 Regulation Name: Testosterone Test System Regulatory Class: I. Reserved Product Code: CDZ, JIT Dated: June 22, 2015 Received: June 23, 2015

Dear Robert Wick:

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 Parts 801 and 809); medical device reporting (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 Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. 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 the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

# Katherine Serrano -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) K143075

Device Name ST AIA-PACK SHBG, ST AIA-PACK SHBG Calibrator Set

Indications for Use (Describe)

ST AIA-PACK SHBG is designed for In Vitro Diagnostic Use Only for the quantitative measurement of sex hormone binding globulin (SHBG) in human serum or Na heparinized plasma on Tosoh AIA System Analyzers. The ST AIA-PACK SHBG assay is intended for use as an aid in the diagnosis of androgen disorders.

The ST AIA-PACK SHBG Calibrator Set is intended for In Vitro Diagnostic Use Only for the calibration of the ST AIA-PACK SHBG assay.

| Type of Use (Select one or both, as applicable) | |
|-------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------|
| <span style="font-size: 16px;"> <input checked="true" type="checkbox"/> Prescription Use (Part 21 CFR 801 Subpart D) </span> | <span style="font-size: 16px;"> <input type="checkbox"/> Over-The-Counter Use (21 CFR 801 Subpart C) </span> |

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# 510(k) Summary

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# 510(k) Summary

# K143075

# ST AIA-PACK SHBG

| Date: | June 30, 2015 |
|-------------------|----------------------------------------------------------------------------------------------------------|
| Submitter: | Tosoh Bioscience, Inc<br>3600 Gantz Road<br>Grove City, OH 43123 |
| Contact Person: | Robert L. Wick<br>Regulatory Specialist<br>6000 Shoreline Ct., Ste. 101<br>South San Francisco, CA 94080 |
| | Phone: [phone] |
| | Fax: [phone] |
| | Email: [e-mail] |
| Device Name: | ST AIA-PACK SHBG |
| Classification: | Class I, reserved<br>CDZ<br>Clinical Chemistry<br>21 CFR 862.1680 |
| Device Name: | ST AIA-PACK SHBG Calibrator Set |
| Classification | Class II<br>JIT<br>Clinical Chemistry<br>21 CFR 862.1150 |
| Predicate Device: | k060818<br>Abbott/ BIOKIT S.A.<br>ARCHITECT SHBG REAGENT KIT, ARCHITECT<br>SHBG CALIBRATOR KIT |

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# 510(k) Summary

# ST AIA-PACK SHBG

According to the requirements of 21 CFR 807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.

#### Device Description:

The ST AIA-PACK SHBG is a two-site immunoenzymometric assay which is performed entirely in the ST AIA-PACK SHBG test cups. SHBG present in the test sample is bound with monoclonal antibody immobilized on a maqnetic solid phase and enzyme-labeled monoclonal antibody in the test cups. The magnetic beads are washed to remove unbound enzyme-labeled antibody and are then incubated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is directly proportional to the SHBG concentration in the test sample. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.

The following materials are required to perform SHBG analysis using the ST AIA-PACK SHBG (Cat. No. 025238) on the Tosoh AIA System Analyzer.

| ST AIA-PACK SHBG Calibrator Set | 025338 |
|-------------------------------------------|--------|
| ST AIA-PACK SHBG Sample Diluting Solution | 025538 |

#### Device Intended Use:

ST AIA-PACK SHBG is designed for In Vitro Diagnostic Use Only for the quantitative measurement of sex hormone binding globulin (SHBG) in human serum or Na heparinized plasma on Tosoh AIA System Analyzers. The ST AIA-PACK SHBG assay is intended for use as an aid in the diagnosis of androgen disorders.

Calibrators:

The ST AIA-PACK SHBG Calibrator Set is intended for In Vitro Diagnostic Use Only for the calibration of the ST AIA-PACK SHBG assay.

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# Substantial Equivalence:

# Comparison between the Tosoh ST AIA-PACK SHBG and the Abbott Architect SHBG Immunoassay (k060818)

## Similarities

| Parameter | ST AIA-PACK SHBG | Abbott Architect SHBG Kit<br>(k060818) |
|------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended use | For the quantitative<br>measurement of SHBG in<br>human serum and plasma. | For the quantitative<br>measurement of SHBG in<br>human serum and plasma. |
| Indications for Use | ST AIA-PACK SHBG is<br>designed for In Vitro<br>Diagnostic Use Only for the<br>quantitative measurement of<br>sex hormone binding globulin<br>(SHBG) in human serum or<br>Na heparinized plasma on<br>Tosoh AIA System Analyzers.<br>The ST AIA-PACK SHBG<br>assay is intended for use as<br>an aid in the diagnosis of<br>androgen disorders. | The ARCHITECTTM SHBG<br>assay is a chemiluminescent<br>microparticle immunoassay<br>(CMIA) for the quantitative<br>determination of sex hormone<br>binding globulin (SHBG) in<br>human serum and plasma on<br>the ARCHITECT iSystem.<br>The ARCHITECT SHBG<br>assay is intended for use as<br>an aid in the diagnosis of<br>androgen disorders. |
| Specimen type | Serum or sodium heparinized<br>plasma | Human serum or plasma<br>(Lithium Heparin, Sodium<br>Heparin, Ammonium Heparin,<br>Potassium EDTA) |
| Interference | No interference from:<br>Hemoglobin<br>Free bilirubin<br>Conjugated bilirubin<br>Lipemia, as indicated by<br>triglyceride concentration<br>Protein, as indicated by<br>human albumin concentration<br>HAMA | Non-significant interferences<br>with:<br>Hemoglobin<br>Bilirubin<br>Triglycerides<br>Protein |
| Limit of detection | 0.02 nmol/L | 0.02 nmol/L |
| Specificity/Cross Reactivity | Non-cross reactive:<br>Alpha-Fetoprotein (AFP)<br>Estradiol<br>Thyroxin-Binding Globulin<br>Transferrin<br>11-Deoxycortisol<br>5alpha-Dihydroxytestosterone<br>Cortisol<br>Testosterone<br>Thyroglobulin | Non-cross reactive:<br>AFP<br>Estradiol<br>Thyroxin-Binding Globulin<br>Transferrin<br>11-Deoxycortisol<br>5alpha-Dihydroxytestosterone<br>Cortisol<br>Testosterone<br>Thyroglobulin |
| Precision | Within-run: <10% CV from<br>15.5 -175.6 nmol/L | Within-run: <10% CV from<br>16.9 - 147.9 nmol/L |
| | Total Precision <10% CV from<br>15.5 – 175.6 nmol/L | Total Precision: <10% CV from 16.9 - 147.9 nmol/L |
| Hook Effect | The "hook effect"<br>phenomenon may occur only<br>at SHBG concentrations ><br>10,000 nmol/L. | The "hook effect"<br>phenomenon may occur only<br>at SHBG concentrations ><br>10,000 nmol/L. |

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## Differences

| Parameter | ST AIA-PACK SHBG | Abbott Architect SHBG Kit<br>(k060818) |
|------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Test Methodology | Fluorescence Immunosassay | Chemiluminescent<br>Microparticle<br>Immunoassay (CMIA) |
| Components | Unit dose test cups<br>containing twelve lyophilized<br>magnetic beads coated with<br>anti-SHBG mouse<br>monoclonal antibody and 100<br>µL of anti-SHBG mouse<br>monoclonal antibody<br>conjugated to bovine alkaline<br>phosphatase with sodium<br>azide as a preservative. | Microparticles<br>1 or 4 Bottle(s) (6.6 ml each)<br>Anti-SHBG(mouse<br>monoclonal) coated<br>microparticles in TRIS buffer.<br>Preservative: sodium azide |
| Specificity/Cross Reactivity | Non cross reactive:<br>Plasminogen<br>TSH<br>Fibrinogen<br>Human IgA<br>Human IgG<br>Corticosteroid Binding<br>Globulin | Not specified |
| Expected Values | 488 samples - 244 male and<br>244 female<br>Male (21 to 49 years old) 10 -<br>68 nmol/L<br>Male (≥50 years old)16-125<br>nmol/L<br>Females (pre-menopausal)<br>18 – 260 nmol/L<br>Females (post-menopausal)<br>15 - 185 nmol/L | 319 samples - 152 male and<br>167 female<br>Male 11.2 - 78.1 nmol/L<br>Female 11.7 - 137.2 nmol/L |
| Interference | No interference from:<br>Ascorbic acid<br>Heparin | Not specified or tested |
| Linearity (measuring range) | 0.2 to 250 nmol/L. | 0.1 to 250 nmol/L |

# Similarities

# Calibrator Set

| Parameter | ST AIA-PACK SHBG<br>Calibrator Set | Architect SHBG Calibrator<br>Kit (k060818) |
|--------------|------------------------------------|--------------------------------------------|
| Intended use | The ST AIA-PACK SHBG | The ARCHITECT "" SHBG |

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| | Calibrator Set is intended for<br>In Vitro Diagnostic Use Only<br>for the calibration of the ST<br>AIA-PACK SHBG assay. | Calibrators are for the<br>calibration of the<br>ARCHITECT iSystem when<br>used for the quantitative<br>determination of SHBG in<br>human serum and plasma. |
|-----------------------|-------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Number of Calibrators | 6 | 6 |

## Differences

## Calibrator Set

| Parameter | AIA-PACK SHBG Calibrator<br>Set | Architect SHBG Calibrator<br>Kit (k060818) |
|------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------|
| Components | Bovine serum with assigned<br>levels of sex hormone binding<br>globulin (SHBG). | SHBG (human, purified) in<br>phosphate buffered saline<br>with protein (goat) stabilizer |
| Traceability/Standardization | Traceable to the 2nd<br>International Standard for<br>SHBG from the National<br>Institute for Biological<br>Standards and Control<br>(NIBSC) code 08/266. | Traceable to the WHO<br>Standard Material NIBSC<br>CODE: 95/560. |

# PERFORMANCE CHARACTERISTICS

# Precision

The precision study was developed with reference to the CLSI protocol entitled: Evaluation of Precision Performance of Quantitative Measurement Methods (EP5-A2).

The precision study for the ST AIA-PACK SHBG assay was evaluated utilizing three AIA-2000 analyzers and three different lots of reagents. Precision was assessed by assaying three levels of serum and heparinized plasma specimens. Estimates of total and within run precision were obtained from measurements of 2 replicates in a single run, 2 times a day for 20 non-consecutive days. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV). In addition, all of the data were combined to assess the within run, between run, between lot and total precision.

## Within run precision

| | Mean | Pooled SD | CV |
|----------|----------|-----------|-----|
| Sample | (nmol/L) | (nmol/L) | (%) |
| Serum 1L | 17.3 | 0.454 | 2.6 |
| Serum 1M | 52.7 | 1.48 | 2.8 |
| Serum 1H | 154.9 | 4.96 | 3.2 |
| Serum 2L | 18.0 | 0.39 | 2.1 |
| Serum 2M | 54.8 | 1.29 | 2.3 |
| Serum 2H | 158.9 | 4.07 | 2.6 |
| Serum 3L | 18.8 | 0.56 | 2.9 |

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| Serum 3M | 58.4 | 1.40 | 2.4 |
|-----------------------|-------|------|-----|
| Serum 3H | 175.6 | 5.26 | 3.0 |
| Heparinized Plasma 1L | 15.5 | 0.48 | 3.1 |
| Heparinized Plasma 1M | 62.9 | 1.70 | 2.7 |
| Heparinized Plasma 1H | 139.6 | 3.92 | 2.8 |
| Heparinized Plasma 2L | 16.1 | 0.39 | 2.4 |
| Heparinized Plasma 2M | 65.2 | 1.03 | 1.6 |
| Heparinized Plasma 2H | 142.2 | 3.70 | 2.6 |
| Heparinized Plasma 3L | 17.2 | 0.43 | 2.5 |
| Heparinized Plasma 3M | 70.7 | 1.59 | 2.3 |
| Heparinized Plasma 3H | 159.3 | 4.90 | 3.1 |

# Total Precision

| | Mean | Pooled SD | CV |
|-----------------------|----------|-----------|-----|
| Sample | (nmol/L) | (nmol/L) | (%) |
| Serum 1L | 17.3 | 0.485 | 2.8 |
| Serum 1M | 52.7 | 1.62 | 3.1 |
| Serum 1H | 154.9 | 5.13 | 3.3 |
| Serum 2L | 18.0 | 0.54 | 3.0 |
| Serum 2M | 54.8 | 1.75 | 3.2 |
| Serum 2H | 158.9 | 4.80 | 3.0 |
| Serum 3L | 18.8 | 0.61 | 3.2 |
| Serum 3M | 58.4 | 1.51 | 2.6 |
| Serum 3H | 175.6 | 6.43 | 3.7 |
| Heparinized Plasma 1L | 15.5 | 0.54 | 3.5 |
| Heparinized Plasma 1M | 62.9 | 2.03 | 3.2 |
| Heparinized Plasma 1H | 139.6 | 5.13 | 3.7 |
| Heparinized Plasma 2L | 16.1 | 0.52 | 3.2 |
| Heparinized Plasma 2M | 65.2 | 1.55 | 2.4 |
| Heparinized Plasma 2H | 142.2 | 4.39 | 3.1 |
| Heparinized Plasma 3L | 17.2 | 0.52 | 3.0 |
| Heparinized Plasma 3M | 70.7 | 1.98 | 2.8 |
| Heparinized Plasma 3H | 159.3 | 5.42 | 3.4 |

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# Between run precision

| | Mean | Pooled SD | CV |
|-----------------------|----------|-----------|-----|
| Sample | (nmol/L) | (nmol/L) | (%) |
| Serum 1L | 17.3 | 0.33 | 1.9 |
| Serum 1M | 52.7 | 1.17 | 2.2 |
| Serum 1H | 154.9 | 3.75 | 2.4 |
| Serum 2L | 18.0 | 0.37 | 2.0 |
| Serum 2M | 54.8 | 1.32 | 2.4 |
| Serum 2H | 158.9 | 3.39 | 2.1 |
| Serum 3L | 18.8 | 0.43 | 2.3 |
| Serum 3M | 58.4 | 1.14 | 2.0 |
| Serum 3H | 175.6 | 4.99 | 2.8 |
| Heparinized Plasma 1L | 15.5 | 0.42 | 2.7 |
| Heparinized Plasma 1M | 62.9 | 1.63 | 2.6 |
| Heparinized Plasma 1H | 139.6 | 4.29 | 3.1 |
| Heparinized Plasma 2L | 16.1 | 0.41 | 2.5 |
| Heparinized Plasma 2M | 65.2 | 0.95 | 1.5 |
| Heparinized Plasma 2H | 142.2 | 2.88 | 2.0 |
| Heparinized Plasma 3L | 17.2 | 0.42 | 2.4 |
| Heparinized Plasma 3M | 70.7 | 1.40 | 2.0 |
| Heparinized Plasma 3H | 159.3 | 4.16 | 2.6 |

# Between day precision

| | Mean | Pooled SD | CV |
|-----------------------|----------|-----------|-----|
| Sample | (nmol/L) | (nmol/L) | (%) |
| Serum 1L | 17.3 | 0.28 | 1.6 |
| Serum 1M | 52.7 | 0.91 | 1.7 |
| Serum 1H | 154.9 | 2.59 | 1.7 |
| Serum 2L | 18 | 0.39 | 2.1 |
| Serum 2M | 54.8 | 1.29 | 2.3 |
| Serum 2H | 158.9 | 4.07 | 2.6 |
| Serum 3L | 18.8 | 0.56 | 2.9 |
| Serum 3M | 58.4 | 1.4 | 2.4 |
| Serum 3H | 175.6 | 5.26 | 3.0 |
| Heparinized Plasma 1L | 15.5 | 0.48 | 3.1 |
| Heparinized Plasma 1M | 62.9 | 1.7 | 2.7 |
| Heparinized Plasma 1H | 139.6 | 3.92 | 2.8 |
| Heparinized Plasma 2L | 16.1 | 0.39 | 2.4 |
| Heparinized Plasma 2M | 65.2 | 1.03 | 1.6 |
| Heparinized Plasma 2H | 142.2 | 3.7 | 2.6 |
| Heparinized Plasma 3L | 17.2 | 0.43 | 2.5 |

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| Heparinized Plasma 3M | 70.7 | 1.59 | 2.3 |
|-----------------------|-------|------|-----|
| Heparinized Plasma 3H | 159.3 | 4.9 | 3.1 |

## Representative Lot-to-Lot Precision

| Sample | Mean<br>(n=80)<br>nmol/L | Within-Run | | Between-Run | | Total | |
|-------------------|--------------------------|------------|-----|-------------|-----|-------|-----|
| | | SD | %CV | SD | %CV | SD | %CV |
| Serum Low | 18.0 | 0.39 | 2.1 | 0.37 | 2.0 | 0.53 | 3.0 |
| Serum medium | 54.8 | 1.29 | 2.3 | 1.32 | 2.4 | 1.8 | 3.3 |
| Serum High | 158.9 | 4.07 | 2.6 | 3.39 | 2.1 | 4.93 | 3.1 |
| Hep Plasma Low | 16.1 | 0.4 | 2.4 | 0.4 | 2.5 | 0.6 | 3.4 |
| Hep Plasma Medium | 65.2 | 1.0 | 1.6 | 1.0 | 1.5 | 1.5 | 2.3 |
| Hep Plasma High | 142.2 | 3.7 | 2.6 | 2.9 | 2.0 | 4.4 | 3.1 |

## Combined Summary Table

| Sample | | Serum-<br>1 | Serum-<br>2 | Serum-<br>3 | Hep<br>Plasma-<br>1 | Hep<br>Plasma-<br>2 | Hep<br>Plasma-<br>3 |
|------------------------|-----|-------------|-------------|-------------|---------------------|---------------------|---------------------|
| Mean Conc.<br>(nmol/L) | | 18.1 | 55.3 | 163.1 | 16.3 | 66.3 | 147.0 |
| Within | SD | 0.47 | 1.39 | 4.79 | 0.43 | 1.47 | 4.21 |
| Run | %CV | 2.6 | 2.5 | 2.9 | 2.7 | 2.2 | 2.9 |
| Between | SD | 0.38 | 1.21 | 4.10 | 0.42 | 1.36 | 3.83 |
| Run | %CV | 2.1 | 2.2 | 2.5 | 2.6 | 2.0 | 2.6 |
| Between | SD | 0.72 | 2.57 | 9.59 | 0.74 | 3.47 | 9.19 |
| Day | %CV | 4.0 | 4.6 | 5.9 | 4.6 | 5.2 | 6.3 |
| Between | SD | 0.77 | 1.54 | 2.77 | 0.84 | 1.59 | 1.79 |
| Lot | %CV | 4.3 | 2.8 | 1.7 | 5.1 | 2.4 | 1.2 |
| | SD | 0.84 | 2.88 | 9.42 | 0.85 | 3.75 | 10.04 |
| Total | %CV | 4.6 | 5.2 | 5.8 | 5.3 | 5.7 | 6.8 |

## Linearity:

Linearity: The linearity for ST AIA-PACK SHBG was determined, based on guidance from CLSI Protocol EP6-A entitled: Evaluation of the Linearity of Quantitative Measurement Procedures: a Statistical Approach: Approved Guideline. The linearity was measured on the AIA-2000 instrument and has been demonstrated to be linear from 0.2 to 250 nmol/L.

## Correlation

The methods comparison study was based on guidance from EP9-A2.

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## Method Comparison

A total of 126 serum specimens were assayed in singleton utilizing the ST AIA-PACK SHBG assay on the AIA-2000 analyzer and the predicate SHBG.

| | Deming | Regular |
|----------------|------------------------------------------|--------------------------------------------|
| Slope: | 0.949 (0.926 to 0.972) | 0.940 (0.917 to 0.964) |
| Intercept: | -0.64 (-2.61 to 1.34) | -0.09 (-2.06 to 1.89) |
| Corr Coef (R): | 0.991 | |
| Result Ranges: | Tosoh ST AIA-PACK SHBG<br>Predicate SHBG | 0.6 to 241.0 nmol/L<br>0.5 to 221.6 nmol/L |

*95% Confidence Intervals are shown in parentheses

#### Matrix Comparison

The correlation between serum (x) and Na heparinized plasma (y) on ST AIA-PACK SHBG was carried out using 116 patient specimens were diluted to obtain the low end of the measuring range.

| | Deming | Regular |
|----------------|----------------------------------|--------------------------------------------|
| Slope: | 0.977 (0.964 to 0.991) | 0.975 (0.961 to 0.988) |
| Intercept: | 0.269 (-0.629 to 1.168) | 0.415 (-0.483 to 1.313) |
| Corr Coef (R): | 0.997 | |
| Result Ranges | Serum:<br>Na heparinized plasma: | 0.2 to 219.1 nmol/L<br>0.2 to 219.9 nmol/L |

95% Confidence Intervals are shown in parentheses

## Specificity

The following substances were tested for cross-reactivity. Cross-reactivity is the percentage of the compound which will be identified as SHBG.

| Substance | Concentration<br>added | Cross-reactivity<br>(%) |
|------------------------------|------------------------|-------------------------|
| Alpha-Fetoprotein (AFP) | 48.4 $\mu$ g/dL | N.D. |
| Cortisol | 100 $\mu$ g/mL | 0.003 |
| 11-Deoxycortisol | 4 $\mu$ g/mL | 0.114 |
| 5alpha-Dihydroxytestosterone | 20 $\mu$ g/mL | 0.007 |
| Estradiol | 3600 pg/mL | N.D. |
| Testosterone | 20 $\mu$ g/mL | 0.019 |
| Thyroglobulin | 300 $\mu$ g/mL | 2.544 |
| Thyroxin-Binding Globulin | 200 $\mu$ g/mL | N.D. |
| Transferrin | 4 mg/mL | N.D. |
| Fibrinogen | 4.5 g/L | 0.120 |
| Plasminogen | 250 mg/L | N.D. |
| Human IgA | 367 mg/dL | 0.072 |
| Human IgG | 335 mg/dL | 0.218 |
| CBG | 35 mg/dL | 0.012 |
| TSH | 180 mIU/L | N.D. |

(N.D.: not detectable)

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## Reference Ranges

The reference range study was conducted with reference to the CLSI protocol entitled: How to Define and Determine Reference Intervals in the Clinical Laboratory (C28-A3) entitled: Defining, Establishing, and verifying Reference Intervals in the Clinical Laboratory; Approved Guideline -Third Edition.

The interval given here was determined in serum samples from 488 apparently healthy American and European individuals.

| | n | Range (nmol/L) |
|--------------------------------------------|-----|----------------|
| Male (21 to 49 years old) | 121 | 10 - 68 |
| Male (≥50 years old) | 123 | 16 - 125 |
| Females (pre-menopausal ≥ 21 years if age) | 122 | 18 - 260 |
| Females (post-menopausal) | 122 | 15 - 185 |

#### Interference

Interference is defined, for the purposes of this study, with recovery outside of 10 % of the known concentration of the specimen after the following substances are added to human specimens.

- Hemoglobin (up to 446 mg/dL), free bilirubin (up to 17.6 mg/dL), and conjugated bilirubin ● (up to 18.5 mq/dL) do not interfere with the assay.
- Lipemia, as indicated by triglyceride concentration (up to 1,667 mg/dL), does not interfere with the assay.
- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. .
- Protein, as indicated by human albumin concentration (up to 5,00 g/dL added to samples ● from apparently healthy subjects), does not interfere with the assay.
- Na Heparin (up to 100.0 U/mL) does not interfere with the assay. ●
- . Rheumatoid factor (up to 550 IU/mL) does not interfere with the assay.
- . HAMA (up to 24,269 ng/mL) does not interfere with the assay.

## Limit of Detection (LoD) and Limit of Quantitation (LoQ):

The LoD and LoQ for ST AIA-PACK SHBG was determined, according to CLSI quideline EP17-A entitled: Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline.

The LoB was determined by 60 measurements of 4 different blank specimens. The LoB was the value at the 95th percentile. In this case LoB was determined to be 0.017 nmol/L.

The LoD was determined by 12 measurements of 10 low level sample range was chosen to be between LoB and 5XLoB. The LoD was determined to be 0.063 nmol/L.

The Limit of Quantification (LoQ) 12 measurements of 5 samples for ST AIA-PACK SHBG. The test results from the LoD study were used to calculate the %TE at that level. The LoQ was determined to be 0.2 nmol/L based on a % total error of 12%.

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## Calibrator Stability

#### Real Time Testing

ST AIA-PACK SHBG Calibrator Set were stored at refrigerated temperatures and assayed at 3, 6, 9, 12 and 13 months after the day of the first assay.

The acceptance criteria for recovery was within 100 +/- 10%.

The criterion for reproducibility (CV %) was </= 10%.

Current real time studies support a 12 month shelf life at refrigerated temperatures from the date of manufacturing.

## Open Vial Stability

Open vial stability of the ST AIA-PACK SHBG Calibrator Set was assessed by reconstituting the material according to the package insert. Samples were reconstituted and stored at refrigerated temperatures for 2 days and tested for SHBG.

The criterion for recovery was within 100 +/-10%.

The criterion for reproducibility (CV %) was </= 10%.

Current open vial studies support an in-use claim of 1 day when refrigerated.

## Summary of Calibrator Value Assignment

The value assignment of the ST AIA-PACK SHBG Calibrator Set is determined on a lot-by-lot basis and is designed to provide an assay calibration range of 0.005 to 12.5 nmol/L of SHBG. The calibrators in this set are referenced to the 2nd International Standard for SHBG from the National Institute for Biological Standards and Control (NIBSC) code 08/266.

The primary reference material was prepared by diluting the SHBG with the calibrator base. The value of SHBG was assigned based on WHO 2nd IS SHBG using ST AIA-PACK SHBG

The value of the secondary reference material was assigned using the AIA instruments with the primary reference material as calibrator. The value was verified by comparing measured results with those obtained with the previous lot for patient samples.

Note: As each manufactured calibrator concentration corresponds to a unique rate generated by the reaction detected by the instrument, the rate generated by the patient sample (which is diluted 1:20) would correspond to a rate on the curve generated by the manufactured calibrator. The value would then be multiplied automatically by the factor of 20 to generate the actual concentration of SHBG in the sample.

#### Standards:

| Number | FDA<br>Recognition<br>Number | Revision<br>Date | Title |
|--------|------------------------------|------------------|----------------------------------------------------------------------------------------------------------------------|
| EP5-A2 | 7-110 | 10/31/2005 | Evaluation of Precision Performance of<br>Quantitative Measurement Methods; Approved<br>Guideline-Second Edition |
| EP6-A | 7-193 | 03/18/2009 | Evaluation of the Linearity of Quantitative<br>Measurement Procedures: a Statistical<br>Approach: Approved Guideline |

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| EP28-A3c | 7-224 | 10/1/2010 | Defining, Establishing, and verifying Reference<br>Intervals in the Clinical Laboratory; Approved<br>Guideline – Third Edition |
|----------|-------|------------|--------------------------------------------------------------------------------------------------------------------------------|
| EP07-A2 | 7-127 | 05/21/2007 | Interference Testing in Clinical Chemistry;<br>Approved Guideline - Second Edition |
| EP9-A2 | 7-92 | 07/01/2010 | Method Comparison and Bias Estimation Using<br>Patient Samples; Approved Guideline-Second<br>Edition |

## Conclusion:

The Tosoh Bioscience, Inc. ST AIA-PACK SHBG is substantially equivalent to the Abbott Architect SHBG (k)060818 for In Vitro Diagnostic Use Only for the quantitative measurement of SHBG in human serum or Na heparinized plasma on Tosoh AIA System Analyzers. The ST AIA-PACK SHBG assay is intended for use as an aid in the diagnosis of androgen disorders.