N Latex FLC kappa, N Latex FLC lambda

K201496 · Siemens Healthcare Diagnostics Products GmbH · DFH · Oct 29, 2021 · Immunology

Device Facts

• Record ID: K201496
• Device Name: N Latex FLC kappa, N Latex FLC lambda
• Applicant: Siemens Healthcare Diagnostics Products GmbH
• Product Code: DFH · Immunology
• Decision Date: Oct 29, 2021
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5550
• Device Class: Class 2

Intended Use

N Latex FLC kappa and lambda are in-vitro diagnostic reagents for the quantitative determination of free light chains (FLC), type kappa or type lambda in human serum and EDTA plasma. N Latex FLC kappa and lambda assays are used: • as an aid in the diagnosis and monitoring of multiple myeloma (MM) on the BN Systems and Atellica® CH Analyzer. • as an aid in the diagnosis of immunoglobulin light-chain amyloidosis (AL) on the BN Systems and Atellica® CH Analyzer. • as an aid in the monitoring of immunoglobulin light-chain amyloidosis (AL) on the BN Systems. • as an aid in the evaluation of Monoclonal Gammopathy of Undetermined Significance (MGUS) on the BN Systems. Results of FLC measurements should always be interpreted in conjunction with other laboratory and clinical findings.

Device Story

In-vitro diagnostic reagents for quantitative determination of kappa and lambda free light chains (FLC) in human serum and EDTA plasma. Uses particle-enhanced nephelometric immunoassay; polystyrene particles coated with monoclonal mouse anti-human FLC antibodies agglutinate when mixed with patient samples. Scattered light intensity is proportional to FLC concentration; measured on BN Systems and Atellica CH Analyzer. Used by laboratory professionals to aid in diagnosis and monitoring of multiple myeloma, AL amyloidosis, and MGUS. Results interpreted by clinicians alongside other laboratory and clinical findings to assess disease status, treatment response, and progression.

Clinical Evidence

Multi-center study evaluated N Latex FLC assays for monitoring AL amyloidosis patients (n=241 for method comparison; n=222 for clinical status comparison). Compared N Latex FLC to Freelite predicate and clinical response criteria using three evaluation modes. Agreement rates between N Latex FLC and Freelite ranged from 45.5% to 90.0% depending on response criteria and evaluation mode. Concordance with clinical status for N Latex FLC (Evaluation Mode 2) showed 70.6% sensitivity and 96.1% specificity.

Technological Characteristics

Particle-enhanced nephelometric immunoassay. Reagents: polystyrene particles coated with monoclonal mouse anti-human FLC antibodies, supplementary reagents (mouse Ig, detergent), standard, and controls. Quantitative measurement via light scattering intensity. Instrument systems: BN II, BN ProSpec, Atellica CH Analyzer. No modification to kit components or principle of operation from previous clearances (K171742, K182098, K193047).

Indications for Use

Indicated for use as an aid in the diagnosis and monitoring of multiple myeloma (MM), diagnosis and monitoring of immunoglobulin light-chain amyloidosis (AL), and evaluation of Monoclonal Gammopathy of Undetermined Significance (MGUS) in human serum and EDTA plasma.

Regulatory Classification

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.

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)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health and Human Services logo on the left and the FDA logo on the right. The FDA logo features the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue text.

October 29, 2021

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

Re: K201496

Trade/Device Name: N Latex FLC kappa. N Latex FLC lambda 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 29, 2021 Received: August 2, 2021

## Dear Kerstin Koenigs:

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 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

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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/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); 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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

Ying (Katelin) Mao, Ph.D. Chief Division of Immunology and Hematology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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# Indications for Use

510(k) Number (if known) K201496

Device Name

N Latex FLC kappa and N Latex FLC lambda

#### Indications for Use (Describe)

N Latex FLC kappa and lambda are in-vitro diagnostic reagents for the quantitative determination of free light chains (FLC), type kappa or type lambda in human serum and EDTA plasma. N Latex FLC kappa and lambda assays are used:

• as an aid in the diagnosis and monitoring of multiple myeloma (MM) on the BN Systems and Atellica® CH Analyzer. • as an aid in the diagnosis of immunoglobulin light-chain amyloidosis (AL) on the BN Systems and Atellica® CH Analyzer.

· as an aid in the monitoring of immunoglobulin light-chain amyloidosis (AL) on the BN Systems.

· as an aid in the evaluation of Monoclonal Gammopathy of Undetermined Significance (MGUS) on the BN Systems.

Results of FLC measurements should always be interpreted in conjunction with other laboratory and clinical findings.

| Type of Use (Select one or both, as applicable) | |
|-------------------------------------------------|--|
|-------------------------------------------------|--|

|X Prescription Use (Part 21 CFR 801 Subpart D)

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

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Image /page/3/Picture/1 description: The image shows the logo for Siemens Healthineers. The word "SIEMENS" is written in teal, and the word "Healthineers" is written in orange below it. To the right of the word "Healthineers" is a graphic of orange dots.

# 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: K201496

### 1. Submitter

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

| Contact: | Kerstin Koenigs |
|----------------------|------------------------------------------|
| Email: | [e-mail] |
| Phone: | [phone] |
| Fax: | [phone] |
| Date of Preparation: | October 20, 2021 |

### 2. Device Information

| Trade Name: | N Latex FLC kappa<br>N Latex FLC lambda |
|-----------------------|---------------------------------------------------------------------------------------|
| Common or Usual Name: | Light Chain immunological test system |
| Classification Name: | Immunoglobulin (light chain specific) immunological<br>test system per 21CFR 866.5550 |
| Product Code: | DFH (kappa)<br>DEH (lambda) |
| Regulatory Class: | II |
| 510(k) Review Panel: | Clinical Immunology (82) |

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# 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

# 4. Device Description / Test Principle

N Latex FLC kappa and lambda are in-vitro diagnostic reagents for the quantitative determination of free light chains (FLC), type kappa or type lambda in human serum and EDTAplasma. N Latex FLC kappa and lambda assays are used as an aid in the diagnosis and monitoring of multiple myeloma (MM) and immunoglobulin light-chain amyloidosis (AL) and as an aid in the evaluation of Monoclonal Gammopathy of Undetermined Significance (MGUS). Monitoring of immunoglobulin light-chain amyloidosis (AL) and evaluation of MGUS are cleared for use only on the BN Systems.

The N Latex 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 free light chains. 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.

The devices in this submission have not materially changed since originally cleared under K171742. The purpose for this submission is to add monitoring of immunoglobulin light-chain amyloidosis (AL), on the BN Systems, to the intended use.

# 5. Intended Use / Indications for Use

### N Latex FLC kappa and N Latex FLC lambda assays

N Latex FLC kappa and lambda are in-vitro diagnostic reagents for the quantitative determination of free light chains (FLC), type kappa or type lambda in human serum and EDTA plasma.

N Latex FLC kappa and lambda assays are used:

- as an aid in the diagnosis and monitoring of multiple myeloma (MM) on the BN Systems ● and Atellica® CH Analyzer.
- as an aid in the diagnosis of immunoglobulin light-chain amyloidosis (AL) on the BN Systems and Atellica® CH Analyzer.
- . as an aid in the monitoring of immunoglobulin light-chain amyloidosis (AL) on the BN Systems.
- . as an aid in the evaluation of Monoclonal Gammopathy of Undetermined Significance (MGUS) on the BN Systems.

Results of FLC measurements should always be interpreted in conjunction with other laboratory and clinical findings.

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## Precaution

- . The performance of N Latex FLC kappa and lambda has not been thoroughly studied in IgM and Light Chain MGUS patients due to the low prevalence of these subtypes.
- . Patients with decreased renal function may have elevated FLC kappa and FLC lambda (Jacobs et al. N Latex FLC serum free light-chain assays in patients with renal impairment. Clin Chem Lab Med 2013, DOI 10.1515/cclm-2013-0864).
- . Sample populations excluded MGUS populations that were further diagnosed with a disease/disorder in subsequent testing with another medical device such as human immunodeficiency virus, hepatitis, and chronic lymphocytic leukemia. Thus, because the samples were enriched the specificity of the test may be inflated.

### 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).

### Special Conditions for Use:

For prescription use only.

Special instrument requirements:

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

## 6. Technical Characteristics

#### Similarities and Differences to the Predicate

A comparison of the similarities and differences between the proposed Siemens Healthcare Latex FLC kappa and lambda assays versus The Binding Site (TBS) Freelite Human Kappa Free Kit and Lambda Free Kit assays (predicate devices) is provided in the table below.

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| | Predicate Devices | Proposed Devices |
|-------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | The Binding Site | Siemens Healthcare<br>BN Systems |
| | Freelite® Human Kappa Free Kit<br>and Freelite® Human Lambda<br>Free Kit on the Siemens BN II | N Latex FLC kappa<br>N Latex FLC lambda |
| | K031016 | K171742, K182098, K193047 |
| Indications for<br>Use | Kappa: This kit is intended for the<br>quantitation of kappa free light chains<br>in serum and urine on the Siemens<br>BN II. Measurement of free light<br>chains aids in the diagnosis and<br>monitoring of multiple myeloma,<br>lymphocytic neoplasms,<br>Waldenstrom's macroglobulinemia,<br>AL amyloidosis, light chain deposition<br>disease and connective tissue<br>diseases such as systemic lupus<br>erythematosus in conjunction with<br>other laboratory and clinical findings.<br>Lambda: This kit is intended for the<br>quantitation of lambda free light<br>chains in serum and urine on the<br>Siemens BNTM II. Measurement of<br>free light chains aids in the diagnosis<br>and monitoring of multiple myeloma,<br>lymphocytic neoplasms,<br>Waldenstrom's macroglobulinemia,<br>AL amyloidosis, light chain deposition<br>disease and connective tissue<br>diseases such as systemic lupus<br>erythematosus in conjunction with<br>other laboratory and clinical findings. | N Latex FLC kappa and lambda are in-<br>vitro diagnostic reagents for the<br>quantitative determination of free light<br>chains (FLC), type kappa or type<br>lambda in human serum and EDTA<br>plasma.<br>N Latex FLC kappa and lambda assays<br>are used:<br>• as an aid in the diagnosis and<br>monitoring of multiple myeloma (MM)<br>on the BN Systems and Atellica® CH<br>Analyzer.<br>• as an aid in the diagnosis of<br>immunoglobulin light-chain amyloidosis<br>(AL) on the BN Systems and Atellica®<br>CH Analyzer.<br>• as an aid in the monitoring of<br>immunoglobulin light-chain amyloidosis<br>(AL) on the BN Systems.<br>• as an aid in the evaluation of<br>Monoclonal Gammopathy of<br>Undetermined Significance (MGUS) on<br>the BN Systems.<br>Results of FLC measurements should<br>always be interpreted in conjunction<br>with other laboratory and clinical<br>findings. |
| Sample Type | Human serum and urine | Human serum and EDTA plasma |
| Units | mg/L | Same |
| Technology | Nephelometry<br>Polystyrene particles coated with<br>polyclonal monospecific antibodies | Nephelometry<br>Polystyrene particles coated with<br>monoclonal antibodies |
| Traceability | Internal reference preparation | Internal Reference Plasma Pool |
| Calibrators | One level | Same |
| | Predicate Devices | Proposed Devices |
| | The Binding Site<br>Freelite® Human Kappa Free Kit<br>and Freelite® Human Lambda<br>Free Kit on the Siemens BN II | Siemens Healthcare<br>BN Systems<br>N Latex FLC kappa<br>N Latex FLC lambda |
| | K031016 | K171742, K182098, K193047 |
| Instrument<br>System | Siemens BN II System | Siemens BN II and BN ProSpec<br>Systems |
| Analytical<br>Measuring<br>Range<br>(Calibrator lot<br>dependent) | Kappa: 5.9 to 190 mg/L<br>Lambda: 5.0 to 160 mg/L | kappa: 3.4 to 110 mg/L<br>lambda: 1.9 to 60 mg/L |
| Reference<br>Interval | Kappa: 3.30 to 19.40 mg/L<br>Lambda: 5.71 to 26.30 mg/L<br>Ratio: 0.26 to 1.65 | kappa: 8.24 – 28.90 mg/L<br>lambda: 9.10 – 32.60 mg/L<br>Ratio: 0.53 to 1.51 |

## Comparison of Technological Characteristics

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510(k) Premarket Notification for Addition of immunoglobulin light-chain amyloidosis (AL) Monitoring Claim to N Latex FLC kappa and lambda

The differences between the predicate devices and proposed reagents 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.

# 7. Performance Data

Performance Data: Extended indication for monitoring of immunoqlobulin light-chain amyloidosis (AL).

See submissions K171742, K182098 and K193047 for previously documented analytical and clinical studies:

- Precision and Reproducibility ●
- Linearity / Assay Measuring Range ●
- Antigen Excess ●
- Stability ●
- Detection Capabilities ●
- . Analytical Specificity / Interferences
- Expected Values ●
- Clinical Specificity and Sensitivity ●
- Method Comparison to Predicate Devices ●

### 7.1 Performance data for monitoring of immunoglobulin light-chain amyloidosis (AL) patients

The Latex FLC assays were evaluated on BN II Systems in a multi-center study to evaluate performance in monitoring immunoglobulin light-chain amyloidosis (AL) patients. N Latex FLC kappa and lambda assays were compared to Freelite Kappa and Lambda assays (predicate

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devices) in a method comparison analysis. In addition, both methods were compared to the patient's "clinical response", taking into account all available clinical and laboratory information.

For each of the comparisons these three evaluation modes were applied to all immunoglobulin light-chain amyloidosis (AL) data sets, respectively:

- Evaluation Mode 11 ●
- Evaluation Mode 2² .
- . Evaluation Mode 33-4

Response evaluation was done by comparing changes between the initial sample draw and each consecutive blood draw independently, applying the rules as outlined in the table below.

| NCCN Response<br>Criteria | Serum M Protein<br>/ IFE | Evaluation Mode<br>1 | Evaluation Mode<br>2 | Evaluation Mode<br>3 |
|-----------------------------------------|------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| CR<br>(Complete Response) | Serum and Urine<br>IFE negative | FLC levels and<br>ratio normal | FLC levels and<br>ratio normal | FLC levels and<br>ratio normal |
| VGPR<br>(Very Good Partial<br>Response) | n/a | Reduction in the<br>dFLC to <40 mg/L | Reduction in the<br>dFLC to <40 mg/L | Reduction in the<br>dFLC to <40 mg/L |
| PR<br>(Partial Response) | n/a | dFLC ≥ 50 mg/L<br>A greater than<br>50% reduction in<br>the initial dFLC<br>value | dFLC ≥ 50 mg/L<br>A greater than<br>50% reduction in<br>the initial dFLC<br>value | dFLC ≥ 50 mg/L<br>1. A greater than<br>50% reduction<br>in the initial<br>dFLC value<br>2. For patients<br>with an initial<br>dFLC value of<br>less than 50<br>mg/L a low FLC<br>response is<br>indicated if<br>dFLC < 10<br>mg/L* |
| SD<br>(Stable Disease) | n/a | Less than a PR | Less than a PR | Less than a PR |
| PD<br>(Progressive<br>Disease) | From CR, any<br>detectable<br>monoclonal<br>protein<br>From PR, 50%<br>increase in serum<br>M protein to<br>> 0.5g/dL or 50% | From CR,<br>abnormal FLC<br>ratio (light chain<br>must at least<br>double)<br>From PR, Serum<br>FLC increase of<br>≥ 50% to iFLC | From CR,<br>abnormal FLC<br>ratio (light chain<br>must at least<br>double)<br>From PR, Serum<br>FLC increase of<br>≥ 50% to dFLC | From CR,<br>abnormal FLC<br>ratio (light chain<br>must at least<br>double)<br>From PR, Serum<br>FLC increase of<br>≥ 50% to dFLC |
| | increase in urine<br>M protein to > 200<br>mg/dL | > 100 mg/L | > 50 mg/L | > 20 and 20%<br>increase of<br>baseline |

## Therapy Response Criteria

* Note:-For dFLC <50 it is considered low-dFLC response and based on this citation? the sponsors combined partial response with low-dFLC response.

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The following studies were performed in support of an immunoglobulin light-chain amyloidosis (AL) monitoring claim:

#### Comparison to Predicate Devices

For comparison of the five response criteria (CR (Complete Response), VGPR (Very Good Partial Response), PR (Partial Response), SD (Stable Disease), PD (Progressive Disease)) three evaluation modes were applied to all immunoglobulin light-chain amyloidosis (AL) data sets. Siemens favors using evaluation mode 2.

Response categories were determined for free light chain testing by Siemens Healthcare N Latex FLC and TBS Freelite Kappa Free and Lambda Free. Response evaluation was done by comparing changes between the initial sample draw and each consecutive blood draw independently. The response levels obtained by the two different test systems were compared in 5x5 contingency tables and relative agreement calculated.

| Response<br>Criteria<br>based on<br>N Latex FLC<br>results | Response Criteria based on Freelite results | | | | | |
|------------------------------------------------------------|---------------------------------------------|------|---------------------|-------------------|------------------------|-------|
| | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease | Total |
| Complete<br>Response | 49 | 3 | 1 | 0 | 0 | 53 |
| VGPR | 19 | 48 | 9 | 2 | 0 | 78 |
| Partial<br>Response | 0 | 5 | 15 | 5 | 1 | 26 |
| Stable<br>Disease | 6 | 2 | 1 | 55 | 4 | 68 |
| Progressive<br>Disease | 0 | 0 | 1 | 4 | 11 | 16 |
| Total | 74 | 58 | 27 | 66 | 16 | 241 |

## Agreement of N Latex FLC versus Freelite (Evaluation Mode 1)

Agreement Rate and 95% Confidence Intervals of N Latex FLC versus Freelite (Evaluation Mode 1)

| Response<br>Criteria | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease |
|----------------------|----------------------|--------------|---------------------|-------------------|------------------------|
| Agreement<br>Rate | 66.2% | 82.8% | 55.6% | 83.3% | 68.8% |
| 95% CI<br>Bootstrap | 51.6 – 83.6% | 70.3 - 92.3% | 21.4 - 71.4% | 77.4 – 96.7% | 47.4 - 92.3% |

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## Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) and 95% Confidence Intervals of N Latex FLC versus Freelite (Evaluation Mode 1)

| | Result<br>(n / Total) | 95% CI Bootstrap |
|-----|-----------------------|------------------|
| PPA | 68.8% (11/16) | 47.4 – 92.3% |
| NPA | 97.8% (220/225) | 96.8 - 100.0% |

## Agreement of N Latex FLC versus Freelite (Evaluation Mode 2)

| Response<br>Criteria<br>based on<br>N Latex FLC<br>results | Response Criteria based on Freelite results | | | | | |
|------------------------------------------------------------|---------------------------------------------|------|---------------------|-------------------|------------------------|-------|
| | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease | Total |
| Complete<br>Response | 49 | 3 | 0 | 0 | 0 | 52 |
| VGPR | 17 | 45 | 14 | 3 | 0 | 79 |
| Partial<br>Response | 0 | 5 | 15 | 4 | 1 | 25 |
| Stable<br>Disease | 6 | 2 | 2 | 54 | 1 | 65 |
| Progressive<br>Disease | 0 | 0 | 2 | 3 | 15 | 20 |
| Total | 72 | 55 | 33 | 64 | 17 | 241 |

## Agreement Rate and 95% Confidence Intervals of N Latex FLC versus Freelite (Evaluation Mode 2)

| Response<br>Criteria | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease |
|----------------------|----------------------|--------------|---------------------|-------------------|------------------------|
| Agreement<br>Rate | 68.1% | 81.8% | 45.5% | 84.4% | 88.2% |
| 95% CI<br>Bootstrap | 53.6 – 86.3% | 67.4 – 91.1% | 15.0 – 60.0% | 76.5 – 95.4% | 70.0 – 100.0% |

## Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) and 95% Confidence Intervals of N Latex FLC versus Freelite (Evaluation Mode 2)

| | Result<br>(n / Total) | 95% CI Bootstrap |
|-----|-----------------------|------------------|
| PPA | 88.2% (15/17) | 70.0 – 100.0% |
| NPA | 97.8% (219/224) | 95.9 - 99.6% |

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| Response<br>Criteria | Response Criteria based on Freelite results | | | | | |
|------------------------------------|---------------------------------------------|------|---------------------|-------------------|------------------------|-------|
| based on N<br>Latex FLC<br>results | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease | Total |
| Complete<br>Response | 50 | 2 | 0 | 0 | 0 | 52 |
| VGPR | 19 | 48 | 7 | 2 | 1 | 77 |
| Partial<br>Response | 0 | 5 | 15 | 4 | 1 | 25 |
| Stable<br>Disease | 6 | 2 | 3 | 54 | 0 | 65 |
| Progressive<br>Disease | 0 | 0 | 1 | 3 | 18 | 22 |
| Total | 75 | 57 | 26 | 63 | 20 | 241 |

## Agreement of N Latex FLC versus Freelite (Evaluation Mode 3)

### Agreement Rate and 95% Confidence Intervals of N Latex FLC versus Freelite (Evaluation Mode 3)

| Response<br>Criteria | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease |
|----------------------|----------------------|--------------|---------------------|-------------------|------------------------|
| Agreement<br>Rate | 66.7% | 84.2 | 57.7 | 85.7% | 90.0% |
| 95% CI<br>Bootstrap | 52.0 – 83.9% | 71.2 – 93.3% | 23.1 – 76.0% | 77.0 – 96.1% | 77.8 – 100.0% |

### Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) and 95% Confidence Intervals of N Latex FLC versus Freelite (Evaluation Mode 3)

| | Result<br>(n / Total) | 95% CI Bootstrap |
|-----|-----------------------|------------------|
| PPA | 90.0% (18/20) | 77.8 – 100.0% |
| NPA | 98.2% (217/221) | 96.3 – 99.6% |

#### Comparison to Clinical Status

For comparison of the five response criteria (CR (Complete Response), VGPR (Very Good Partial Response), PR (Partial Response), SD (Stable Disease), PD (Progressive Disease)) three evaluation modes were applied to all immunoglobulin light-chain amyloidosis (AL) data sets. Siemens favors using evaluation mode 2.

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Response categories were determined for free light chain testing by Siemens Healthcare N Latex FLC and TBS Freelite Kappa Free and Lambda Free, respectively. Response evaluation was done by comparing changes between the initial sample draw and each consecutive blood draw independently.

The response levels obtained by the two different test systems were compared to the clinical response level provided by the physician taking into account all available clinical and laboratory information. Relative agreement between the clinical response (including further information in addition to serum testing) and the response level applying the clinical response criteria by either using N Latex FLC or the predicate device were calculated, also using 5x5 contingency tables, and compared.

Immunoglobulin light-chain amyloidosis (AL): Concordance Rate N Latex FLC Kappa and Lambda versus Clinical Status

| Response<br>Criteria | Response Criteria based on Clinical Status | | | | | |
|------------------------------------|--------------------------------------------|------|---------------------|-------------------|------------------------|-------|
| based on<br>N Latex FLC<br>results | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease | Total |
| Complete<br>Response | 19 | 8 | 10 | 6 | 1 | 44 |
| VGPR | 15 | 40 | 11 | 8 | 1 | 75 |
| Partial<br>Response | 0 | 4 | 15 | 6 | 1 | 26 |
| Stable<br>Disease | 0 | 4 | 17 | 35 | 5 | 61 |
| Progressive<br>Disease | 0 | 0 | 3 | 4 | 9 | 16 |
| Total | 34 | 56 | 56 | 59 | 17 | 222 |

## Concordance of N Latex FLC versus Clinical Status (Evaluation Mode 1)

### Concordance Rate and 95% Confidence Intervals of N Latex FLC versus Clinical Status (Evaluation Mode 1)

| Response<br>Criteria | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease |
|----------------------|----------------------|--------------|---------------------|-------------------|------------------------|
| Concordance<br>Rate | 55.9% | 71.4% | 26.8% | 59.3% | 52.9% |
| 95% CI<br>Bootstrap | 28.6 – 75.7% | 56.6 – 88.9% | 7.8 – 30.2% | 39.6 – 75.0% | 28.6 – 78.9% |

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## Sensitivity and Specificity and 95% Confidence Intervals of N Latex FLC versus Clinical Status (Evaluation Mode 1)

| | Result<br>(n / Total) | 95% CI Bootstrap |
|-------------|-----------------------|------------------|
| Sensitivity | 52.9% (9/17) | 28.6 – 78.9% |
| Specificity | 96.6% (198/205) | 95.2 — 99.5% |

## Concordance of N Latex FLC versus Clinical Status (Evaluation Mode 2)

| Response<br>Criteria<br>based on<br>N Latex FLC<br>results | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease | Total |
|------------------------------------------------------------|----------------------|------|---------------------|-------------------|------------------------|-------|
| Complete<br>Response | 18 | 8 | 10 | 6 | 1 | 43 |
| VGPR | 16 | 40 | 11 | 8 | 1 | 76 |
| Partial<br>Response | 0 | 4 | 15 | 6 | 0 | 25 |
| Stable<br>Disease | 0 | 4 | 17 | 34 | 3 | 58 |
| Progressive<br>Disease | 0 | 0 | 3 | 5 | 12 | 20 |
| Total | 34 | 56 | 56 | 59 | 17 | 222 |

## Concordance Rate and 95% Confidence Intervals of N Latex FLC versus Clinical Status (Evaluation Mode 2)

| Response<br>Criteria | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease |
|----------------------|----------------------|--------------|---------------------|-------------------|------------------------|
| Concordance<br>Rate | 52.9% | 71.4% | 26.8% | 57.6% | 70.6% |
| 95% CI<br>Bootstrap | 25.0 – 74.3% | 56.6 – 88.9% | 8.0 – 30.3% | 38.2 – 72.9% | 47.1 – 88.2% |

## Sensitivity and Specificity and 95% Confidence Intervals of N Latex FLC versus Clinical Status (Evaluation Mode 2)

| | Result<br>(n / Total) | 95% CI Bootstrap |
|-------------|-----------------------|------------------|
| Sensitivity | 70.6% (12/17) | 47.1 – 88.2% |
| Specificity | 96.1% (197/205) | 94.8 - 99.0% |

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| Response<br>Criteria | Response Criteria based on Clinical Status | | | | | |
|------------------------------------|--------------------------------------------|------|---------------------|-------------------|------------------------|-------|
| based on<br>N Latex FLC<br>results | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease | Total |
| Complete<br>Response | 18 | 8 | 10 | 6 | 1 | 43 |
| VGPR | 16 | 40 | 10 | 7 | 1 | 74 |
| Partial<br>Response | 0 | 4 | 15 | 6 | 0 | 25 |
| Stable<br>Disease | 0 | 4 | 18 | 34 | 2 | 58 |
| Progressive<br>Disease | 0 | 0 | 3 | 6 | 13 | 22 |
| Total | 34 | 56 | 56 | 59 | 17 | 222 |

## Concordance of N Latex FLC versus Clinical Status (Evaluation Mode 3)

## Concordance Rate and 95% Confidence Intervals of N Latex FLC versus Clinical Status (Evaluation Mode 3)

| Response<br>Criteria | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease |
|----------------------|----------------------|--------------|---------------------|-------------------|------------------------|
| Concordance<br>Rate | 52.9% | 71.4% | 26.8% | 57.6% | 76.5% |
| 95% CI<br>Bootstrap | 24.0 – 74.2% | 56.4 – 89.1% | 7.9 – 30.3% | 37.5 – 72.3% | 52.9 – 93.8% |

## Sensitivity and Specificity and 95% Confidence Intervals of N Latex FLC versus Clinical Status (Evaluation Mode 3)

| | Result<br>(n / Total) | 95% CI Bootstrap |
|-------------|-----------------------|------------------|
| Sensitivity | 76.5% (13/17) | 52.9 – 93.8% |
| Specificity | 95.6% (196/205) | 93.3 – 99.0% |

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# immunoglobulin light-chain amyloidosis (AL): Agreement Rate Freelite Kappa and Lambda versus Clinical Status

| Response<br>Criteria<br>based on<br>Freelite<br>results | | Response Criteria based on Clinical Status | | | | |
|---------------------------------------------------------|----------------------|--------------------------------------------|---------------------|-------------------|------------------------|-------|
| | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease | Total |
| Complete<br>Response | 21 | 14 | 12 | 13 | 1 | 61 |
| VGPR | 11 | 37 | 5 | 3 | 1 | 57 |
| Partial<br>Response | 2 | 2 | 15 | 7 | 0 | 26 |
| Stable<br>Disease | 0 | 3 | 23 | 32 | 4 | 62 |
| Progressive<br>Disease | 0 | 0 | 1 | 4 | 11 | 16 |
| Total | 34 | 56 | 56 | 59 | 17 | 222 |

# Concordance of Freelite versus Clinical Status (Evaluation Mode 1)

## Concordance Rate and 95% Confidence Intervals of Freelite versus Clinical Status (Evaluation Mode 1)

| Response<br>Criteria | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease |
|----------------------|----------------------|--------------|---------------------|-------------------|------------------------|
| Concordance<br>Rate | 61.8% | 66.1% | 26.8% | 54.2% | 64.7% |
| 95% CI<br>Bootstrap | 40.7 – 87.0% | 54.9 – 85.4% | 6.2 – 29.2% | 34.2 – 70.7% | 38.9 – 82.4% |

## Sensitivity and Specificity and 95% Confidence Intervals of Freelite versus Clinical Status (Evaluation Mode 1)

| | Result<br>(n / Total) | 95% CI Bootstrap |
|-------------|-----------------------|------------------|
| Sensitivity | 64.7% (11/17) | 38.9 - 82.4% |
| Specificity | 97.6% (200/205) | 94.8 - 99.5% |

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| Response<br>Criteria | Response Criteria based on Clinical Status | | | | | |
|---------------------------------|--------------------------------------------|------|---------------------|-------------------|------------------------|-------|
| based on<br>Freelite<br>results | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease | Total |
| Complete<br>Response | 20 | 14 | 12 | 13 | 1 | 60 |
| VGPR | 12 | 34 | 5 | 3 | 0 | 54 |
| Partial<br>Response | 2 | 5 | 16 | 7 | 2 | 32 |
| Stable<br>Disease | 0 | 3 | 21 | 31 | 4 | 59 |
| Progressive<br>Disease | 0 | 0 | 2 | 5 | 10 | 17 |
| Total | 34 | 56 | 56 | 59 | 17 | 222 |

## Concordance of Freelite versus Clinical Status (Evaluation Mode 2)

## Concordance Rate and 95% Confidence Intervals of Freelite versus Clinical Status (Evaluation Mode 2)

| Response<br>Criteria | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease |
|----------------------|----------------------|--------------|---------------------|-------------------|------------------------|
| Concordance<br>Rate | 58.8% | 60.7% | 28.6% | 52.5% | 58.8% |
| 95% CI<br>Bootstrap | 33.3 – 84.0% | 49.1 – 82.6% | 6.9 – 30.4% | 32.7 – 68.8% | 28.6 – 80.0% |

## Sensitivity and Specificity and 95% Confidence Intervals of Freelite versus Clinical Status (Evaluation Mode 2)

| | Result<br>(n / Total) | 95% CI Bootstrap |
|-------------|-----------------------|------------------|
| Sensitivity | 58.8% (10/17) | 28.6 – 80.0% |
| Specificity | 96.6% (198/205) | 94.2 - 99.5% |

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| Response<br>Criteria<br>based on<br>Freelite<br>results | Response Criteria based on Clinical Status | | | | | |
|---------------------------------------------------------|--------------------------------------------|------|---------------------|-------------------|------------------------|-------|
| | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease | Total |
| Complete<br>Response | 22 | 14 | 12 | 13 | 1 | 62 |
| VGPR | 11 | 36 | 5 | 3 | 1 | 56 |
| Partial<br>Response | 1 | 2 | 15 | 7 | 0 | 25 |
| Stable<br>Disease | 0 | 3 | 22 | 30 | 4 | 59 |
| Progressive<br>Disease | 0 | 1 | 2 | 6 | 11 | 20 |
| Total | 34 | 56 | 56 | 59 | 17 | 222 |

## Concordance of Freelite versus Clinical Status (Evaluation Mode 3)

## Concordance Rate and 95% Confidence Intervals of Freelite versus Clinical Status (Evaluation Mode 3)

| Response<br>Criteria | Complete<br>Response | VGPR | Partial<br>Response | Stable<br>Disease | Progressive<br>Disease |
|----------------------|----------------------|--------------|---------------------|-------------------|------------------------|
| Concordance<br>Rate | 64.7% | 64.3% | 26.8% | 50.8% | 64.7% |
| 95% CI<br>Bootstrap | 44.4 – 91.3% | 52.6 – 84.6% | 6.4 – 29.3% | 31.7 – 66.7% | 38.9 – 83.3% |

## Sensitivity and Specificity and 95% Confidence Intervals of Freelite versus Clinical Status (Evaluation Mode 3)

| | Result<br>(n / Total) | 95% CI Bootstrap |
|-------------|-----------------------|------------------|
| Sensitivity | 64.7% (11/17) | 38.9 – 83.3% |
| Specificity | 95.6% (196/205) | 93.4 - 99.0% |

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### Comparison to Predicate Devices using Follow-up Graphs

Patients were monitored and tested using N Latex FLC kappa and lambda and Freelite Kappa and Lambda after initiation of the first therapy phase and at intervals of ≥ one week with a total of at least three collections. Two graphs each were established for t…