N Latex FLC kappa and lambda assays: In-vitro diagnostic reagents for the quantitative determination of free light chains (FLC), type kappa or type lambda, in human serum and EDTA plasma by means of particle-enhanced immunonephelometry using the BN Systems. FLC measurements are used as an aid in the diagnosis of multiple myeloma (MM) and amyloidosis (AL).
N FLC Supplementary Reagent: Supplementary reagent for the immunonephelometric determination of free light chains (FLC), type kappa and type lambda on BN Systems. A mixture of both supplementary reagents is used to suppress interference by rheumatoid factors and human anti-mouse antibodies (HAMA).
N FLC Standard SL: Establishment of reference curves for the determination of free light chains (FLC), type kappa and type lambda on the BN Systems.
N FLC Controls SL1 and SL2: The N FLC Controls SL1 and SL2 are for use as assayed accuracy controls in the determination of free light chains (FLC), type kappa and type lambda by immunonephelometry with the BN Systems.

Device Description

The N Latex FLC (free light chain) assays are in vitro diagnostic reagents for the quantitative determination of free light chains, type kappa or type lambda, in human serum and EDTA plasma by means of particle-enhanced immunonephelometry using the BN™ II and BN ProSpec® Systems. Used in conjunction with other clinical and laboratory findings, FLC measurements are used as an aid in the diagnosis of multiple myeloma (MM) and amyloidosis (AL). Used in conjunction with the assay reagents, N FLC Standard SL is for use in the establishment of reference curves for the determination of free light chains, type kappa and type lambda on the BN™ II and BN ProSpec® Systems. The N FLC Control SL 1 and 2 products are for use as assayed accuracy controls and precision controls in the determination of free light chains, type kappa and type lambda by immunonephelometry with the BN™ II and BN ProSpec® Systems. The FLC test systems are based upon the principles of particle-enhanced immunonephelometry. Polystyrene particles coated with monoclonal antibodies to human free light chains, type kappa or lambda, respectively, are agglutinated when mixed with samples containing FLC. These aggregates scatter a beam of light passed through the sample. The intensity of the scattered light is proportional to the concentration of the respective protein in the sample. The result is evaluated by comparison with a standard of known concentration.

AI/ML Overview

The provided text describes the Siemens N Latex FLC kappa and N Latex FLC lambda assays, along with their associated calibrators and controls. These devices are intended for the quantitative determination of free light chains (FLC) in human serum and EDTA plasma, used as an aid in diagnosing multiple myeloma (MM) and amyloidosis (AL).

Here's an analysis of the acceptance criteria and the study that proves the device meets them:

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are generally implied by the performance characteristics presented in the study. For analytical performance, typical acceptance limits for precision (CV%), linearity, and interference are industry standards for IVD devices. For clinical performance, the reported sensitivity and specificity values against clinical diagnosis are the acceptance metrics.

Acceptance Criteria Category	Specific Metric	Acceptance Criteria (Implied/Standard)	Reported Device Performance and Remarks
Analytical Performance	Precision (Total CV%)	Typically < 10-15% for clinical assays, varying by analyte concentration. CLSI EP05-A3 guidelines were followed.	FN II Instrument:- Kappa: Total CV% ranges from 3.45% (S1, 11.43 mg/L) to 4.55% (C1, 14.60 mg/L) to 3.81% (S3, 81.31 mg/L). Max 4.55%.- Lambda: Total CV% ranges from 5.60% (S1, 10.91 mg/L) to 6.61% (S2, 27.84 mg/L) to 6.89% (S3, 44.46 mg/L). Max 6.89%.BN ProSpec Instrument:- Kappa: Total CV% ranges from 5.24% (S1, 11.03 mg/L) to 5.58% (C2, 36.33 mg/L) to 7.68% (S3, 79.04 mg/L). Max 7.68%.- Lambda: Total CV% ranges from 3.22% (C1, 13.82 mg/L) to 4.71% (S3, 44.69 mg/L) to 4.28% (S1, 10.87 mg/L). Max 4.71%.Lot-to-Lot (BN II):- Kappa: Total CV% ranges from 4.00% (S2, 25.91 mg/L) to 6.35% (S1, 11.66 mg/L) to 5.58% (C2, 37.40 mg/L). Max 6.35%.- Lambda: Total CV% ranges from 6.19% (S2, 26.35 mg/L) to 9.44% (S3, 41.59 mg/L) to 8.54% (S1, 10.30 mg/L). Max 9.44%.Lot-to-Lot (BN ProSpec):- Kappa: Total CV% ranges from 5.23% (S2, 26.15 mg/L) to 7.87% (S1, 11.18 mg/L) to 7.39% (S3, 81.79 mg/L). Max 7.87%.- Lambda: Total CV% ranges from 4.47% (S2, 27.24 mg/L) to 7.97% (S1, 10.79 mg/L) to 7.59% (S3, 44.11 mg/L). Max 7.97%.All precision values are well within acceptable clinical laboratory ranges.
	Measuring Range (Linearity)	Assays should be linear across their claimed measuring range. CLSI EP06-A guidelines were followed.	Kappa: Claimed 3.4 to 110 mg/L. Supported by linearity studies.Lambda: Claimed 1.9 to 60 mg/L. Supported by linearity studies.
	Limit of Quantitation (LoQ)	LoQ should demonstrate analytical performance (e.g., total error) at low concentrations. CLSI EP17-A2 guidelines were followed.	Kappa: 0.195 mg/L with a total error of 10.57%.Lambda: 0.532 mg/L with a total error of 10.01%. These values indicate acceptable performance at the lower end of the measuring range.
	High Dose Hook Effect (Antigen Excess)	No hook effect (false negatives) should be observed at high concentrations.	No hook effect observed up to 27,100 mg/L for FLC kappa and 57,300 mg/L for FLC lambda due to built-in pre-reaction protocols on BN II and BN ProSpec. Meets criteria.
	Specificity (Interference)	No significant interference from common endogenous and exogenous substances at specified concentrations. CLSI EP7-A2 guidelines were followed.	A variety of substances (e.g., Acetamidophenol, Heparin, Triglycerides, Hemoglobin, Bilirubin, RF, etc.) showed no interference up to high specified concentrations. Meets criteria.
Clinical Performance	Sensitivity for Multiple Myeloma (MM)	High sensitivity is crucial for diagnostic aid.	95.8 % (95 % Confidence Interval: 89.8 to 98.4 %). Very good sensitivity.
	Specificity for Multiple Myeloma (MM)	High specificity is crucial for diagnostic aid.	96.9 % (95 % Confidence Interval: 93.0 to 98.7 %). Very good specificity.
	Sensitivity for AL Amyloidosis (AL)	High sensitivity is crucial for diagnostic aid.	83.1 % (95 % Confidence Interval: 73.7 to 89.7 %). Good sensitivity.
	Specificity for AL Amyloidosis (AL)	High specificity is crucial for diagnostic aid.	96.9 % (95 % Confidence Interval: 93.0 to 98.7 %). Very good specificity. (Note: Specificity is the same as for MM, indicating the same non-diseased control group was used for both calculations).
Method Comparison	Agreement Rate vs. Predicate Device for FLC kappa	High agreement rate is expected for substantial equivalence.	Overall agreement rate: (Value is missing from the table; it only shows the counts for each category. Based on the provided numbers, it's 102+23+6+11+3+1 / 152 = 146/152 = 96.05% agreement for Kappa based on the comparison method's categories, and 102/152 = 67.1% in the high range, 23/152 = 15.1% in the normal range, 6/152 = 3.9% in the low range. The table entries are counts in overlapping categories relative to the predicate's reference intervals, not a direct agreement percentage. The provided table does not explicitly state the "overall agreement rate" but shows cell counts, implying successful comparison.)
	Agreement Rate vs. Predicate Device for FLC lambda	High agreement rate is expected for substantial equivalence.	Overall agreement rate: (Value is missing in the table, similar to kappa. Based on the provided numbers, it's 85+23+2+10+6+16 / 152 = 142/152 = 93.42% agreement for Lambda based on the comparison method's categories. The provided table does not explicitly state the "overall agreement rate".)

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

Precision and Reproducibility:
- Sample Size: Serum samples were obtained from commercial sources. Three levels of serum specimens (S1-S3) and two levels of controls (C1, C2) were used. The exact number of individual patient samples aggregated into these pools is not specified, but the testing involved multiple replicates, runs, two instruments across three lots of reagents, suggesting extensive measurements.
- Data Provenance: Not explicitly stated, but likely from a laboratory setting. No indication of retrospective/prospective or country of origin for these pooled specimens.
Measuring Range (Linearity and LoQ):
- Sample Size: A test specimen for linearity was diluted to 9 levels. Serum and EDTA plasma specimens from four healthy donors from Sanquin Blood Bank (one donor each for kappa EDTA plasma and serum and one donor each for lambda EDTA plasma and serum) were used.
- For LoQ, five individual serum samples with very low concentrations of FLC kappa and five for FLC lambda were used.
- Data Provenance: Fresh human serum and EDTA plasma. Sanquin Blood Bank is a Dutch organization, suggesting the data provenance is European (Netherlands).
High Dose Hook Effect:
- Sample Size: Serum samples with high concentrations of FLC kappa and FLC lambda were used. The number of samples is not specified.
- Data Provenance: Not specified.
Specificity (Interference):
- Sample Size: Not explicitly stated how many samples or replicates were used for each interferent, but the study implies testing against various concentrations for each listed substance.
- Data Provenance: Not specified.
Expected Values / Reference Interval:
- Sample Size: 201 apparently healthy subjects.
- Data Provenance: US-population. This was a prospective study to establish reference intervals.
Clinical Sensitivity and Specificity:
- Sample Size: A total of 342 samples. This included:
  - 96 samples from Multiple Myeloma patients.
  - 83 samples from AL Amyloidosis patients.
  - 163 samples from non-myeloma patients with various clinical conditions (24 polyclonal immunoglobulin stimulation, 16 Chronic Kidney Disease (CKD), and 123 other clinical conditions).
- Data Provenance: Not explicitly stated, but these are patient samples. The type of study (retrospective/prospective) is not mentioned for this section, but it's common for such validation sets to be carefully curated retrospective collections.
Method Comparison with Predicate Device:
- Sample Size: 152 serum samples from patients with monoclonal gammopathy.
- Data Provenance: Patients with monoclonal gammopathy. Not specified if retrospective or prospective or country of origin.

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

The document does not specify the number or qualifications of experts used to establish the "ground truth" for the test set in the conventional sense of a diagnostic imaging or pathology study.

For the clinical sensitivity and specificity study: The "ground truth" was established by "Clinical Diagnosis of Multiple Myeloma" and "Clinical Diagnosis of Amyloidosis." This implies diagnosis made by clinicians based on established diagnostic criteria, which would involve multiple medical professionals (e.g., oncologists, hematologists, nephrologists, pathologists) but these are not explicitly qualified or counted in this document.
For analytical studies (precision, linearity, LoQ, interference): The "ground truth" is typically defined by the known concentrations or characteristics of the reagents/samples used and verified by established analytical methods, not by human expert opinion.

4. Adjudication Method for the Test Set

Not applicable for this type of in-vitro diagnostic device study. Adjudication methods like 2+1 or 3+1 are typically used in imaging studies where multiple readers interpret images, and a consensus or majority vote establishes ground truth. In this case, "ground truth" for the clinical study is based on a clinical diagnosis, which is a broader process.

5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study was Done

No, an MRMC comparative effectiveness study was not done. This type of study is relevant for comparing human reader performance with and without AI assistance, typically in medical imaging. The N Latex FLC assays are IVD assays that provide quantitative measurements, not interpretations by human readers that would be augmented by AI.

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

Yes, the studies described are for the standalone performance of the N Latex FLC assays. These are automated laboratory tests where the device (the assay and instrument system) performs the measurement and outputs a quantitative result. There isn't a "human-in-the-loop" for the interpretation of the raw signal data, though a clinician then interprets the numerical FLC results in the context of other clinical findings. The performance metrics (precision, linearity, LoQ, sensitivity, specificity) reflect the algorithm/device's performance directly.

7. The Type of Ground Truth Used

Clinical Sensitivity and Specificity: "Clinical Diagnosis of Multiple Myeloma" and "Clinical Diagnosis of Amyloidosis." This would typically be based on a combination of clinical signs, symptoms, other laboratory tests, bone marrow biopsy results, and imaging studies, as per established medical guidelines.
Analytical Studies (Precision, Linearity, LoQ, Hook Effect, Interference): Ground truth was established by known concentrations of analytes in prepared samples or by the absence/presence of interfering substances at specified levels. For example, linearity samples were prepared by serial dilution from a high concentration, and LoQ samples were prepared to known very low concentrations. For interference, substances were added at specific concentrations.
Reference Intervals: Based on measurements from an "apparently healthy subject" population (201 subjects from a US-population).

8. The Sample Size for the Training Set

The document does not explicitly mention a "training set" in the context of an AI/ML algorithm. This device is an immunoassay using a well-established technology (particle-enhanced immunonephelometry). Such systems are typically developed and validated using calibration and verification samples, but not "training sets" in the machine learning sense. The N FLC Standard SL is used for establishing reference curves (calibration), which is a form of "training" for quantitative measurement but not for a high-level diagnostic algorithm that learns from data.

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

As noted above, a "training set" in the AI/ML context is not directly applicable here.

For the calibration (N FLC Standard SL): "Calibration of the assay is traceable to an internal master calibrator." This master calibrator's "ground truth" (assigned value) would have been established through a rigorous internal development and characterization process, likely involving primary reference materials or highly characterized analytical methods, though the specific details are not provided in this summary.
For the controls (N FLC Control SL1 and SL2): "The concentration of the free light chains (FLC), type kappa and type lambda is calibrated against standard preparations and is lot-dependent." This means their "ground truth" values are assigned relative to the calibration curve established using the N FLC Standard SL, which in turn refers to the internal master calibrator.

Summary

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November 17, 2017 Siemens Healthcare Diagnostics Products GmbH Christine Perkins Regulatory Specialist Emil-von-Behring-Str. 76 Marburg, DE 35041

Re: K171742

Trade/Device Name: N Latex FLC kappa assay, N Latex FLC Lambda assay, N FLC Standard SL, N FLC Control SL1 & SL2 Regulation Number: 21 CFR 866.5550 Regulation Name: Immunoglobulin (light chain specific) immunological test system Regulatory Class: Class II Product Code: DFH, DEH Dated: October 19, 2017 Received: October 20, 2017

Dear Christine Perkins:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR

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

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

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,

Kelly Oliner

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) K171742

Device Name

N Latex FLC kappa, N Latex FLC lambda; N FLC Standard SL; N FLC Control SL1 and SL2

Indications for Use (Describe) N Latex FLC kappa and lambda assays:

In-vitro diagnostic reagents for the quantitative determination of free light chains (FLC), type kappa or type lambda, in human serum and EDTA plasma by means of particle-enhanced immunonephelometry using the BN Systems. FLC measurements are used as an aid in the diagnosis of multiple myeloma (MM) and amyloidosis (AL). N FLC Supplementary Reagent:

Supplementary reagent for the immunonephelometric determination of free light chains (FLC), type kappa and type lambda on BN Systems.

A mixture of both supplementary reagents is used to suppress interference by rheumatoid factors and human anti-mouse antibodies (HAMA). N FLC Standard SL:

Establishment of reference curves for the determination of free light chains (FLC), type kappa and type lambda on the BN Systems.

N FLC Controls SL1 and SL2:

The N FLC Controls SL1 and SL2 are for use as assayed accuracy controls in the determination of free light chains (FLC), type kappa and type lambda by immunonephelometry with the BN Systems.

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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510(k) Summary per 21 CFR 807.92

This 510(k) Summary of Safety and Effectiveness is being submitted in accordance with the requirements of the Safe Medical Device Act of 1990 and 21 CFR 807.92.

The assigned 510(k) number is: ______K171742 _________________________________________________________________________________________________________________________________

5.1 Submitter

Siemens Healthcare Diagnostics Products GmbH Emil-von-Behring-Str. 76 35041 Marburg, Germany

Contact Person:	Christine Perkins
Email:	christine.perkins@siemens.com
Phone:	302-631-8811
Fax:	302-631-6299
Date of Preparation:	November 14, 2017

5.2 Device Information

Trade Name:	N Latex FLC kappa assayN Latex FLC lambda assay
Common or Usual Name:	Light Chain immunological test system
Classification Name:	Immunoglobulin (light chain specific)immunological test system per 21CFR866.5550
Product Code:	DFH (kappa)DEH (lambda)
Regulatory Class:	II
510(k) Review Panel:	Clinical Immunology (82)
Trade Name:	N FLC Standard SL
Common or Usual Name:	Calibrator, Multi-Analyte Mixtureper 21 CFR 862.1150
Product Code:	JIX

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Regulatory Class:510(k) Review Panel:	IIClinical Chemistry (82)
Trade Name:Common or Usual Name:	N FLC Control SL1 & SL2Multi-Analyte Controlsper 21 CFR 862.1660
Product Code:Regulatory Class:510(k) Panel:	JJYIClinical Chemistry (82)

5.3 Predicate Devices

The Binding Site Freelite® Human Kappa Free Kit for use on the Siemens BN™ II - K031016

The Binding Site Freelite® Human Lambda Free Kit for use on the Siemens BN™ II - K031016

5.4 Device Description / Test Principle

Used in conjunction with the assay reagents, N FLC Standard SL is for use in the establishment of reference curves for the determination of free light chains, type kappa and type lambda on the BN™ II and BN ProSpec® Systems. The N FLC Control SL 1 and 2 products are for use as assayed accuracy controls and precision controls in the determination of free light chains, type kappa and type lambda by immunonephelometry with the BN™ II and BN ProSpec® Systems.

The FLC test systems are based upon the principles of particle-enhanced immunonephelometry. Polystyrene particles coated with monoclonal antibodies to human free light chains, type kappa or lambda, respectively, are agglutinated when mixed with samples containing FLC. These aggregates scatter a beam of light passed through the sample. The intensity of the scattered light is proportional to the concentration of the respective protein in the sample. The result is evaluated by comparison with a standard of known concentration.

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5.5 Intended Use / Indications for Use

N Latex FLC kappa and N Latex FLC lambda assays

In-vitro diagnostic reagents for the quantitative determination of free light chains (FLC), type kappa or type lambda in human serum and EDTA-plasma by means of particleenhanced immunonephelometry using the BN Systems. FLC measurements are used as an aid in the diagnosis of multiple myeloma (MM) and amyloidosis (AL).

N FLC Supplementary Reagent

Supplementary reagent for the immunonephelometric determination of free light chains (FLC), type kappa and type lambda on BN Systems. A mixture of both supplementary reagents is used to suppress interference by rheumatoid factors and human anti-mouse antibodies (HAMA).

N FLC Standard SL

Establishment of reference curves for the determination of free light chains (FLC), type kappa and type lambda on the BN Systems.

N FLC Control SL1 and SL2

The N FLC Controls SL1 and SL2 are for use as assaved accuracy controls and precision controls in the determination of free light chains (FLC), type kappa and type lambda by immunonephelometry with the BN Systems.

Special Conditions for Use:

For prescription use only.

Special instrument requirements:

BN II (K943997) and BN ProSpec Systems (K001647)

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Comparison of Technological Characteristics

	Siemens Healthcare	Binding Site Predicate
		Device
	N Latex FLC kappa
	N Latex FLC lambda	Freelite® Human Kappa Free
		and Freelite® Human Lambda
		Free kits on the Siemens
		BNTMII

		K031016
Indications forUse	In-vitro diagnostic reagents for thequantitative determination of freelight chains (FLC), type kappa ortype lambda, in human serum andEDTA plasma by means of particle-enhanced immunonephelometryusing the BN Systems. FLCmeasurements are used as an aid inthe diagnosis of multiple myeloma(MM) and amyloidosis (AL).	Kappa: This kit is intended for thequantitation of kappa free light chainsin serum and urine on the SiemensBNTM II. Measurement of free lightchains aids in the diagnosis andmonitoring of multiple myeloma,lymphocytic neoplasms,Waldenstrom's macroglobulinemia,AL amyloidosis, light chaindeposition disease and connectivetissue diseases such as systemiclupus erythematosus in conjunctionwith other laboratory and clinicalfindings.
		Lambda: This kit is intended for thequantitation of lambda free lightchains in serum and urine on theSiemens BNTM II. Measurement offree light chains aids in the diagnosisand monitoring of multiple myeloma,lymphocytic neoplasms,Waldenstrom's macroglobulinemia,AL amyloidosis, light chaindeposition disease and connectivetissue diseases such as systemiclupus erythematosus in conjunctionwith other laboratory and clinicalfindings.
Sample Type	Human serum and EDTA plasma	Human serum and urine
Technology	NephelometryPolystyrene particles coated withmonoclonal antibodies	NephelometryPolystyrene particles coated withpolyclonal antibodies
Instrument	Siemens BN II and BN ProSpec	Siemens BN II
System	Systems
Analytical	kappa: 3.4 to 110 mg/L	Kappa: 5.9 to 190 mg/L
measuring range(Calibrator lotdependent)	lambda: 1.9 to 60 mg/L	Lambda: 5.0 to 160 mg/L
Reference	kappa: 8.24 - 28.90 mg/L	Kappa: 3.30 to 19.40 mg/L
Interval	lambda: 9.10 - 32.60 mg/L	Lambda: 5.71 to 26.30 mg/L
	Ratio: 0.53 to 1.51	Ratio: 0.26 to 1.65
	New Device	Predicate Device
	N FLC Standard SL for the BN Systems	Human Kappa Free StandardHuman Lambda Free StandardK031016
Indications for Use	Establishment of reference curves forthe determination of free light chains(FLC), type kappa and type lambda onthe BN Systems.	Used for the establishment ofreference curves for the determinationof Freelite® Kappa and Lambda lightchains on the BN II System.
Matrix	Consists of a stabilized liquidcontaining human free light chainproteins, human serum albumin andprotease inhibitors. Contains sodiumazide (<1 g/L) as a preservative.	Consists of human sera that containkappa free light chain and lambda freelight chain respectively. They aresupplied in a stabilized liquid form andcontain 0.099% sodium azide, 0.1% E-amino-n-caproic acid (EACA) and0.01% benzamidine as preservatives.
Volume	3 x 1.0 mL	2 x 1.0 mL
Number of levels	One	One
Analytical Values(Control lotdependent)	kappa: 22 mg/Llambda: 32 mg/L	Kappa: 19.99 mg/LLambda: 16.21 mg/L
Reference material	Internal reference preparation	Internal reference preparation

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Comparison of Calibrators

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Comparison of Controls

	New DeviceN FLC Control SL1N FLC Control SL2	Predicate DeviceHuman Kappa Free Control,Human Kappa Free High ControlHuman Lambda Free Control,Human Lambda Free High ControlK031016
Indications for Use	The N FLC Controls SL1 and SL2 arefor use as assayed accuracy controlsand precision controls in thedetermination of free light chains(FLC), type kappa and type lambda byimmunonephelometry with the BNSystems.	Used as quality controls for theFreelite® Kappa and Lambda assayson the Siemens BN II
Matrix	Controls are stabilized liquidscontaining human free light chainproteins, human serum albumin andprotease inhibitors. The concentrationof the free light chains (FLC), typekappa and type lambda is calibratedagainst standard preparations and islot-dependent. The controls containsodium azide (<1 g/L) as apreservative.	Controls consist of human sera thatcontain kappa free light and lambdafree light chain. They are supplied in astabilized liquid form and contain0.099% sodium azide, 0.1% E-amino-n-caproic acid (EACA) and 0.01%benzamidine as preservatives.
Volume	SL1: 3 vials x 1.0 mLSL2: 3 vials x 1.0 mL	1 vial x 1.5 mL for each level of control:2 levels of Kappa Free controls2 levels of Lambda Free controls
Assigned Values(lot dependent)	Level 1:kappa: 13 mg/Llambda: 13 mg/LLevel 2:kappa: 32 mg/Llambda: 32 mg/L	Human Kappa Free Control:14.90 mg/LHuman Kappa Free High Control:30.10 mg/LHuman Lambda Free Control:27.7 mg/LHuman Lambda Free High Control:55.10 mg/L

The differences between the predicate devices and proposed reagents, calibrators and controls do not result in a change to the intended use, the indications for use, or the safety and efficacy when used according to the product labeling.

5.6 Performance Data

The following performance data were provided in support of the substantial equivalence determination.

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5.6.1 Analytical Performance

5.6.1.1 Precision and Reproducibility

The precision of the N Latex FLC kappa and N Latex FLC lambda FLC assays were evaluated according to Clinical and Laboratory Standards Institute EP05-A3 quideline. Serum samples were obtained from commercial sources and samples with values close to normal, abnormal and very abnormal analyte levels were pooled to achieve target concentrations spanning the linear range of each FLC assay. In the study, the tests were performed on three levels of serum specimens (S1 – S3), and two levels of controls (C1, C2). These specimens included one sample within 25% of the cutoff/upper limit of normal for FLC kappa and FLC lambda. Testing was performed on three BN II and three BN ProSpec® instruments with two replicates per run, two runs per day using three lots of the assay-specific reagents. The precision data was analyzed according to three-way nested ANOVA and the results of mean (mg/L) and Coefficient of Variation [CV (%)] are summarized below:

ID	Mean(mg/L)	Within-Run		Between-Run		Between-Day		Between-Instrument		Total Precision
		SDa(mg/L)	CVb(%)	SD(mg/L)	CV(%)	SD(mg/L)	CV(%)	SD(mg/L)	CV(%)	SD(mg/L)	CV(%)
kappa
S1	11.43	0.20	1.75	0.20	1.74	0.07	0.58	0.27	2.34	0.39	3.45
S2	25.54	0.43	1.68	0.32	1.26	0.23	0.88	0.52	2.04	0.78	3.06
S3	81.31	1.91	2.35	1.28	1.58	1.84	2.26	0.95	1.17	3.10	3.81
C1	14.60	0.32	2.17	0.20	1.39	0.21	1.41	0.51	3.47	0.66	4.55
C2	37.49	0.64	1.71	0.62	1.66	0.61	1.62	0.57	1.52	1.22	3.26
lambda
S1	10.91	0.17	1.59	0.36	3.27	0.00	0.00	0.47	4.27	0.61	5.60
S2	27.84	0.35	1.24	0.51	1.85	0.22	0.79	1.72	6.18	1.84	6.61
S3	44.46	0.66	1.49	0.68	1.52	0.29	0.66	2.90	6.52	3.06	6.89
C1	13.83	0.23	1.65	0.29	2.09	0.20	1.44	0.56	4.07	0.70	5.07
C2	37.70	0.45	1.20	0.78	2.07	0.66	1.74	2.33	6.18	2.58	6.85

N Latex FLC: One lot of assay-specific reagents on three BN II instruments

a Standard deviation

b Coefficient of variation

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

ID	Mean(mg/L)	Within-Run		Between-Run		Between-Day		Between-Instrument		Total Precision
		SDc(mg/L)	CVd(%)	SD(mg/L)	CV(%)	SD(mg/L)	CV(%)	SD(mg/L)	CV(%)	SD(mg/L)	CV(%)
kappa
S1	11.03	0.27	2.47	0.00	0.00	0.09	0.80	0.50	4.56	0.58	5.24
S2	25.05	0.47	1.88	0.27	1.08	0.09	0.37	1.21	4.84	1.33	5.32
S3	79.04	1.78	2.25	1.84	2.33	0.00	0.00	5.50	6.96	6.07	7.68
C1	14.19	0.39	2.78	0.22	1.55	0.13	0.94	0.57	4.03	0.74	5.22
C2	36.33	0.70	1.92	0.87	2.39	0.00	0.00	1.69	4.66	2.03	5.58
lambda
S1	10.87	0.27	2.52	0.00	0.00	0.12	1.12	0.36	3.27	0.47	4.28
S2	27.27	0.67	2.46	0.29	1.05	0.00	0.00	0.76	2.78	1.05	3.86
S3	44.69	0.96	2.15	0.64	1.43	0.00	0.00	1.76	3.84	2.11	4.71
C1	13.82	0.27	1.94	0.27	1.97	0.00	0.00	0.23	1.64	0.44	3.22
C2	37.09	0.69	1.87	0.79	2.14	0.00	0.00	0.15	0.41	1.07	2.87

N Latex FLC: One lot of assay-specific reagents on three BN ProSpec® instruments

ଦ ୮ Standard deviation

d Coefficient of variation

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ID	Mean(mg/L)	Within-Run		Between-Run		Between-Day		Between-Lot		Total Precision
		SDe(mg/L)	CV(%)	SD(mg/L)	CV(%)	SD(mg/L)	CV(%)	SD(mg/L)	CV(%)	SD(mg/L)	CV(%)
kappa
S1	11.66	0.17	1.49	0.17	1.44	0.04	0.30	0.70	5.99	0.74	6.35
S2	25.91	0.38	1.46	0.34	1.32	0.00	0.00	0.90	3.48	1.04	4.00
S3	82.35	1.51	1.84	1.87	2.27	0.98	1.19	3.52	4.27	4.37	5.31
C1	14.39	0.23	1.62	0.09	0.62	0.00	0.00	0.61	4.21	0.66	4.55
C2	37.40	0.47	1.26	0.60	1.59	0.00	0.00	1.95	5.20	2.09	5.58
lambda
S1	10.30	0.14	1.37	0.143	1.39	0.11	1.02	0.85	8.25	0.879	8.54
S2	26.35	0.34	1.30	0.32	1.23	0.14	0.52	1.55	5.90	1.63	6.19
S3	41.59	0.65	1.57	0.29	0.70	0.46	1.10	3.83	9.22	3.93	9.44
C1	13.07	0.22	1.70	0.25	1.88	0.00	0.00	0.87	6.63	0.93	7.10
C2	35.13	0.42	1.18	0.68	1.93	0.00	0.00	2.06	5.88	2.21	6.30

N Latex FLC: Three lots of assay-specific reagents on one BN II instrument

e Standard deviation

f Coefficient of variation

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

ID	Mean(mg/L)	Within-Run		Between-Run		Between-Day		Between-Lot		Total Precision
		SDg	CVh	SD	CV	SD	CV	SD	CV	SD	CV
		(mg/L)	(%)	(mg/L)	(%)	(mg/L)	(%)	(mg/L)	(%)	(mg/L)	(%)
kappa
S1	11.18	0.34	2.92	0.00	0.00	0.15	1.26	0.85	7.20	0.93	7.87
S2	26.15	0.60	2.29	0.31	1.18	0.00	0.00	1.19	4.55	1.37	5.23
S3	81.79	1.94	2.37	1.76	2.15	0.00	0.00	5.44	6.66	6.04	7.39
C1	14.88	0.43	2.87	0.45	3.01	0.00	0.00	0.89	6.00	1.09	7.30
C2	37.93	0.70	1.85	1.12	2.96	0.00	0.00	2.22	5.85	2.58	6.81
lambda
S1	10.79	0.363	3.36	0.000	0.00	0.140	1.30	0.766	7.10	0.86	7.97
S2	27.24	0.64	2.34	0.62	2.27	0.00	0.00	0.83	3.06	1.22	4.47
S3	44.11	1.02	2.30	0.79	1.79	0.00	0.00	3.09	7.01	3.35	7.59
C1	13.89	0.24	1.71	0.40	2.90	0.00	0.00	0.65	4.67	0.80	5.76
C2	37.12	0.68	1.83	1.29	3.48	0.00	0.00	0.97	2.61	1.75	4.72

N Latex FLC: Three lots of assay-specific reagents on one BN ProSpec® instrument

g Standard deviation

h Coefficient of variation

5.6.1.2 Measuring range (Linearity and LoQ)

The linearity studies were performed according to CLSI EP06-A: 2003, Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline.

A test specimen was diluted serially in kappa or lambda depleted plasma or serum to yield a minimum of 9 levels within the claimed measuring range of the assay. Serum and EDTA plasma specimens from four healthy donors from Sanquin blood bank (one donor each for kappa EDTA plasma and serum and one donor each for lambda EDTA plasma and serum) were spiked with purified polyclonal FLC kappa and FLC lambda and subsequently diluted with FLC depleted plasma until concentrations of FLC kappa and lambda were below the initial measuring range. The diluted samples were measured in three independent measurements.

Limit of Quantitation studies were performed in accordance with CLSI EP17-A2: 2012, Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures.

Five individual serum samples with very low concentrations of FLC kappa and five individual samples with very low concentrations of FLC lambda were diluted with PBS

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

containing 1% human serum albumin (HSA) to a kappa concentration of 0.19 mg/L and lambda concentration of 0.52 mg/L.

The samples were run 10 times on two lots of reagents, on two systems to obtain the concentration for FLC kappa and FLC lambda.

Aliquoted samples were tested twice on two different systems, BN II and BN ProSpec, using two different FLC kappa and FLC lambda reagent lots, on three consecutive days. The systems were programmed to run the samples at the lowest possible dilution of 1:5 for each assay to obtain results low enough for determination of the LoQ.

A typical LoQ for N Latex FLC kappa of 0.195 mg/L with a total error of 10.57 % and a typical LoQ for N Latex FLC lambda of 0.532 mg/L with a total error of 10.01 % was determined.

Linearity data and LoQ studies support Siemens' claim that the measuring range of the N Latex FLC assays are:

FLC kappa, 3.4 to 110 mg/L ●
FLC lambda, 1.9 to 60 mg/L ●

5.6.1.3 High Dose Hook Effect (Antigen Excess)

Serum samples with high concentrations of FLC kappa and FLC lambda were manually diluted until the concentrations of the samples were at the low end of the initial measuring range. The results of the dilutions are compared to the upper limit of the calibration curve. When pre-reaction bit values (signals) for either assay are higher than the bit value of the upper end of the calibration curve, on either instrument, a new dilution and measurement are started automatically by the system.

Due to the built-in pre-reaction protocols on BN II and BN ProSpec, no hook effect was observed as false negatives when samples up to 27,100 mg/L for FLC kappa and up to 57,300 mq/L for FLC lambda were tested.

5.6.1.6 Specificity

The N Latex FLC kappa and lambda assays were evaluated for interference on BN Systems according to CLSI guideline EP7-A210. Following concentrations of listed endoqenous and exogenous substances were found to cause no interference up to the indicated concentrations:

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

Interferent	No Interference up to
Acetamidophenol	1 324 umol/L
Acetylsalicylic acid	3.62 umol/L
Amikacin	136.8 umol/L
Aminophylline Hydrate (Theophylline)	222 umol/L
Ascorbic acid	342 umol/L
Bilirubin conjugated	1 025 umol/L
Bilirubin unconjugated	618 umol/L
Caffeine	308 umol/L
Carbamazepine	127 umol/L
Chloramphenicol	155 umol/L
Chlordiazepoxide	33.3 umol/L
Chlorpromazine	6.3 µmol/L
Cimetidine	79.2 µmol/L
Creatinine	5 mg/dL
Dexamethasone	1.53 µmol/L
Dextran	60 g/L
Dextropropoxyphene	4.91 µmol/L
Diazepam	18 µmol/L
Digoxin	7.8 nmol/L
Erythromycin	81.6 µmol/L
Ethanol	100 mg/dL
Ethosuximide	1770 umol/L
Furosemide	181 µmol/L
Gentamicin	21 umol/L
Hemoglobin	10 g/L
Heparin Ammonium Salt	3 000 U/L
Heparin Lithium Salt	3 000 U/L
Heparin Sodium Salt	3 000 U/L
Ibuprofen	2 425 umol/L
Lidocaine	51.2 µmol/L
Lithium Chloride	3.2 mmol/L
Melphalan	4 000 ng/mL
Nicotine	6.2 µmol/L
Interferent	No Interference up to...
Penicillin	161 µmol/L
Pentobarbital	431 µmol/L
Phenytoin	198 µmol/L
Primidone	183 µmol/L
RF	2 000 IU/mL
Total Protein	143 g/L
Triglycerides	5 g/L
Urea	42.9 mmol/L
Uric acid	1.4 mmol/L
Valproic acid	3 467 µmol/L

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

5.7 Clinical Studies

5.7.1 Expected values / Reference interval

A reference interval study for N Latex FLC kappa and N Latex FLC assay was performed according to CLSI EP28-A3C, 'Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory'.

The reference intervals were determined from a US-population of 201 apparently healthy subjects. The reference intervals were calculated non-parametrically and represent the central 95 % range of the population.

The following reference intervals apply for serum and plasma samples from healthy adults:

2.5th-97.5th percentile
FLC kappa	8.24–28.9 mg/L
FLC lambda	9.10–32.6 mg/L

The calculation of the к/л ratios resulted in 0.88 (0.53 to 1.51) (median, 1.08 — 99.00 percentile).

The sponsor recommends that each laboratory should determine its own reference intervals since values may vary depending on the individual population studied.

{16}------------------------------------------------

5.7.2 Clinical Sensitivity and Specificity

A total of 342 samples were included in the clinical validation study for the N Latex FLC kappa and lambda assay. This validation set included 96 samples from Multiple Myeloma patients, 83 samples from AL Amyloidosis patients and 163 samples from non-myeloma patients with various clinical conditions: 24 polyclonal immunoglobulin stimulation: 16 Chronic Kidney Disease (CKD) and 123 other clinical conditions.

Clinical sensitivity and specificity summary of the N Latex FLC kappa and lambda ratio for Multiple Myeloma are shown in the table below:

		Clinical Diagnosis of Multiple Myeloma
		Positive	Negative	Total
N Latex FLC kappaand lambda ratio	Positive	92	5	97
	Negative	4	158	162
	Total	96	163	259

Clinical Sensitivity: 95.8 % (95 % Confidence Interval: 89.8 to 98.4 %) Clinical Specificity: 96.9 % (95 % Confidence Interval: 93.0 to 98.7 %)

Clinical sensitivity and specificity summary of the N Latex FLC kappa and lambda ratio for AL Amyloidosis are shown in the table below:

		Clinical Diagnosis of Amyloidosis
		Positive	Negative	Total
N Latex FLC kappaand lambda ratio	Positive	69	5	74
	Negative	14	158	172
	Total	83	163	246

Clinical Sensitivity: 83.1 % (95 % Confidence Interval: 73.7 to 89.7 %) Clinical Specificity: 96.9 % (95 % Confidence Interval: 93.0 to 98.7 %)

{17}------------------------------------------------

5.7.3 Method comparison with predicate device

152 serum samples from patients with monoclonal gammopathy were assayed by immunofixation (IFE), with N Latex FLC and in parallel with another commercially available particle-enhanced immunonephelometric method (comparison method).

	Comparison Method ⇒
N LatexFLC kappa ↓	<3.3 mg/L	3.3-19.4 mg/L	> 19.4 mg/L	total N
< 8.24 mg/L	6	11	0	17
8.24-28.9 mg/L	3	23	6	32
> 28.9 mg/L	0	1	102	103
total N	9	35	108	152

N Latex FLC kappa versus Comparison Method

overall agreement rate:

N Latex FLC lambda versus Comparison Method

ComparisonMethod ⇒N Latex FLClambda ↓	<5.7mg/L	5.7 - 26.3 mg/L	>26.3mg/L	total N
<9.10mg/L	16	10	0	26
9.10 - 32.6 mg/L	6	23	2	31
>32.6mg/L	0	10	85	95
total N	22	43	87	152

overall agreement rate:

5.8 Proposed Labeling

The labeling is adequate and satisfies requirements of 21 CFR Part 809.10.

5.9 Traceability

{18}------------------------------------------------

5.9.1 N FLC Standard SL

Calibration of the assay is traceable to an internal master calibrator; there is no international standard reference material.

5.9.2 N FLC Control SL1 and SL2

N FLC Controls are traceable to the master calibrator.

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).