The Optilite Freelite Mx Kappa Free Kit is intended for the quantitative in vitro measurement of Kappa free light chains in serum and urine using the Binding Site Optilite analyser. Measurement of free light chains aids in the diagnosis and monitoring of multiple myeloma, lymphocytic neoplasms, Waldenström's macroglobulinaemia, AL amyloidosis, light chain deposition disease and connective tissue diseases such as systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings.

The Optilite Freelite Mx Lambda Free Kit is intended for the quantitative in vitro measurement of Lambda free light chains in serum and urine using the Binding Site Optilite analyser. Measurement of free light chains aids in the diagnosis and monitoring of multiple myeloma, lymphocytic neoplasms, Waldenström's macroglobulinaemia, AL amyloidosis, light chain deposition disease and connective tissue diseases such as systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings.

Device Description

The Optilite Freelite Mx Kappa Free Kit is comprised of the following reagents: Latex Reagent: Sheep anti-Human Kappa (polyclonal monospecific) antibody (F(ab')2 fragment) bound to 200nm polystyrene latex particles. In 0.033M Glycine buffered saline pH8 (containing Sodium Azide (0.033%), Proclin (0.05%), Benzamidine (0.01%) and EACA (0.1%) as preservative plus BSA (0.033%)). Calibrator and Controls: Pooled human serum, supplied in stabilised liquid form. Containing 0.099% sodium azide, 0.1% EACA and 0.01% benzamidine as preservatives. Reaction Buffer: Containing 0.099% sodium azide as a preservative.

The Optilite Freelite Mx Lambda Free Kit is comprised of the following reagents: Latex Reagent: Sheep anti-Human Lambda (polyclonal monospecific) Free antibody (F(ab')2 fragment) bound to 200nm polystyrene latex particles. In 0.033M Glycine buffered saline pH8 (containing Sodium Azide (0.033%), Proclin (0.05%), Benzamidine (0.01%) and EACA (0.1%) as preservatives plus BSA (0.1665%)). Calibrator and Controls: Pooled human serum, supplied in stabilised liquid form. Containing 0.099% sodium azide, 0.1% EACA and 0.01% benzamidine as preservatives. Reaction Buffer: Containing 0.099% sodium azide as a preservative.

Note - In Optilite Freelite kits, the latex reagent and reaction buffer are supplied in a single wedge with a chamber for each fluid. They are therefore labelled as a single component Optilite Kappa Free Mx Reagent or Optilite Lambda Free Mx Reagent.

AI/ML Overview

The provided text describes the performance characteristics of the Optilite Freelite Mx Kappa Free Kit and Optilite Freelite Mx Lambda Free Kit, which are devices for the quantitative measurement of Kappa and Lambda free light chains in serum and urine.

Here's an analysis of the acceptance criteria and the study that proves the device meets these criteria, based on the provided text:

1. Table of Acceptance Criteria and Reported Device Performance

Since this is a submission for an in-vitro diagnostic device, the "acceptance criteria" are typically inferred from the performance targets set by the manufacturer and assessed against relevant CLSI (Clinical and Laboratory Standards Institute) guidelines. The reported device performance is directly stated in the tables within the document.

Precision/Reproducibility

Performance Metric	Acceptance Criteria (Inferred from CLSI Guidelines/Industry Standards for IVDs)	Reported Device Performance (Kappa Urine)	Reported Device Performance (Lambda Urine)
Kappa
Total %CV (Sample 1)	Typically < 10% (for low concentrations)	7.6%	-
Total %CV (Sample 2)	Typically < 10%	8.2%	-
Total %CV (Sample 3)	Typically < 10%	7.1%	-
Total %CV (Sample 4)	Typically < 10%	8.8%	-
Lambda
Total %CV (Sample 1)	Typically < 10% (for low concentrations)	-	7.3%
Total %CV (Sample 2)	Typically < 10%	-	8.7%
Total %CV (Sample 3)	Typically < 10%	-	8.2%
Total %CV (Sample 4)	Typically < 10%	-	8.1%
Total %CV (Sample 5)	Typically < 10%	-	8.6%

Linearity/Assay Reportable Range

Performance Metric	Acceptance Criteria (Inferred from CLSI EP6-A)	Reported Device Performance (Kappa)	Reported Device Performance (Lambda)
Linearity Range	Demonstrated across the intended measuring range with acceptable deviation	2.383 - 142.687 mg/L	3.87 - 160.02 mg/L
Deviation from Linearity	≤ 10%	≤ 10%	≤ 10%

Detection Limit

Performance Metric	Acceptance Criteria (Inferred from CLSI EP17-A)	Reported Device Performance (Kappa Serum)	Reported Device Performance (Kappa Urine)	Reported Device Performance (Lambda Serum)	Reported Device Performance (Lambda Urine)
Limit of Blank (LoB)	Accurately determined	0.195 mg/L	0.210 mg/L	0.285 mg/L	0.080 mg/L
Limit of Detection (LoD)	Accurately determined	0.224 mg/L	0.232 mg/L	0.393 mg/L	0.184 mg/L
Limit of Quantitation (LoQ)	Accurately determined and clinically relevant	0.330 mg/L	0.330 mg/L	0.740 mg/L	0.740 mg/L

Analytical Specificity (Interference)

Performance Metric	Acceptance Criteria (Inferred from CLSI EP7-A2)	Reported Device Performance (Kappa)	Reported Device Performance (Lambda)
Interference (% Deviation from Control)	≤ 10% for studied interferents	No significant interference observed for listed substances	No significant interference observed for listed substances

Method Comparison with Predicate Device

Performance Metric	Acceptance Criteria (Inferred for Substantial Equivalence)	Reported Device Performance (Kappa Free)	Reported Device Performance (Lambda Free)
Correlation Coefficient (r)	High correlation (typically > 0.95 or 0.975 for quantitative assays)	0.992	0.947
Regression Equation (y=mx+c)	Slope (m) close to 1, intercept (c) close to 0	y = 1.02x - 0.21	y = 1.11x - 0.27

Clinical Agreement (2x2 Table with Predicate)

Performance Metric	Acceptance Criteria (Inferred for Substantial Equivalence)	Reported Device Performance (Kappa)	Reported Device Performance (Lambda)
Positive Agreement	High, clinically acceptable	90.7% (82.7% to 95.2% CI)	73.7% (62.8% to 82.3% CI)
Negative Agreement	High, clinically acceptable	98.8% (93.3% to 99.8% CI)	100.0% (96.0% to 100.0% CI)
Overall Agreement	High, clinically acceptable	94.6% (90.0% to 97.1% CI)	88.1% (92.3% to 92.2% CI)

Notes on Acceptance Criteria: The document does not explicitly state quantitative acceptance criteria or pass/fail thresholds for many of these performance metrics. The criteria listed above are inferred based on typical expectations for in-vitro diagnostic devices undergoing 510(k) clearance, often guided by CLSI guidelines. The FDA's substantial equivalence determination implies that the reported performance is acceptable in comparison to the predicate device and the recognized standards.

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

Precision/Reproducibility:
- Kappa: Four urine sample pools were tested. The testing involved 21 days with two runs per day using one reagent lot on three analyzers.
- Lambda: Five urine sample pools were tested. The testing involved 23 days with two runs per day using one reagent lot on three analyzers.
- Provenance: Not explicitly stated, but likely in-house (Binding Site Group Ltd.) and conducted under controlled laboratory conditions, given it's part of analytical performance testing for an IVD.
Linearity/Assay Reportable Range:
- Sample Size: "serially diluted urine samples" were used. The exact number of individual samples or dilutions is not specified.
- Provenance: Not explicitly stated, but likely in-house.
Detection Limit:
- LoB: 60 determinations of blank samples (analyte depleted serum and urine at a low analyte concentration).
- LoQ: Five independent samples (serum and urine) tested twelve times over five days.
- Provenance: Not explicitly stated, but likely in-house.
Analytical Specificity (Interference):
- Sample Size: "a single urine sample for the Kappa Free assay and the Lambda Free assay". These were prepared with interferents.
- Provenance: Not explicitly stated, but likely in-house.
Method Comparison with Predicate Device:
- Kappa Free: 165 urine samples (including 86 with analyte levels below the reference limit).
- Lambda Free: 115 urine samples (including 59 with analyte levels below the reference limit).
- Provenance: Not explicitly stated, but these are patient samples tested against a predicate device. Given the context of a European manufacturer and the lack of specific geographical mention, they could be from various locations or a specific clinic associated with the manufacturer. Retrospective or prospective is not specified, but typically for method comparison, collected samples are used retrospectively.
Clinical Agreement (2x2 Table with Predicate):
- Kappa: 166 samples (derived from the 2x2 table: 78+1+8+79 = 166, though the sum of listed cells is 166, the total is given as 166, implying a discrepancy in the row/column totals). The method comparison stated 165, so there might be a rounding or slight difference in sample inclusion for the clinical agreement vs. regression.
- Lambda: 168 samples (derived from the 2x2 table totals). The method comparison stated 115. This suggests different sample sets or criteria for the 2x2 table analysis compared to the regression analysis, especially given the sample sizes below the reference limit quoted for the method comparison.
- Provenance: Not explicitly stated, but presumed to be from clinical settings for patient samples. These are likely retrospective samples previously analyzed by the predicate.

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

For an in-vitro diagnostic device like this, "ground truth" and "experts" are typically not established in the same way as for imaging devices requiring expert interpretation.

Ground Truth: For this device, the "ground truth" is typically the result obtained from a well-established, often reference, method or the predicate device that the new device is being compared against. In this case, the predicate device (Freelite® Human Kappa/Lambda Free Kit for use on the Siemens BN™II) serves as the comparator, from which the "Assay" results are compared.
Experts and Qualifications: There is no mention of external experts being used for establishing ground truth. The evaluation relies on analytical and clinical performance studies comparing the new device against the predicate or established analytical standards.

4. Adjudication Method (e.g., 2+1, 3+1, none) for the Test Set

Not applicable for this type of in-vitro diagnostic device performance study. Adjudication methods like 2+1 or 3+1 are common in studies where human readers interpret medical images and their interpretations need to be reconciled to establish a ground truth or a consensus opinion. For IVD devices, the "truth" for comparison is usually defined by the results of a reference method or legally marketed predicate.

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

Not applicable. MRMC studies, and improvement with AI assistance, are relevant to medical imaging devices where human readers (e.g., radiologists) interpret images. This submission is for an in-vitro diagnostic assay for laboratory measurement of biomarkers, which does not involve human interpretation of complex visual data in the same way.

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

This device is a standalone diagnostic assay (an "algorithm only" if one considers the assay chemistry and instrument processing as the "algorithm"). It performs quantitative measurements without direct human real-time interpretation influencing the numerical result. The human element is in collecting the sample, running the test, and interpreting the numerical result in a clinical context.

7. The Type of Ground Truth Used (expert consensus, pathology, outcomes data, etc.)

The ground truth for comparative studies and assessment of substantial equivalence is primarily established by:

Predicate Device Results: For method comparison and clinical agreement, the results from the legally marketed predicate device (Freelite® Human Kappa/Lambda Free Kit on the Siemens BN™II) are used as the reference ('ground truth') for comparison.
CLSI Guidelines: For analytical performance (precision, linearity, detection limits, interference), the "ground truth" is defined by adherence to the methodologies and performance expectations outlined in recognized CLSI guidelines (e.g., EP17-A, EP7-A2, EP6-A, EP5-A2).

8. The Sample Size for the Training Set

This document does not specify a separate "training set" sample size in the context of machine learning. For IVD devices, development involves extensive internal testing and optimization (which can be considered analogous to "training"), but the provided document focuses on the validation studies. The studies described are primarily validation studies against regulatory requirements.

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

Given that this is an IVD kit based on turbidimetry, it doesn't involve a "training set" or "ground truth" in the machine learning sense. The "ground truth" during development (analogous to training) would involve using known concentrations of analytes, reference materials, and established analytical methods to ensure the assay performs as expected.

Summary

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August 23, 2018

The Binding Site Group Ltd Andrea Thomas Regulatory Affairs Officer 8 Calthorpe Road Edgbaston Birmingham, B15 1QT Gb

Re: K173732

Trade/Device Name: Optilite Freelite Mx Kappa Free Kit, Optilite Freelite Mx Lambda Free Kit Regulation Number: 21 CFR 866.5550 Regulation Name: Immunoglobulin (light chain specific) immunological test system Regulatory Class: Class II Product Code: DFH, DEH Dated: July 26, 2018 Received: July 31, 2018

Dear Andrea Thomas:

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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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

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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) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; 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 mediation-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.

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For

Lea Carrington, Director Division of Immunology and Hematology 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) K173732

Device Name

Optilite Freelite Mx Kappa Free Kit (LK016.M.OPT.A)

Optilite Freelite Mx Lambda Free Kit (LK018.M.OPT.A)

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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Optilite Freelite Mx Kappa Free Kit and Optilite Freelite Lambda Free Submission Summary

Date prepared: 23d Auqust 2018

A. 510(k) Number:

K173732

B. Purpose for Submission:

Addition of a urine matrix and an addition of a 1+0 sample dilution to a cleared IVD assay (K150658)

C. Measure and:

Kappa (k) free light chains and Lambda (λ) free light chains

D. Type of Test: Quantitative, Turbidimetry

E. Applicant:

The Binding Site Group, Ltd. Contact person: Andrea Thomas Address: 8 Calthorpe Road, Birmingham, B15 1QT, United Kingdom Contact number: +44(0)121 456 9500

F. Proprietary and Established Names:

Optilite® Freelite Mx™ Kappa Free Kit Optilite® Freelite Mx™ Lambda Free Kit

G. Regulatory Information:

1. Requlatory Section: 21 CFR 866.5550, Immunoglobulin (light chain specific) immunological test system
1. Classification: Class II
1. Product Code:

DFH - Kappa antigen, antiserum, control DEH - Lambda antigen, antiserum, control

Panel: Immunology (82)

H. Intended Use:

1. Intended use(s):
  The Optilite Freelite Mx Kappa Free Kit is intended for the quantitative in vitro measurement of Kappa free light chains in serum and urine using the Binding Site Optilite analyser. Measurement of free light chains aids in the diagnosis and monitoring of multiple myeloma, lymphocytic neoplasms, Waldenstrom's macroglobulinaemia, AL amyloidosis, light chain deposition disease and

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connective tissue diseases such as systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings.

Indication(s) for use:

Same as intended use.

1. Special conditions for use statement(s): For prescription use only.
  Warning: The kappa free light chain results for a given specimen determined with assays from different manufacturers or on different systems can vary due to differences in assay methods and reagent specificity. The results reported by the laboratory to the physician must include the identity of the kappa free light chain assay used. Values obtained with different assays or systems cannot be used interchangeably. If, in the course of serially monitoring a patient, the assay or system used for determining kappa free light chain levels is changed, additional sequential testing should be carried out. Prior to changing assay or system, the laboratory MUST confirm baseline values for patients being serially monitored

Warning: The lambda free light chain results for a given specimen determined with assays from different manufacturers or on different systems can vary due to differences in assay methods and reagent specificity. The results reported by the laboratory to the physician must include the identity of the lambda free light chain assay used. Values obtained with different assays or systems cannot be used interchangeably. If, in the course of serially monitoring a patient, the assay or system used for determining lambda free light chain levels is changed, additional sequential testing should be carried out. Prior to changing assay or system, the laboratory MUST confirm baseline values for patients being serially monitored.

Special instrument requirements:

Optilite Analyser (Indiko) (K110035)

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Device Description: .

The Optilite Freelite Mx Kappa Free Kit is comprised of the following reagents:

Latex Reagent: Sheep anti-Human Kappa (polyclonal monospecific) antibody (F(ab')2 fragment) bound to 200nm polystyrene latex particles. In 0.033M Glycine buffered saline pH8 (containing Sodium Azide (0.033%), Proclin (0.05%), Benzamidine (0.01%) and EACA (0.1%) as preservative plus BSA (0.033%)

Calibrator and Controls: Pooled human serum, supplied in stabilised liquid form. Containing 0.099% sodium azide, 0.1% EACA and 0.01% benzamidine as preservatives.

Reaction Buffer: Containing 0.099% sodium azide as a preservative.

The Optilite Freelite Mx Lambda Free Kit is comprised of the following reagents:

Latex Reagent: Sheep anti-Human Lambda (polyclonal monospecific) Free antibody (F(ab')2 fragment) bound to 200nm polystyrene latex particles. In 0.033M Glycine buffered saline pH8 (containing Sodium Azide (0.033%), Proclin (0.05%), Benzamidine (0.01%) and EACA (0.1%) as preservatives plus BSA (0.1665%))

Calibrator and Controls: Pooled human serum, supplied in stabilised liquid form. Containing 0.099% sodium azide, 0.1% EACA and 0.01% benzamidine as preservatives.

Reaction Buffer: Containing 0.099% sodium azide as a preservative.

J. Substantial Equivalence Information:

1. Predicate device names and predicate 510(k) number: Freelite® Human Kappa Free Kit for use on the Siemens BN™II (K) and Freelite® Human Lambda Free Kit for use on the Siemens BN™II (K040009)

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Similarities
Item	Test Device (Optilite)	Predicate (BNII)
Assay type	Quantitative	Same
Intended Use(Kappa)	Quantitation of kappa free light chains in serumand urine. It is intended to aid in the diagnosisand monitoring of multiple myeloma,lymphocytic neoplasms, Waldenström'smacroglobulinaemia, AL amyloidosis, lightchain deposition disease and connective tissuediseases such as systemic lupuserythematosus (SLE) in conjunction with otherlaboratory and clinical findings.	Same
Intended Use(Lambda)	Quantitation of lambda free light chains inserum and urine. It is intended to aid in thediagnosis and monitoring of multiple myeloma,lymphocytic neoplasms, Waldenström'smacroglobulinaemia, AL amyloidosis, lightchain deposition disease and connective tissuediseases such as systemic lupuserythematosus (SLE) in conjunction with otherlaboratory and clinical findings.	Same
Analyte	Kappa Free Light ChainsLambda Free Light Chains	Same
DetectionAntibody	LK016.M.OPT.A: Sheep anti-human kappaantibody (F(ab)2 fragment) bound to latexparticlesLK018.OPT.A: Sheep anti-human kappaantibody (F(ab)2 fragment) bound to latexparticles	Same
Calibration	The Optilite analyser produces a calibrationcurve by performing multiple dilutions of asingle calibrator fluid	Same
Traceability	Internally produced master calibrator	Same
SampleMatrix	Serum and Urine	Same
Open VialStability	3 months	3 months
Differences
Item	Device	Predicate (BNII)
Instrument	Optilite	Siemens BNII Systems
Test Method	Turbidimetric	Nephelometric
AdultReferenceInterval	Free Kappa: 32.90 mg/LFree Lambda: 3.79 mg/L* 97.5th Percentile (one sidedreference interval)	Free Kappa: 1.35 – 24.19 mg/LFree Lambda: 0.24 – 6.66 mg/LKappa/Lambda Ratio: 2.04 - 10.37Reference intervals taken frompredicate product inserts.
MeasuringRange andDilutions(Kappa)	1+0: 0.3 – 12.65 mg/L1+1: 0.6 – 25.3 mg/L1+9: 2.9 – 127 mg/L (Std. dil.)1+99: 29 – 1270 mg/L1+999: 290 – 12700 mg/L1+4999: 1450 – 63500 mg/L	1/1: 0.06 – 1.9 mg/L1/5: 0.3 – 9.5 mg/L1/20: 1.2 – 38.0 mg/L1/100: 5.9 – 190 mg/L (Std. dil.)1/400: 23.6 – 760 mg/L1/2000: 118 – 3800 mg/L1/8000: 472 – 15200 mg/L
MeasuringRange andDilutions(Lambda)	1+0: 0.74 – 17.4 mg/L1+1: 1.3 – 34.7 mg/L1+7: 5.2 – 139 mg/L (Std. dil.)1+79: 52 – 1390 mg/L1+799: 520 – 13900 mg/L1+7999: 5200 – 139000 mg/L	1/1: 0.05 – 1.6 mg/L1/5: 0.25 – 8.0 mg/L1/20: 1.0 – 32.0 mg/L1/100: 5.0 – 160 mg/L (Std. dil.)1/400: 20.0 – 640 mg/L1/2000: 100 – 3200 mg/L1/8000: 400 – 12800 mg/L
On BoardStability	30 days	None quoted

2. Comparison with predicate:

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K. Standard/Guidance Documents Referenced (if applicable):

CLSI EP17-A Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline.

CLSI EP7-A2 Interference Testing in Clinical Chemistry, Approved Guideline -Second Edition.

CLSI EP6-A: Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach.

CLSI EP5-A2 Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline - Second Edition.

CLSI C28-A3c: Defining, Establishing and Verifying Reference Intervals in the Clinical Laboratory.

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L. Test Principle:

Evaluating the concentration of a soluble antigen (e.g. Kappa free light chains) by turbidimetry involves the addition of the test sample to a solution containing the appropriate antibody in a reaction vessel or cuvette. A beam of light is passed through the cuvette and, as the antigen-antibody reaction proceeds, the light passing through the cuvette is scattered increasingly as insoluble immune complexes are formed. Light scatter is monitored by measuring the decrease in intensity of the incident beam of light. The antibody in the cuvette is in excess so the amount of immune complex formed is proportional to the antigen concentration. A series of calibrators of known antigen concentration are assayed initially to produce a calibration curve of measured light scatter versus antigen concentration. Samples of unknown antigen concentration can then be assaved and the results read from the calibration curve.

M. Performance Characteristics (if/when applicable):

1. Analytical performance:
- a. Precision/Reproducibility:

Kappa

The within-run, between-run, between-day and between-instrument precision were determined by testing four urine sample pools over 21 days with two runs per day using one reagent lot on three analysers. Results are summarised below.

Sample	Mean	Within Run		BetweenRun		BetweenDay		BetweenInstrument		Total
		SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
1	5.49	0.28	5.1	0.10	1.8	0.30	5.4	0.35	6.3	0.42	7.6
2	33.08	0.70	2.1	1.61	4.9	2.08	6.3	0.55	1.7	2.72	8.2
3	99.16	3.12	3.1	4.56	4.6	4.29	4.3	2.62	2.7	6.99	7.1
4	179.12	5.16	2.9	5.11	2.9	13.89	7.8	12.88	7.2	15.68	8.8

Lambda

The within-run, between-run, between-day and between-instrument precision were determined by testing five urine sample pools over 23 days with two runs per day using one reagent lot on three analysers. Results are summarised below.

Sample	Mean	Within Run		Between Run		Between Day		Between Instrument		Total
		SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
1	4.48	0.11	2.5	0.22	4.9	0.27	4.8	0.05	1.1	0.33	7.3
2	7.19	0.35	4.8	0.37	5.1	0.36	5.1	0.48	6.7	0.62	8.7
3	54.37	1.22	2.2	3.17	5.8	2.88	5.3	3.25	6.0	4.45	8.2
4	109.27	5.19	4.8	4.39	4.0	5.72	5.2	4.45	4.1	8.89	8.1
5	133.46	4.09	3.1	3.66	2.7	10.11	7.6	11.62	8.7	11.50	8.6

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b. Linearity/assay reportable range:
A linearity study was performed following CLSI document Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach. The linearity of the Kappa Free and Lambda Free assays have been confirmed using serially diluted urine samples to cover the standard measuring ranges of 2.9 – 127mg/L (Kappa) and 5.2 – 139mg/L (Lambda) respectively. The results demonstrated that the kappa free and lambda free assays are linear over the ranges of 2.383-142.687mg/L (Kappa) and 3.87 -160.02mg/L (Lambda) with deviation from linearity ≤ 10%.
c. Traceability, Stability, Expected values (controls, calibrators, or methods): Traceability: Internally produced master calibrator
Stability: Not applicable. On board, open vial and real time stability was evaluated in K150658.
d. Detection limit:
The analytical sensitivity was determined in accordance with CLSI EP17-A and was carried out using both serum and urine matrices. The Limit of Blank was based on 60 determinations of blank samples (analyte depleted serum and urine at a low analyte concentration) and was estimated at the 95th percentile of the distribution. The Limit of Detection was calculated from the LoB and the combined SD of the five LoQ samples. The LoQ was calculated from five independent samples (Serum samples: samples diluted with analyte depleted serum and Urine samples: in house urine at low analyte concentration which was then pooled with an in house urine with a higher analyte concentration) to achieve a concentration close to the bottom of the measuring range) tested twelve times over five days. The tabulated summary of results is shown below:

	LoB	LoD	LoQ
Kappa Free(Serum)	0.195mg/L	0.224mg/L	0.330mg/L
Kappa Free(Urine)	0.210mg/L	0.232mg/L	0.330mg/L
Lambda Free(Serum)	0.285mg/L	0.393mg/L	0.740mg/L
Lambda Free(Urine)	0.080mg/L	0.184mg/L	0.740mg/L

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e. Analytical specificity:

Interference:

Interferences were assessed according to CLSI EP7-A2 by testing a single urine sample for the Kappa Free assay and the Lambda Free assay. Samples were prepared with an analyte concentration around the reference limit of each assay. Each sample was made up using urine from a healthy adult donor and urine from a kappa or lambda myeloma patient. Samples containing interferents were compared to matched control samples which contained no interferent. The mean results from the spiked samples must be within 10% of the mean of the control samples.

Kappa

No significant assay interference effects were observed when the samples were tested with substances at the concentrations given below.

Substance	Concentration
Ascorbic Acid	200 mg/L
Albumin	5 g/L
Bilirubin	100 mg/L
Bortezomib	6 mg/mL
Cyclophosphamide	330 mg/mL
Digoxin	0.375 $ \mu $ g/mL
Haemoglobin	240 mg/L
Penicillin	75 mg/L
Phenytoin	15 mg/L
Theophylline	150 mg/L

Lambda

No significant assay interference effects were observed when the samples were tested with substances at the concentrations given below.

Substance	Conc.
Ascorbic Acid	200 mg/L
Albumin	5 g/L
Bilirubin	100 mg/L
Bortezomib	3.6 mg/mL
Cyclophosphamide	330 mg/mL
Digoxin	0.375 µg/mL
Haemoglobin	250 mg/L
Penicillin	75 mg/L
Phenytoin	15 mg/L
Theophylline	150 mg/L

Cross reactivity:

No significant cross reaction was observed during testing for the predicate device. The specificity of the antisera is unchanged.

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Antigen Excess Detection:

The possibility of antigen excess occurring when using the device on the Optilite analyser was evaluated in K150658

Assay cut-off:

Not applicable. Refer to expected values of reference range.

1. Comparison studies:
- a. Method comparison with predicate device:

Kappa Free:

A comparison study was performed by analysing 165 urine samples (including 86 with analyte levels below the reference limit) using the Optilite Freelite Mx Kappa Free kit and an alternative commercially available assay. Passing Bablok regression analysis generated the following results:

y = 1.02x - 0.21 (mg/L) (y = Optilite; x = predicate analyser) correlation coefficient r = 0.992

Lambda Free:

A comparison study was performed by analysing 115 urine samples (including 59 with analyte levels below the reference limit) using the Optilite Freelite Mx Lambda Free kit and an alternative commercially available assay. Passing Bablok regression analysis generated the following results:

y = 1.11x — 0.27 (mg/L) (y = Optilite; x = predicate analyser) correlation coefficient r = 0.947

b. Matrix comparison: Not applicable.

3. Clinical studies:

a. Clinical Sensitivity/clinical specificity:
Kappa (2X2):

		Predicate Assay
		+	-	Total
Test Assay	+	78	1	79
		-	8	79
	Total	86	80	166

Agreement	Percentage	95% ConfidenceInterval
Positive	90.7%	82.7% to 95.2%
Negative	98.8%	93.3% to 99.8%
Overall	94.6%	90.0% to 97.1%

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Lambda (2X2):

		Predicate Assay
		+	-	Total
Test Assay	+	56	0	56
	-	20	92	112
	Total	76	92	168

Agreement	Percentage	95% ConfidenceInterval
Positive	73.7%	62.8% to 82.3%
Negative	100.0%	96.0% to 100.0%
Overall	88.1%	92.3% to 92.2%

b. Other clinical supportive data (when a. is not applicable): Not applicable.
1. Clinical cut-off: Not applicable
1. Expected values/Reference range:

Normal adult urine	Mean(mg/L)	Median(mg/L)	Reference Limit(mg/L)(97.5th Percentile)
Kappa free	7.64	4.54	32.90
Lambda free	1.03	0.74	3.79

N. Proposed Labelling:

The labelling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

Regulation Number and Section

§ 866.5550 Immunoglobulin (light chain specific) immunological test system.

(a)
Identification. An immunoglobulin (light chain specific) immunological test system is a device that consists of the reagents used to measure by immunochemical techniques both kappa and lambda types of light chain portions of immunoglobulin molecules in serum, other body fluids, and tissues. In some disease states, an excess of light chains are produced by the antibody-forming cells. These free light chains, unassociated with gamma globulin molecules, can be found in a patient's body fluids and tissues. Measurement of the various amounts of the different types of light chains aids in the diagnosis of multiple myeloma (cancer of antibody-forming cells), lymphocytic neoplasms (cancer of lymphoid tissue), Waldenstrom's macroglobulinemia (increased production of large immunoglobulins), and connective tissue diseases such as rheumatoid arthritis or systemic lupus erythematosus.(b)
Classification. Class II (performance standards).