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 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 magnetic 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.

AI/ML Overview

The provided text describes the performance characteristics of the ST AIA-PACK SHBG device, which is an in vitro diagnostic assay for measuring Sex Hormone Binding Globulin (SHBG). The studies conducted for this device do not involve AI or human readers for image interpretation, but rather focus on analytical performance criteria typical for a laboratory diagnostic assay. Therefore, questions related to AI assistance, human reader improvement, and adjudication methods are not applicable.

Here's an analysis of the acceptance criteria and study information provided:

1. Table of Acceptance Criteria and Reported Device Performance

For analytical devices like the ST AIA-PACK SHBG, acceptance criteria are typically specified in terms of performance metrics like precision (CV%), linearity (measuring range), limit of detection (LoD), and correlation with a predicate device. The document generally presents the performance metrics achieved by the device without explicitly stating pre-defined "acceptance criteria" numerical targets. However, comparisons to the predicate device and CLSI guidelines serve as implicit benchmarks for acceptability.

Performance Characteristic	Implicit Acceptance Criteria (based on CLSI guidelines/predicate comparison/industry standards)	Reported Device Performance (ST AIA-PACK SHBG)
Precision (Total Precision CV%)	Typically <10% for immunoassay (Often below 5-10% for various levels)	Serum: 2.8% to 3.7% (for individual lots/levels), Combined Summary Table: 4.6% to 5.8% (for serum), Plasma: 3.5% to 3.7% (for individual lots/levels), Combined Summary Table: 5.3% to 6.8% (for plasma). All reported values are within the typical acceptance range for immunoassays.
Linearity (Measuring Range)	Should cover a clinically relevant range. Predicate range: 0.1 to 250 nmol/L.	Demonstrated linearity from 0.2 to 250 nmol/L. This range is comparable to the predicate and clinically relevant.
Limit of Detection (LoD)	Low enough for clinical utility. Predicate LoD: 0.02 nmol/L.	0.063 nmol/L. The predicate specified a LoD of 0.02 nmol/L, meaning the subject device's LoD is slightly higher, but still low.
Limit of Quantitation (LoQ)	Low enough for accurate quantitation.	0.2 nmol/L (based on 12% total error).
Method Comparison (Correlation with Predicate)	High correlation (e.g., Correlation Coefficient (R) > 0.95, slope near 1, intercept near 0).	Deming Slope: 0.949 (0.926 to 0.972), Intercept: -0.64 (-2.61 to 1.34), Corr Coef (R): 0.991. These values demonstrate excellent correlation with the predicate.
Matrix Comparison (Serum vs. Plasma)	High correlation between different sample matrices.	Deming Slope: 0.977 (0.964 to 0.991), Intercept: 0.269 (-0.629 to 1.168), Corr Coef (R): 0.997. These values demonstrate excellent correlation between serum and Na heparinized plasma.
Interference	No significant interference from common interfering substances (e.g., hemoglobin, bilirubin, lipids, HAMA). Recovery typically within +/-10%.	No interference observed from: Hemoglobin (up to 446 mg/dL), free and conjugated bilirubin (up to 17.6 mg/dL and 18.5 mg/dL respectively), lipemia (triglycerides up to 1,667 mg/dL), ascorbic acid (up to 20 mg/dL), protein (albumin up to 5.00 g/dL), Na Heparin (up to 100.0 U/mL), Rheumatoid factor (up to 550 IU/mL), HAMA (up to 24,269 ng/mL). This meets the non-interference expectation.
Cross-Reactivity	Minimal to no cross-reactivity with related or common substances.	Generally N.D. (not detectable) or very low percentage (e.g., 0.003% for Cortisol, 0.019% for Testosterone). Thyroglobulin had a 2.544% cross-reactivity, which may be noted but specific acceptance for such levels is not provided.
Calibrator Stability (Real Time)	Recovery within 100 +/- 10%; Reproducibility (CV%) <= 10%.	Supports 12-month shelf life at refrigerated temperatures.
Calibrator Stability (Open Vial)	Recovery within 100 +/- 10%; Reproducibility (CV%) <= 10%.	Supports 1-day in-use claim when refrigerated.

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

Precision Study:
- Three levels of serum and heparinized plasma specimens were assayed.
- Measurements were taken in 2 replicates in a single run, 2 times a day for 20 non-consecutive days.
- This implies a total of 3 (levels) * 2 (matrices) * 2 (reps/run) * 2 (runs/day) * 20 (days) = 480 measurements per reagent lot, but the actual number of distinct biological samples is not directly specified beyond "three levels of serum and heparinized plasma specimens".
Linearity Study: Sample size not explicitly stated for this particular study, but it was performed with guidance from CLSI Protocol EP6-A.
Method Comparison: 126 serum specimens.
Matrix Comparison: 116 patient specimens (diluted to obtain the low end of the measuring range).
Reference Ranges Study: 488 apparently healthy American and European individuals (244 male and 244 female).
Interference Study: Not specified, but involved adding various interfering substances to "human specimens".
Limit of Detection (LoD) and Limit of Quantitation (LoQ):
- LoD: 60 measurements of 4 different blank specimens; 12 measurements of 10 low-level samples.
- LoQ: 12 measurements of 5 samples.

Data Provenance:

Reference Ranges: "488 apparently healthy American and European individuals." This indicates a prospective collection from multiple geographical locations.
For other studies (Precision, Method Comparison, Matrix Comparison, Interference, LoD/LoQ), the provenance (country of origin, retrospective/prospective) is not explicitly stated, but the use of "human serum or Na heparinized plasma," "patient specimens," and "human specimens" suggests clinical samples.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts

This information is not applicable as the device is an in vitro diagnostic assay that generates quantitative measurements of a biochemical marker (SHBG). The "ground truth" for such devices is established through analytical validation against reference methods or accepted standards, not through expert consensus on qualitative assessments like image interpretation. There are no "experts" in the traditional sense mentioned, but rather scientific and clinical validation methodologies.

4. Adjudication Method for the Test Set

This information is not applicable for the same reasons as in point 3. Adjudication is typically relevant for studies where subjective interpretation (e.g., by radiologists) is part of establishing ground truth or evaluating performance, which is not the case for this quantitative assay.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, If So, What Was the Effect Size of How Much Human Readers Improve with AI vs. Without AI Assistance

This information is not applicable. This device is a standalone laboratory assay and does not involve AI assistance or human readers for diagnostic interpretation in the manner described for MRMC studies (e.g., in radiology).

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done

Yes, the studies performed (e.g., precision, linearity, LoD, method comparison) represent the standalone performance of the analytical instrument and assay kit. These tests evaluate the accuracy and reliability of the device's measurements directly, without human interpretation in the diagnostic pathway, other than performing the test and reviewing the quantitative results.

7. The Type of Ground Truth Used

The ground truth used for this device's validation is primarily reference methods and established analytical standards:

Predicate Device Comparison: The ST AIA-PACK SHBG assay's performance was compared against the Abbott Architect SHBG Immunoassay (K060818), which serves as a legally marketed predicate and a standard for comparison.
International Standards: The calibrators are traceable to the 2nd International Standard for SHBG from the National Institute for Biological Standards and Control (NIBSC) code 08/266. This is a critical aspect of establishing the "ground truth" for the quantitative values.
CLSI Protocols: The studies were conducted with reference to widely accepted Clinical and Laboratory Standards Institute (CLSI) protocols (e.g., EP5-A2 for precision, EP6-A for linearity, EP9-A2 for method comparison), which define rigorous scientific methods for demonstrating analytical performance.

8. The Sample Size for the Training Set

The document does not describe a "training set" in the context of machine learning or AI, as this is an analytical diagnostic assay. The term "training set" would not typically apply here. Instead, numerous samples are used during the development and validation phases to establish the assay's performance characteristics. For example, during the creation of calibrators or development of the assay, various characterized samples would have been used.

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

As there is no "training set" in the context of AI/machine learning for this device, this question is not applicable. The ground truth for the assay's performance validation is established through comparison to predicate devices, traceability to international standards, and adherence to CLSI guidelines, as described in point 7.

Summary

{0}------------------------------------------------

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.

{1}------------------------------------------------

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 (800) 638 2041 or (301) 796-7100 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 (800) 638-2041 or (301) 796-7100 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

{2}------------------------------------------------

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)

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)
Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

{3}------------------------------------------------

510(k) Summary

{4}------------------------------------------------

510(k) Summary

K143075

ST AIA-PACK SHBG

Date:	June 30, 2015
Submitter:	Tosoh Bioscience, Inc3600 Gantz RoadGrove City, OH 43123
Contact Person:	Robert L. WickRegulatory Specialist6000 Shoreline Ct., Ste. 101South San Francisco, CA 94080
	Phone: 650-636-8117
	Fax: 650-636-8121
	Email: Robert.Wick@tosoh.com
Device Name:	ST AIA-PACK SHBG
Classification:	Class I, reservedCDZClinical Chemistry21 CFR 862.1680
Device Name:	ST AIA-PACK SHBG Calibrator Set
Classification	Class IIJITClinical Chemistry21 CFR 862.1150
Predicate Device:	K060818Abbott/ BIOKIT S.A.ARCHITECT SHBG REAGENT KIT, ARCHITECTSHBG CALIBRATOR KIT

{5}------------------------------------------------

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:

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.

{6}------------------------------------------------

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(K060818)
Intended use	For the quantitativemeasurement of SHBG inhuman serum and plasma.	For the quantitativemeasurement of SHBG inhuman serum and plasma.
Indications for Use	ST AIA-PACK SHBG isdesigned for In VitroDiagnostic Use Only for thequantitative measurement ofsex hormone binding globulin(SHBG) in human serum orNa heparinized plasma onTosoh AIA System Analyzers.The ST AIA-PACK SHBGassay is intended for use asan aid in the diagnosis ofandrogen disorders.	The ARCHITECTTM SHBGassay is a chemiluminescentmicroparticle immunoassay(CMIA) for the quantitativedetermination of sex hormonebinding globulin (SHBG) inhuman serum and plasma onthe ARCHITECT iSystem.The ARCHITECT SHBGassay is intended for use asan aid in the diagnosis ofandrogen disorders.
Specimen type	Serum or sodium heparinizedplasma	Human serum or plasma(Lithium Heparin, SodiumHeparin, Ammonium Heparin,Potassium EDTA)
Interference	No interference from:HemoglobinFree bilirubinConjugated bilirubinLipemia, as indicated bytriglyceride concentrationProtein, as indicated byhuman albumin concentrationHAMA	Non-significant interferenceswith:HemoglobinBilirubinTriglyceridesProtein
Limit of detection	0.02 nmol/L	0.02 nmol/L
Specificity/Cross Reactivity	Non-cross reactive:Alpha-Fetoprotein (AFP)EstradiolThyroxin-Binding GlobulinTransferrin11-Deoxycortisol5alpha-DihydroxytestosteroneCortisolTestosteroneThyroglobulin	Non-cross reactive:AFPEstradiolThyroxin-Binding GlobulinTransferrin11-Deoxycortisol5alpha-DihydroxytestosteroneCortisolTestosteroneThyroglobulin
Precision	Within-run: <10% CV from15.5 -175.6 nmol/L	Within-run: <10% CV from16.9 - 147.9 nmol/L
	Total Precision <10% CV from15.5 – 175.6 nmol/L	Total Precision: <10% CV from 16.9 - 147.9 nmol/L
Hook Effect	The "hook effect"phenomenon may occur onlyat SHBG concentrations >10,000 nmol/L.	The "hook effect"phenomenon may occur onlyat SHBG concentrations >10,000 nmol/L.

{7}------------------------------------------------

Differences

Parameter	ST AIA-PACK SHBG	Abbott Architect SHBG Kit(K060818)
Test Methodology	Fluorescence Immunosassay	ChemiluminescentMicroparticleImmunoassay (CMIA)
Components	Unit dose test cupscontaining twelve lyophilizedmagnetic beads coated withanti-SHBG mousemonoclonal antibody and 100µL of anti-SHBG mousemonoclonal antibodyconjugated to bovine alkalinephosphatase with sodiumazide as a preservative.	Microparticles1 or 4 Bottle(s) (6.6 ml each)Anti-SHBG(mousemonoclonal) coatedmicroparticles in TRIS buffer.Preservative: sodium azide
Specificity/Cross Reactivity	Non cross reactive:PlasminogenTSHFibrinogenHuman IgAHuman IgGCorticosteroid BindingGlobulin	Not specified
Expected Values	488 samples - 244 male and244 femaleMale (21 to 49 years old) 10 -68 nmol/LMale (≥50 years old)16-125nmol/LFemales (pre-menopausal)18 – 260 nmol/LFemales (post-menopausal)15 - 185 nmol/L	319 samples - 152 male and167 femaleMale 11.2 - 78.1 nmol/LFemale 11.7 - 137.2 nmol/L
Interference	No interference from:Ascorbic acidHeparin	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 SHBGCalibrator Set	Architect SHBG CalibratorKit (K060818)
Intended use	The ST AIA-PACK SHBG	The ARCHITECT "" SHBG

{8}------------------------------------------------

	Calibrator Set is intended forIn Vitro Diagnostic Use Onlyfor the calibration of the STAIA-PACK SHBG assay.	Calibrators are for thecalibration of theARCHITECT iSystem whenused for the quantitativedetermination of SHBG inhuman serum and plasma.
Number of Calibrators	6	6

Differences

Calibrator Set

Parameter	AIA-PACK SHBG CalibratorSet	Architect SHBG CalibratorKit (K060818)
Components	Bovine serum with assignedlevels of sex hormone bindingglobulin (SHBG).	SHBG (human, purified) inphosphate buffered salinewith protein (goat) stabilizer
Traceability/Standardization	Traceable to the 2ndInternational Standard forSHBG from the NationalInstitute for BiologicalStandards and Control(NIBSC) code 08/266.	Traceable to the WHOStandard Material NIBSCCODE: 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

{9}------------------------------------------------

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

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

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

{11}------------------------------------------------

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(n=80)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-1	Serum-2	Serum-3	HepPlasma-1	HepPlasma-2	HepPlasma-3
Mean Conc.(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.

{12}------------------------------------------------

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 SHBGPredicate SHBG	0.6 to 241.0 nmol/L0.5 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:Na heparinized plasma:	0.2 to 219.1 nmol/L0.2 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	Concentrationadded	Cross-reactivity(%)
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)

{13}------------------------------------------------

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%.

{14}------------------------------------------------

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	FDARecognitionNumber	RevisionDate	Title
EP5-A2	7-110	10/31/2005	Evaluation of Precision Performance ofQuantitative Measurement Methods; ApprovedGuideline-Second Edition
EP6-A	7-193	03/18/2009	Evaluation of the Linearity of QuantitativeMeasurement Procedures: a StatisticalApproach: Approved Guideline

{15}------------------------------------------------

EP28-A3c	7-224	10/1/2010	Defining, Establishing, and verifying ReferenceIntervals in the Clinical Laboratory; ApprovedGuideline – Third Edition
EP07-A2	7-127	05/21/2007	Interference Testing in Clinical Chemistry;Approved Guideline - Second Edition
EP9-A2	7-92	07/01/2010	Method Comparison and Bias Estimation UsingPatient Samples; Approved Guideline-SecondEdition

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.

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.