The HYDRASHIFT 2/4 daratumumab test is intended for the qualitative detection of monoclonal proteins in human serum by immunofixation electrophoresis. The kits are to be used in conjunction with the HYDRAGEL IF kits and the semi-automated HYDRASYS 2 electrophoresis apparatus. The proteins, separated by electrophoresis on alkaline buffered agarose gels, are incubated with individual antisera that are specific against gamma (Ig G), alpha (Ig M) heavy chains, and kappa (free and bound) and lambda (free and bound) light chains, respectively. After removing the nonreacted proteins, the immunoprecipitates are stained with acid violet. The electrophoregrams are evaluated visually for the presence of specific reactions with the suspect monoclonal proteins. The HYDRASHIFT 2/4 daratumumab kits remove the daratumumab Ig G, Kappa interference and enable the visual evaluation of the presence of monoclonal proteins on the HYDRAGEL IF kits in patients who have received daratumumab therapy. For In Vitro Diagnostic Prescription Use Only.

The daratumumab Control is designed for quality control of the HYDRASHIFT daratumumab immunofixation procedure performed using the HYDRASYS 2 instrument. The daratumumab Control is designed for laboratory use. It should be used like a human serum. For In Vitro Diagnostic Prescription Use Only.

Device Description

HYDRASYS 2 is a semi-automated multi-parameter system for start-to finish agarose gel electrophoresis: application of samples, migration, drying, staining, destaining and final-stage drying.

Abnormal bands in serum protein electrophoregrams, primarily those in the beta globulin and gamma globulin zones, are always suspected to be monoclonal proteins (M-proteins, paraproteins, monoclonal immunoglobulins) and therefore, an indication of performing an Immunofixation technique to type and confirm the monoclonal gammopathies.

Daratumumab is a human therapeutic Ig G Kappa monoclonal antibody and as such, during the clinical monitoring of patients treated with daratumumab, this antibody simulates a band detected by serum protein electrophoresis and immunofixation in the gamma region. It can simulate an endogenous Ig G Kappa paraprotein.

Daratumumab CONTROL is a qualitative quality control for the assay.

The HYDRASHIFT daratumumab immunofixation procedure performed on HYDRAGEL IF 2/4 gel is based on the creation of a daratumumab / anti-daratumumab antibody complex and shifting it outside the gammaglobulins zone. With the HYDRASHIFT daratumumab procedure, the daratumumab / anti-daratumumab antibody complex is visualized in alpha-1 zone on Ig G and Kappa immunofixation tracks and then the interference is removed from the gamma zone.

AI/ML Overview

The HYDRASHIFT 2/4 daratumumab device is an immunofixation electrophoresis kit designed to remove daratumumab Ig G, Kappa interference, allowing for the visual evaluation of monoclonal proteins in human serum from patients treated with daratumumab. The study to prove the device meets acceptance criteria involved repeatability, reproducibility, and external comparative studies.

1. Table of Acceptance Criteria and Reported Device Performance

Acceptance Criteria	Reported Device Performance
Repeatability (Within-gel concordance)	100% concordant results for all 10 serum samples (1 normal, 9 with monoclonal components) run 4 times within the same gel.
Reproducibility (Between gels, lots, and instruments concordance)	100% concordant results for all 10 serum samples across 3 instruments, 3 lots, and spanning 3 working days.
External Comparative Study 1 Concordance (with/without HYDRASHIFT)	100% concordant results for 42 normal serum samples and 156 pathological serum samples when comparing results from HYDRAGEL 4 IF alone and HYDRASHIFT 2/4 daratumumab + HYDRAGEL 4 IF.
External Comparative Study 2 Concordance (with/without HYDRASHIFT)	100% concordant results for 38 normal serum samples and 134 pathological serum samples when comparing results from HYDRAGEL 4 IF alone and HYDRASHIFT 2/4 daratumumab + HYDRAGEL 4 IF.
Sensitivity (Detection limit of daratumumab/anti-daratumumab antibody complex)	0.3 q/L
Interference (with common interfering factors and drugs)	No interference detected with bilirubin (20 mg/dL), triglycerides (3.00 g/dL), hemoglobin (2 g/L), rheumatoid factor (2000 UI/mL), HAMA (Titer: 640), Pomalidomide (1 mg/L), Lenalidomide (4 mg/L), Dexamethasone (1 mg/L), Bortezomib (2 mg/L).

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

Repeatability Study: 10 different serum samples (1 normal, 9 with monoclonal components).
Reproducibility Study: 10 different serum samples (1 normal, 9 with monoclonal components).
External Comparative Study No. 1: 198 serum samples.
External Comparative Study No. 2: 172 serum samples (noted that the first 172 samples from study 1 were analyzed on both sites, implying potentially shared samples in part).

The data provenance is from "2 external studies performed in the USA." It is not explicitly stated whether the data was retrospective or prospective.

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

The document does not provide information on the number of experts used or their qualifications for establishing ground truth for the test set. The evaluation of electrophoregrams is noted as "evaluated visually," implying human interpretation, but specifics about the interpreters are absent.

4. Adjudication Method for the Test Set

The document does not explicitly describe an adjudication method (such as 2+1, 3+1) for the test set results. The concordance of results is reported without detailing how disagreements, if any, were resolved.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, and Effect Size

No MRMC comparative effectiveness study is reported in the provided text. The studies focus on the technical performance of the device (repeatability, reproducibility, and concordance with existing methods) rather than the improvement of human reader performance with or without AI assistance. The device is a "HYDRASHIFT" kit that removes interference for visual evaluation, not an AI-based interpretation system.

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

No standalone algorithm performance study was done. The device is a diagnostic kit that requires visual evaluation of electrophoregrams by a human. Its purpose is to prepare samples to remove interference for human readability, not to interpret the results automatically.

7. The Type of Ground Truth Used

The ground truth appears to be based on the established clinical characterization of the serum samples (e.g., "normal" or "pathological with monoclonal components"). For normal samples, it's the absence of monoclonal components. For pathological samples, it's the specific type of monoclonal component (e.g., Ig G, L + Ig A, K + Ig M, L). This would typically be confirmed by standard laboratory techniques and expert interpretation of immunofixation electrophoresis. The external comparative studies also use existing methods (HYDRAGEL 4 IF alone) as a comparison baseline.

8. The Sample Size for the Training Set

The document does not mention a "training set" as it is typically understood in the context of machine learning or AI. The studies performed are for device validation, focusing on performance characteristics such as repeatability, reproducibility, and comparison with predicate devices, rather than training a model.

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

Since there is no mention of a training set for an AI/algorithm, there is no information on how its ground truth would have been established. The ground truth for the validation studies (test sets) is based on the known characteristics of the serum samples and comparison with established methods.

Summary

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

January 11, 2018 Sebia, Inc Karen Anderson Director of Technical and Regulatory 1705 Corporate Drive, Suite 400 Norcross, Georgia 30093

Re: K172195

Trade/Device Name: HYDRASHIFT 2/4 daratumumab, daratumumab Control Regulation Number: 21 CFR 866.5510 Regulation Name: Immunoglobulins A, G, M, D, and E immunological test system Regulatory Class: Class II Product Codes: CFF. JJY Dated: December 07, 2017 Received: December 12, 2017

Dear Karen Anderson:

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 -S

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) K172195

Device Name HYDRASHIFT 2/4 daratumumab Daratbumumab Control

Indications for Use (Describe)

Assay:

Control:

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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510K SUMMARY (Summary of Safety and Effectiveness)

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

Submitter Name	Sebia, Inc.
Address	1705 Corporate Drive Suite 400Norcross, Georgia 30093, USA
Contact	Karen Anderson, Dir of Technical and RegulatoryPhone: 1-800-835-6497Fax: 770-446-8511Email: karen.anderson@sebia-usa.comYoussef Maakaroun, MD, Medical Affairs DirectorPhone 1-800-835-6497Fax 770-446-8511Email: youssef.maakaroun@sebia-usa.com
Date Prepared	January 4, 2018
Manufacturing	SebiaParc Technologique L\u00e9onard de VinciRue L\u00e9onard de Vinci,CP 8010 LISSES, 91008 EVRY CedexFRANCEPhone: (33) 1 69 89 80 80Fax: (33) 1 69 89 78 78
Product Name	HYDRASHIFT 2/4 daratumumabDaratumumab CONTROL
Common Name	Hydrashift daratumumab Serum Immunofixation
Product Regulation No.	21CFR sec. 866.5510
Product Codes	CFF
Device classification and PanelClassification	Class II , Immunology
Establishment Registration No.	8023024

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1. DEVICE DESCRIPTION

HYDRASYS 2 is a semi-automated multi-parameter system for start-to finish agarose gel electrophoresis: application of samples, migration, drying, staining, destaining and final-stage drying.

Daratumumab CONTROL is a qualitative quality control for the assay.

Reagents: REAGENTS AND MATERIALS SUPPLIED IN THE HYDRASHIFT 2/4 daratumumab KITS

ITEMS	PN 4639 (20 TESTS)	PN 4640 (40 TESTS)
Anti-daratumumab antiserum (ready to use)	1 vial, 0.4 mL	1 vial, 0.85 mL
Sample diluent (ready to use)	1 vial, 2,2 mL	1 vial, 2,2 mL
Green applicators (ready to use)	1 pack of 10 (15 teeth)	2 packs of 10 (15 teeth)

REAGENTS REQUIRED BUT NOT SUPPLIED

	SEBIA PRODUCT NUMBER
HYDRAGEL 2 or 4 IFAcid violet - Dynamic mask	4302, 4304 or 4381*
Antisera and Fixative for immunofixation IF -Dynamic maskor	4315
HYDRAGEL 2 or 4 IFAcid violet - Standard mask	4802, 4804 or 4881*
Antisera and Fixative for immunofixation IF -Standard maskand	4815
Daratumumab CONTROL	4765
DESTAINING SOLUTION	4540
HYDRASYS WASH SOLUTION	4541

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HYDRAGEL IF SAMPLE DILUENT	4588
FLUIDIL	4587
DTT DILUENT (IF / IT)	4589
BETA-MERCAPTOETHANOL (BME or2MERCAPTOETHANOL)	Not supplied by SEBIA

2. INDICATIONS FOR USE

The HYDRASHIFT 2/4 daratumumab test is intended for the qualitative detection of monoclonal proteins in human serum by immunofixation electrophoresis. The kits are to be used in conjunction with the HYDRAGEL IF kits and the semi-automated HYDRASYS 2 electrophoresis apparatus. The proteins, separated by electrophoresis on alkaline buffered agarose gels, are incubated with individual antisera that are specific against gamma (lg G), alpha (lg M) heavy chains, and kappa (free and bound) and lambda (free and bound) light chains, respectively. After removing the non-reacted proteins, the immunoprecipitates are stained with acid violet. The electrophoregrams are evaluated visually for the presence of specific reactions with the suspect monoclonal proteins. The HYDRASHIFT 2/4 daratumumab kits remove the daratumumab Iq G, Kappa interference and enable the visual evaluation of the presence of monoclonal proteins on the HYDRAGEL IF kits in patients who have received daratumumab therapy. For In Vitro Diagnostic Prescription Use Only.

The daratumumab Control is designed for the quality control of the HYDRASHIFT daratumumab immunofixation procedure performed using the HYDRASYS 2 instrument. The daratumumab Control is designed for laboratory use. It should be used like a human serum sample. For In Vitro Diagnostic Prescription Use Only.

3. TECHNOLOGICAL CHARACTERISTICS

4. SUBSTANTIAL EQUIVALENCE INFORMATION:

Predicate Device Name	Predicate Device510(k) number	Product Code	RegulationNo.
HYDRAGEL IF	K960669	CFF	866.5510

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Similarities between the candidate device (HYDRASHIFT 2/4 daratumumab) and the predicate device (HYDRAGEL IF) (Table A).

Similarities
Table A	HYDRASHIFT 2/4 daratumumabCandidate Device	HYDRAGEL IFPredicate Device ( K960669)
Intended use	The HYDRASHIFT 2/4 daratumumabtest is intended for the qualitativedetection of monoclonal proteins inhuman serum by immunofixationelectrophoresis. The kits are to be usedin conjunction with the HYDRAGEL IFkits and the semi-automatedHYDRASYS 2 electrophoresisapparatus. The proteins, separated byelectrophoresis on alkaline bufferedagarose gels, are incubated withindividual antisera that are specificagainst gamma (Ig G), alpha (Ig A) andmu (Ig M) heavy chains, and kappa (freeand bound) and lambda (free andbound) light chains, respectively. Afterremoving the non-reacted proteins, theimmunoprecipitates are stained withacid violet. The electrophoregrams areevaluated visually for the presence ofspecific reactions with the suspectmonoclonal proteins.TheHYDRASHIFT 2/4 daratumumab kitsremove the daratumumab Ig G, Kappainterference and enable the visualevaluation of the presence or absenceof monoclonal proteins on theHYDRAGEL IF kits in patients who havereceived daratumumab therapy. For InVitro Diagnostic Prescription Use Only.	The HYDRAGEL 1 IF, 2 IF, 4 IF and 9 IFkits are designed for detection ofmonoclonal proteins in human serumand urine by immunofixationelectrophoresis. The kits are used inconjunction with the semi-automatedHYDRASYS electrophoresis apparatus.The proteins, separated byelectrophoresis on alkaline bufferedagarose gels, are incubated withindividual antisera that are specificagainst gamma (Ig G), alpha (Ig A) andmu (Ig M) heavy chains, and kappa (freeand bound) and lambda (free and bound)light chains, respectively. After removingthe non-reacted proteins, theimmunoprecipitates are stained eitherwith acid violet or amidoblack. Theelectrophoregrams are evaluated visuallyfor the presence of specific reactionswith the suspect monoclonal proteins.
Assay Principle	Agarose Gel Electrophoresis	Agarose Gel Electrophoresis
Program	Same	Uses IF Program on Hydrasys 2
ReagentsGel kitAntisera kit	SameSame	HYDRAGEL IFAntisera and Fixative forimmunofixation IFNA
Differences
Table B	HYDRASHIFT 2/4 daratumumabCandidate Device	HYDRAGEL IFPredicate Device K960669)
Specimen Type	Human Serum	Human Serum, Human Urine
Reagents	Using anti-daratumumab antibody	No anti-daratumumab antibody
Daratumumab band	Removed from gamma zone into alpha zone	Remains in Gamma zone

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Table B. Differences between the predicate device (HYDRAGEL IF) and the candidate device (HYDRASHIFT 2/4 daratumumab) in (Table B).

5. Performance Data:

a) Repeatability

Ten (10) different serum samples, including 1 normal serum sample and 9 serum samples with monoclonal components, were run using the HYDRASHIFT 2/4 daratumumab procedure used in conjunction with each of the following kits: - HYDRAGEL 4 IF Acid violet Standard mask, -HYDRAGEL 4 IF Acid violet Dynamic mask.

Each sample was run 4 times within the same gel.

For each tested sample, all repeats gave 100 % of concordant results within gel.

Sample No.	Type	Within gel	Total analyses pergel
Sample No. 1	Normal	100% concordantresult	4
Sample No. 2	Ig G, L + Ig A, K + IgM, L	100% concordantresult	4
Sample No. 3	Ig G, K	100% concordantresult	4
Sample No. 4	Ig G, L	100% concordantresult	4
Sample No. 5	Ig A, K	100% concordantresult	4
Sample No. 6	Ig A, L	100% concordantresult	4
Sample No. 7	2 Ig M, K	100% concordantresult	4
Sample No. 8	Ig M, L	100% concordantresult	4
Sample No. 9	Kappa free	100% concordantresult	4
Sample No. 10	Lambda free	100% concordantresult	4

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b) Reproducibility between gels, between lots and between instruments

Ten (10) different serum samples, including 1 normal serum sample and 9 serum samples with monoclonal components, were run using the HYDRASHIFT 2/4 daratumumab procedure used in conjunction with each of the following kits : - HYDRAGEL 4 IF Acid violet Standard mask, - HYDRAGEL 4 IF Acid violet Dynamic mask.

This study was performed with 3 HYDRASYS 2 instruments and with 3 lots of HYDRASHIFT 2/4 daratumumab kit.

The normal sample was analyzed on 9 runs including one analysis per qel on 3 gels over 3 working days.

The samples with monoclonal components were analyzed on 9 runs with one analysis per gel, over 3 working days.

All samples gave 100 % of concordant results between gels on the 3 HYDRASYS 2 instruments and with 3 lots of HYDRASHIFT 2/4 daratumumab kit.

SampleNo.	Type	Instrument No. 1 / Lot No. 1			Instrument No. 2 / Lot No. 2			Instrument No. 3 / Lot No. 3			Total analysesper sample
		Day No. 1	Day No. 2	Day No. 3	Day No. 1	Day No. 2	Day No. 3	Day No. 1	Day No. 2	Day No. 3
1	Normal	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	27
2	Ig G, L +Ig A, K +Ig M, L	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	9
3	Ig G, K	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	9
4	Ig G, L	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	9
5	Ig A, K	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	9
6	Ig A, L	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	9
7	Ig M, K	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	9
8	Ig M, L	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	9
9	Kappafree	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	9
10	Lambdafree	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	100 %concordantresult	9

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c) External comparative studies

The results presented below have been obtained from 2 external studies performed in the USA. The external study No. 1 was performed on 198 samples and the external study No. 2 was performed on 172 samples. The first 172 samples have been analyzed on both sites.

External study No. 1

Comparative study No. 1 was performed on 198 serum samples analyzed with:

HYDRAGEL 4 IF Acid Violet Dynamic Mask kit,
HYDRASHIFT 2/4 daratumumab used in conjunction with HYDRAGEL 4 IF Acid Violet Dvnamic Mask kit.

With the HYDRAGEL 4 IF Acid Violet Dynamic Mask procedure, the daratumumab was visualized on G track and on Kappa track for 113 samples. For those samples, the HYDRASHIFT 2/4 daratumumab used in conjunction with HYDRAGEL 4 IF Acid Violet Dynamic Mask kit allows the shifting of the daratumumab.

For the other 85 samples, the characterization (normal or abnormal with monoclonal components) was the same between both procedures.

This study demonstrated 100 % concordant result:

For 42 normal serum samples: 100 % concordant result.
For the 156 pathological serum samples: 100 % concordant result.

External study No. 2

Comparative study No. 2 was performed on 172 serum samples analyzed with:

HYDRAGEL 4 IF Acid Violet Standard Mask kit, .
HYDRASHIFT 2/4 daratumumab used in conjunction with HYDRAGEL 4 IF Acid Violet - Standard Mask kit.

With the HYDRAGEL 4 IF Acid Violet Standard Mask procedure, the daratumumab was visualized on G track and on Kappa track for 98 samples, the HYDRASHIFT 2/4 daratumumab used in conjunction with HYDRAGEL 4 IF Acid Violet Standard Mask kit allows the shifting of the daratumumab.

For the other 74 samples, the characterization (normal or abnormal with monoclonal components) was the same between both procedures.

This study demonstrated 100 % concordant result:

For 38 normal serum samples: 100 % concordant result.
For the 134 pathological serum samples: 100 % concordant result.

d) Sensitivity

Six (6) serum samples, including 2 normal serum samples and 4 serum samples with different monoclonal components, added with daratumumab at different concentrations (final concentrations in sample between 0.1 and 3.0 g/L) were analyzed with the HYDRASHIFT 2/4 daratumumab procedure used in conjunction with each of the following kits: - HYDRAGEL 4 IF Acid Violet Standard Mask, - HYDRAGEL 4 IF Acid Violet Dynamic Mask.

The detection limit of daratumumab /anti-daratumumab /anti-daratumumab antibody complex visualized is 0.3 q/L.

e) Controls

It is recommended to run an assayed control serum (such as daratumumab CONTROL, SEBIA PN 4765) after each change of lot of a reagent.

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f) Interferences

The common interfering factors with the HYDRASHIFT 2/4 daratumumab procedure (bilirubin, triglycerides, hemoglobin and rheumatoid factor) were evaluated in studies based on the Clinical Laboratory Standards Institute (CLSI - USA) EP7-A2 guideline "Interference Testing in Clinical Chemistry; Approved Guideline – Second Edition". Additional inferference studies included: Human Anti-Mouse Antibody (HAMA, Pomalidomide, Lenalidomide, Dexamethasone, Bortezomib).

The results are summarized below

No interference with the HYDRASHIFT 2/4 daratumumab procedure was detected due to the serum sample's concentration of the following interfering factors tested at levels equal to the concentrations listed below :

Interfering factor	Concentration	Interfering factor/ Drug	Concentration
Bilirubin	20 mg/dL (342 μM)	Pomalidomide,	1 mg/L
Triglycerides	3,00 g/dL (34,5 mM)	Lenalidomide	4 mg/L
Hemoglobin	2 g/L (0,2 g/dL)	Dexamethasone	1 mg/L
Rheumatoid factor	2000 UI/mL	Bortezomib	2 mg/L
Human Anti-MouseAntibody (HAMA)	Titer : 640

q) Results

Interpretation, Interference and Limitations and Troubleshooting: See the instructions for use of the HYDRAGEL IF Standard mask or Dynamic mask kits.

A band on G and K immunofixation tracks outside the gammaqlobulins zone and in alpha-1 zone corresponds to the daratumumab / anti-daratumumab antibody complex. NOTE: Sometimes, the daratumumab complex is more faint in K track than in G track.

The HYDRASHIFT 2/4 daratumumab immunofixation removes the daratumumab lg G, Kappa interference and enables the visual evaluation of the presence or absence of monoclonal proteins on the HYDRAGEL IF kits, a negative test demonstrates the absence of monoclonal proteins in patients who have received daratumumab therapy.

The absence of any monoclonal protein on HYDRASHIFT 2/4 daratumumab immunofixation assay in either Alpha 2. Beta or Gamma zones is interpreted as a neqative assay for monoclonal protein.

The presence of a monoclonal protein ( or biclonal) on HYDRASHIFT 2/4 daratumumab immunofixation assay in either Alpha 2, Beta or Gamma zones is interpreted as a positive assay for monoclonal protein.

Some samples may give a faint band close to the application point due to a non-specific precipitation of proteins. This band may be visible on the G track and more particularly on the K track and cannot be confused with a monoclonal component

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6. IMWG Response Criteria

The current standard of practice for monitoring responses and relapses in multiple myeloma involve serum protein electrophoresis (SPEP) and Immunofixation (IF) (to determine complete response). International guidelines such as the National Comprehensive Cancer Network Clinical Practice Guidelines for Multiple Myeloma (NCCN) use reductions of monoclonal protein by SPEP and normalization of IFE to stratify response.

NCCN Guidelines Version 3.2017

*The Lancet Oncology 17 Kumar S, Paiva B, Anderson K, et al. International Myeloma Working Group consensus criteria for response and minimal response disease assessment in multiple myeloma, e328-46

*New criteria (2016) - International Myeloma Working Group
Stringent complete response	Complete response as defined below plus normal FLCratio** and absence of clonal cells in bone marrow biopsy byimmunohistochemistry (κ/λ ratio ≤4:1 or ≥1:2 for κ and λpatients, respectively, after counting ≥100 plasma cells)††
Complete response	Negative immunofixation on the serum and urine anddisappearance of any soft tissue plasmacytomas and <5%plasma cells in bone marrow aspirates
Very good partial response	Serum and urine M-protein detectable by immunofixation butnot on electrophoresis or ≥90% reduction in serum M-proteinplus urine M-protein level <100 mg per 24 h
Partial response	≥50% reduction of serum M-protein plus reduction in 24-hurinary M-protein by ≥90% or to <200 mg per 24 h;If the serum and urine M-protein are unmeasurable, a ≥50%decrease in the difference between involved and uninvolvedFLC levels is required in place of the M-protein criteria;If serum and urine M-protein are unmeasurable, andserumfree light assay is also unmeasurable, ≥50% reductionin plasma cells is required in place of M-protein, providedbaseline bone marrow plasma-cell percentage was≥30%. In addition to these criteria, if present at baseline, a≥50% reduction in the size (sum of the products of the maximalperpendicular diameters [SPD] of measured lesions)§§of soft tissue plasmacytomas is also required
Minimal response	≥25% but ≤49% reduction of serum M-protein and reduction in24-h urine M-protein by 50%-89%. In addition to theabove listed criteria, if present at baseline, a ≥50%reduction in SPD§§ of soft tissue plasmacytomas isalso required
Stable disease	Not recommended for use as an indicator of response; stabilityof disease is best described by providing thetime-to-progression estimates. Not meeting criteria forcomplete response, very good partial response, partialresponse, minimal response, or progressive disease
Progressive disease	Any one or more of the following criteria:Increase of 25% from lowest confirmed response value in oneor more of the following criteria:Serum M-protein (absolute increase must be ≥0.5 g/dL); SerumM-protein increase ≥1 g/dL, if the lowest M component was ≥5g/dL;Urine M-protein (absolute increase must be ≥200 mg/24 h);In patients without measurable serum and urine M-proteinlevels, the difference between involved and uninvolved FLClevels (absolute increase must be >10 mg/dL); In patientswithout measurable serum and urine M-protein levels andwithout measurable involved FLC levels, bone marrowplasma-cell percentage irrespective of baseline status(absolute increase must be ≥10%);Appearance of a new lesion(s), ≥50% increase from nadir inSPD§§ of >1 lesion, or ≥50% increase in the longest diameterof a previous lesion >1 cm in short axis;≥50% increase in circulating plasma cells (minimum of 200 cellsper µL) if this is the only measure of disease
Clinical relapse	Clinical relapse requires one or more of the following criteria:Direct indicators of increasing disease and/or end organdysfunction (calcium elevation, renal failure, anemia, lyticbone lesions [CRAB features]) related to the underlying clonalplasma-cell proliferative disorder. It is notused in calculation of time to progression or progression-freesurvival but is listed as something that can be reportedoptionally or for use in clinical practice;Development of new soft tissue plasmacytomas or bonelesions (osteoporotic fractures do not constitute progression);Definite increase in the size of existing plasmacytomas or bonelesions. A definite increase is defined as a 50%(and ≥1 cm) increase as measured serially by the SPD§§ of themeasurable lesion;Hypercalcemia (>11 mg/dL);Decrease in hemoglobin of ≥2 g/dL not related to therapy orother non-myeloma-related conditions;Rise in serum creatinine by 2 mg/dL or more from the start ofthe therapy and attributable to myeloma; Hyperviscosityrelated to serum paraprotein

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7. Conclusion:

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

Regulation Number and Section

§ 866.5510 Immunoglobulins A, G, M, D, and E immunological test system.

(a)
Identification. An immunoglobulins A, G, M, D, and E immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the immunoglobulins A, G, M, D, an E (serum antibodies) in serum. Measurement of these immunoglobulins aids in the diagnosis of abnormal protein metabolism and the body's lack of ability to resist infectious agents.(b)
Classification. Class II (performance standards).