EliA Rib-P is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Rib-P in human serum as an aid in the diagnosis of systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA Rib-P uses the EliA IgG method.

Device Description

EliA Rib-P is a semi-quantitative solid-phase fluoroenzymeimmunoassay, for the determination of autoantibodies against Rib-P. The EliA Rib-P test System is fully integrated and automated system which comprises of assay-specific reagents, EliA method-specific reagents, and general reagents.

Assav-Specific Reagents include:

EliA Rib-P Wells: coated with human recombinant ribosomal P-proteins P0, P1 . and P2 - 2 carriers (12 wells each), ready to use;
. EliA ANA 3 Positive Control 250 or 2500/5000: Human monoclonal antibodies in Tris buffer containing IgG antibodies to Ro52, Rib-P and RNA Pol III – 6 single use vials, 0.3 mL each, ready to use;
. EliA IgG/IgM/IgA Negative Control 250 or 2500/5000: Human blood preparation from healthy donors in PBS containing BSA, detergent and 0.095% sodium azide - 6 single-use vials, 0.3 mL each, ready to use;

EliA Method-Specific Reagents include:

EliA Sample Diluent: PBS containing BSA, detergent and 0.095% sodium azide . - 6 bottles, 48 mL each, ready to use; or 6 bottles, 400 mL each, ready to use;
I EliA IqG Conjuqate 50 or 200: ß-Galactosidase labeled anti-lgG (mouse monoclonal antibodies) in PBS containing BSA and 0.06% sodium azide - 6 wedge shaped bottles, 5 mL each, ready to use; or 6 wedge shaped bottles, 19 mL each, ready to use:
. EliA IgG Calibrator Strips: Human IqG (0, 4, 10, 20, 100, 600 uq/L) in PBS containing BSA, detergent and 0.095% sodium azide - 5 strips, 6 single-use vials per strip, 0.3 mL each, ready to use;
EliA IgG Curve Control Strips: Human IgG (20 ug/L) in PBS containing BSA, . detergent and 0.095% sodium azide - 5 strips, 6 single-use vials per strip, 0.3 mL each, ready to use:
EliA IgG Calibrator Well: coated with mouse monoclonal antibodies 4 carriers . (12 wells each), ready to use;

General Reagents include:

Development Solution: 0.01% 4-Methylumbelliferyl-β-D-galactoside, <0.0010% ■ preservative - 6 bottles (11 mL, 17 mL, or 112 mL each), sufficient for 6x >110, 6x >170, or 6x >1165 determinations;
I Stop Solution: 4% Sodium Carbonate - 6 bottles (65 mL, 119 mL, or 2800 mL each), sufficient for 6x >292, 6x >560, or 6x >13100 determinations;
트 Washing Solution Additive: detergent, preservative <0.13% - 6x 17.2 mL, 2x 86mL, or 4x 850 mL;
I Washing Solution Concentrate: phosphate buffer - 6x 80 mL, 2x 400 mL, or 1x 2800 mL.

AI/ML Overview

The provided document is a 510(k) Summary for the EliA Rib-P device, an in vitro semi-quantitative immunoassay for detecting IgG antibodies directed to Rib-P as an aid in diagnosing Systemic Lupus Erythematosus (SLE). This document does not describe a study involving an AI-enabled device or human-in-the-loop performance. Instead, it focuses on the analytical and clinical performance of a standalone laboratory diagnostic device and its substantial equivalence to a predicate device.

However, I can extract and structure the information related to acceptance criteria and the performance study of this specific diagnostic device in the format requested, even without AI-specific parameters.

Acceptance Criteria and Device Performance Study for EliA Rib-P

The EliA Rib-P device is an automated semi-quantitative solid phase fluoroenzymeimmunoassay for the measurement of IgG antibodies directed to Rib-P in human serum. This summary outlines the performance characteristics tested to demonstrate its substantial equivalence to a legally marketed predicate device, the Quanta Lite Ribosome P ELISA.

1. Table of Acceptance Criteria and the Reported Device Performance

Parameter	Acceptance Criteria (Implied by Study Design)	Reported Device Performance (EliA Rib-P)
Analytical Performance
Precision/Reproducibility	Inter-run, inter-instrument, and lot-to-lot variability to be within acceptable limits (typically low %CV). CLSI EP05-A3 guidelines followed.	Phadia 250: - Range of Total Imprecision %CV across 5 samples: 4.0% - 13.3% - Within-lab Imprecision %CV across 4 samples: 3.5% - 13.8% Phadia 2500/5000: - Range of Total Imprecision %CV across 5 samples: 5.7% - 18.4% (one sample had high CV, 18.4%)
Linearity/Reportable Range	Coefficient of determination (R²) close to 1.00, and slopes close to 1.00, across the measuring range. CLSI EP6-A guidelines followed.	Phadia 250: - R² values: 1.00 for all three dilution ranges. - Slopes: 0.95, 1.00, 1.00. Phadia 2500E: - R² values: 0.99, 1.00, 0.99 for the three dilution ranges. - Slopes: 1.03, 1.00, 1.02. "Linearity was shown for the entire measuring range."
Detection Limit	LoB, LoD, and LoQ to be established according to CLSI EP17-A2 guidelines (false positives and false negatives < 5%).	Harmonized across instruments: - LoB: 0.1 EliA U/mL - LoD: 0.5 EliA U/mL - LoQ: 1.9 EliA U/mL LoD determined with false positives (α) < 5% and false negatives (β) < 5%.
Analytical Specificity	No significant interference from common endogenous and exogenous substances at specified concentrations. CLSI EP07-ED3 and EP37-ED1 followed.	No interference observed from Bilirubin F (40 mg/dL), Bilirubin C (40 mg/dL), Haemoglobin (1000 mg/dL), Lipemic factor (2000 mg/dL), Rheumatoid factor (550 IU/mL), Ibuprofen (21.9 mg/dL), Losartan (1.14 mg/dL), Hydroxychloroquine (0.23 mg/dL), Azathioprine (0.26 mg/dL), Prednisone (0.01 mg/dL), Rituximab (109 mg/dL), Infliximab (26.4 mg/dL). Ratio of blank/spiked sample ranged from 0.90 – 1.10.
Assay Cut-Off	Clear distinction between negative, equivocal, and positive results based on a study of healthy donors and SLE patients.	Defined cut-offs: - Negative: < 7 EliA U/mL - Equivocal: 7-10 EliA U/mL - Positive: > 10 EliA U/mL
Comparison Studies
Method Comparison (vs. Predicate Device)	High Positive Percent Agreement (PPA), Negative Percent Agreement (NPA), and Total Agreement with the predicate device. CLSI EP09c-ED3 followed.	EliA Rib-P (equivocal considered negative): - PPA: 94.7% (95% CI: 82.3% - 99.4%) - NPA: 98.6% (95% CI: 96.4% - 99.6%) - Total Agreement: 98.1% (95% CI: 96.0% - 99.3%) EliA Rib-P (equivocal considered positive): - PPA: 100% (95% CI: 90.7% - 100%) - NPA: 88.1% (95% CI: 83.7% - 91.6%) - Total Agreement: 89.5% (95% CI: 85.6% - 92.6%)
Instrument Comparison	Strong correlation and agreement between different Phadia instrument series.	Regression analysis showed a slope of 0.94 (95% CI: 0.93 - 0.98) and an intercept of -0.76 (95% CI: -1.20 - -0.48) between Phadia 250 and Phadia 2500E.
Clinical Studies
Clinical Sensitivity and Specificity	Acceptable sensitivity for SLE and high specificity against other autoimmune and infectious diseases.	EliA Rib-P (equivocal considered positive): - Sensitivity (for SLE): 34.9% (95% CI: 27.2% - 43.3%) - Specificity (against disease controls): 99.3% (95% CI: 97.9% - 99.9%) EliA Rib-P (equivocal considered negative): - Sensitivity (for SLE): 28.1% (95% CI: 21.0% - 36.1%) - Specificity (against disease controls): 99.8% (95% CI: 98.7% - 100%)

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

Precision/Reproducibility:
- Phadia 250: 5 samples, tested in 252 replicates each across 3 lots and 3 instruments over 7 days.
- Within-Lab Imprecision: 4 samples, tested in 80 replicates each on 1 instrument over 20 days.
- Phadia 2500/5000 (E-module): 5 samples, tested in 84 replicates each on 3 instruments over 7 days.
- Provenance: Not explicitly stated, typically laboratory-prepared controls or banked human serum samples.
Linearity/Assay Reportable Range: 3 serum samples (diluted). Provenance not explicitly stated.
Detection Limit: 4 blank and 4 low-level samples (depleted IgG sera and prepared low-level samples) tested in 5-fold determination across 3 runs on Phadia 250 and Phadia 2500E.
Analytical Specificity (Interference): 3 serum samples (one negative, one equivocal, one high positive) spiked with interfering substances. Provenance not explicitly stated.
Assay Cut-Off: A cohort of 70 apparently healthy blood donors and 30 samples from SLE patients. Provenance not explicitly stated.
Method Comparison with Predicate Device: 323 patient samples. Provenance not explicitly stated, but implies diverse clinical samples covering the measuring range.
Instrument Comparison: 47 positive, 10 equivocal, and 28 negative samples. Provenance not explicitly stated.
Clinical Sensitivity and Specificity: 560 clinically defined serum samples from various diagnostic groups (146 SLE, 414 disease controls). Provenance not explicitly stated, but these are patient samples with confirmed diagnoses.
Expected Values/Reference Range: 638 apparently healthy subjects, equally distributed by age and gender from Caucasian, African American, Hispanic, and Asian populations obtained from a blood bank.

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

The document does not mention the use of experts to establish ground truth for the analytical performance characteristics. For clinical studies, the "ground truth" for patient samples was based on "clinically defined serum samples with a diagnosis" (e.g., systemic lupus erythematosus, Celiac disease, etc.). This typically implies diagnosis by medical professionals (e.g., rheumatologists, gastroenterologists, etc.) based on established diagnostic criteria, but the number and specific qualifications of these experts are not explicitly stated in this summary.

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

No explicit adjudication method is mentioned. The ground truth for clinical samples appears to be based on pre-existing clinical diagnoses.

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

This section is not applicable as the device is a standalone in vitro diagnostic immunoassay, not an AI-enabled device or an assist device for human readers. No MRMC study was conducted.

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

This device is a standalone algorithm/device in the sense that it performs automated semi-quantitative measurement of antibodies without real-time human interpretation impacting the measurement result. The assay directly measures the amount of antibody via fluorescence. The interpretation of results (negative, equivocal, positive) is based on predefined cut-offs.

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

Analytical ground truth: Based on reference materials, spiked samples, and dilution series with known concentrations or characteristics.
Clinical ground truth: "Clinically defined serum samples with a diagnosis" (e.g., SLE patients, various disease controls). This implies established clinical diagnoses, likely based on standard diagnostic criteria, which could involve expert clinical assessment, pathology, and other laboratory findings.

8. The Sample Size for the Training Set

This document does not specify a separate "training set" in the context of machine learning or AI. For a traditional immunoassay, method development and optimization would involve various experiments and sample sets, but these are not explicitly termed "training sets" here. The "Assay Cut-Off" study used 70 healthy blood donors and 30 SLE patients to define the cut-off, which could be considered a form of "training" for the interpretive criteria.

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

As noted above, a formal "training set" in the AI sense is not described. For the cut-off determination, the ground truth was established by using "apparently healthy blood donors" and "samples from SLE patients," indicating that these samples had known clinical statuses (healthy or diagnosed with SLE).

Summary

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September 13, 2021

Phadia AB % Sheryl Skinner Associate Director RA/QA Phadia US Inc. 4169 Commercial Avenue Portage, Michigan 49002

Re: K202540

Trade/Device Name: EliA Rib-P Regulation Number: 21 CFR 866.5100 Regulation Name: Antinuclear Antibody Immunological Test System Regulatory Class: Class II Product Code: MOA Dated: September 1, 2020 Received: September 2, 2020

Dear Sheryl Skinner:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

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

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal

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statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/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 (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Ying 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) K202540

Device Name EliA Rib-P

Indications for Use (Describe)

X Prescription Use (Part 21 CFR 801 Subpart D)

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

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510(k) Summary

This 510(k) Summary is prepared in accordance with the requirements of 21 CFR Part 807.92.

Premarket Notification 510(k) No: K202540

Date of Summary Preparation: September 3, 2021

Manufacturer:	Phadia ABRapsgatan 7PP.O. Box 6460751 37 Uppsala, Sweden
Distributor:	Phadia US Inc.4169 Commercial AvenuePortage, MI 49002
Company Contact Person:	Sheryl SkinnerAssociate Director, Regulatory and QualityPhadia US Inc.4169 Commercial Avenue(269) 568-3603sheryl.skinner@thermofisher.com

Proprietary and Established Device Name: EliA Rib-P

Regulatory Information:

Product Code:	MQA
Classification:	Class II
Regulation:	21 CFR 866.5100 – Antinuclear Antibody ImmunologicalTest System
Panel:	Immunology

Purpose of Submission:

New Device

Measurand:

IgG autoantibodies specific to Rib-P proteins

Type of Test:

Automated semi-quantitative solid phase fluoroenzymeimmunoassay

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Image /page/4/Picture/0 description: The image contains the logo for Thermo Fisher Scientific. The words "Thermo Fisher" are in red, with "SCIENTIFIC" in black below it. The logo is simple and modern.

Intended Use:

EliA Rib-P is intended for the in vitro semi-quantitative measurement of lgG antibodies directed to Rib-P in human serum as an aid in the diagnosis of systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA Rib-P uses the EliA IgG method.

Indication(s) for Use:

Same as intended use

Special Conditions for Use:

Rx - For Prescription Use Only

Special Instrument Requirements:

For use on the Phadia 250 instrument and the Phadia 2500 and Phadia 5000 instrument series (E-modules).

Device Description:

Assav-Specific Reagents include:

EliA Rib-P Wells: coated with human recombinant ribosomal P-proteins P0, P1 . and P2 - 2 carriers (12 wells each), ready to use;
. EliA ANA 3 Positive Control 250 or 2500/5000: Human monoclonal antibodies in Tris buffer containing IgG antibodies to Ro52, Rib-P and RNA Pol III – 6 single use vials, 0.3 mL each, ready to use;
. EliA IgG/IgM/IgA Negative Control 250 or 2500/5000: Human blood preparation from healthy donors in PBS containing BSA, detergent and 0.095% sodium azide - 6 single-use vials, 0.3 mL each, ready to use;

EliA Method-Specific Reagents include:

EliA Sample Diluent: PBS containing BSA, detergent and 0.095% sodium azide . - 6 bottles, 48 mL each, ready to use; or 6 bottles, 400 mL each, ready to use;
I EliA IqG Conjuqate 50 or 200: ß-Galactosidase labeled anti-lgG (mouse monoclonal antibodies) in PBS containing BSA and 0.06% sodium azide - 6 wedge shaped bottles, 5 mL each, ready to use; or 6 wedge shaped bottles, 19 mL each, ready to use:
. EliA IgG Calibrator Strips: Human IqG (0, 4, 10, 20, 100, 600 uq/L) in PBS containing BSA, detergent and 0.095% sodium azide - 5 strips, 6 single-use vials per strip, 0.3 mL each, ready to use;
EliA IgG Curve Control Strips: Human IgG (20 ug/L) in PBS containing BSA, . detergent and 0.095% sodium azide - 5 strips, 6 single-use vials per strip, 0.3 mL each, ready to use:

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EliA IgG Calibrator Well: coated with mouse monoclonal antibodies 4 carriers . (12 wells each), ready to use;
General Reagents include:
Development Solution: 0.01% 4-Methylumbelliferyl-β-D-galactoside, <0.0010% ■ preservative - 6 bottles (11 mL, 17 mL, or 112 mL each), sufficient for 6x >110, 6x >170, or 6x >1165 determinations;
I Stop Solution: 4% Sodium Carbonate - 6 bottles (65 mL, 119 mL, or 2800 mL each), sufficient for 6x >292, 6x >560, or 6x >13100 determinations;
트 Washing Solution Additive: detergent, preservative <0.13% - 6x 17.2 mL, 2x 86mL, or 4x 850 mL;
I Washing Solution Concentrate: phosphate buffer - 6x 80 mL, 2x 400 mL, or 1x 2800 mL.

Instrument System

EliA Rib-P is run on the Phadia 250 instrument and the Phadia 2500 and 5000 instrument series. The instruments are automated platforms for EliA test procedures from sample and reagent handling to the processing of results.

Substantial Equivalence

Quanta Lite Ribosome P ELISA, INOVA Diagnostics, Inc., (K981237)

	Similarities
Feature	Proposed DeviceEliA Rib-P	Predicate DeviceQuanta Lite Ribosome P ELISA
Intended Use	EliA Rib-P is intended for the invitro semi-quantitativemeasurement of IgG antibodiesdirected to Rib-P in humanserum as an aid in the diagnosisof systemic lupuserythematosus (SLE) inconjunction with other laboratoryand clinical findings. EliA Rib-Puses the EliA IgG method.	QUANTA Lite Ribosome P is anenzyme-linked immunosorbentassay (ELISA) for the semi-quantitative detection ofRibosome P antibodies inhuman serum. The presence ofRibosome P antibodies can beused in conjunction with clinicalfindings and other laboratorytests to aid in the diagnosis ofSystemic Lupus Erythematosus(SLE) and other relatedconnective tissue diseases.
Internal Controls	Positive and negative Controlprovided with EliA ANA 3Positive Control 250 and2500/5000 and EliA IgG/IgM/IgANegative Control 250 and2500/5000, resp.	Low Positive, High Positive andNegative Controlincluded in the kit

Comparison with Predicate Device:

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Thermoris SCIENTI

Traditional 510(k) for EliA Rib-P A.6 510(k) Summary_V6

Assay technique	ELISA	Same
Type of test	Semi-quantitative	Same
Sample Dilution(taking a 1%pipettingimprecision intoconsideration,this sampledilution isregarded as asimilarity)	1:100	1:101
Reported Unit	EliA U/mL (arbitrary)	Units (arbitrary)

	Differences
Feature	Proposed Device	Predicate Device
	EliA Rib-P	Quanta Lite Ribosome P ELISA
Antigen	Human recombinant P-proteinsP0, P1 and P2	Synthetic Ribosome P antigen
Instrumentation	EliA Rib-P uses the EliA IgGmethod on the instrumentsPhadia 250 and the E-modulesof the Phadia 2500 and Phadia5000 series.	ELISA-Reader needed
Reactiontemperature	37°C controlled	Room temperature, 20-26°C
Detectionantibody(conjugate)	IgG conjugate: anti-human IgGβ-Galactosidase (mousemonoclonal antibodies)	IgG conjugate: HRP IgGConjugate, (goat), anti-humanIgG
Signal	Fluorescence	Optical density
Calibration	6-point total IgG Calibration6 vials of human IgG atconcentrations of0 - 4 - 10 - 20 - 100 - 600µg/L	One-point calibration
Calibration curve	Option to store curve for up to28 days and run curve controlsin each assay for calibration	N/A
Interpretation ofresults	Negative < 7 EliA U/mLEquivocal 7-10 EliA U/mLPositive > 10 EliA U/mL	Negative < 20 UnitsWeak positive 20-39 UnitsModerate positive 40-80 UnitsStrong positive > 80 Units
Substrate	Development Solution 0.01 %4-Methylumbelliferyl-β-D-galactoside.	TMB Chromogen

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<0.0010% preservative*
*Preservative: mixture of 5-
chloro-2-methyl-2H-isothiazol-3-
one [EC no. 247-500-7] and 2-
methyl-2H-isothiazol-3-one [EC
no. 220-239-6] (3:1)

Standard/Guidance Document Referenced

CLSI EP25-A, Evaluation of Stability of In Vitro Diagnostic Reagents, ■ September 2009
I CLSI EP05-A3, Evaluation of Precision of Quantitative Measurement Procedures, September 2014
트 CLSI EP6-A, Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach. April 2003
. CLSI EP07-ED3, Interference Testing in Clinical Chemistry, September 2018
. CLSI EP37-ED1, Supplemental Tables for Interference Testing, September 2018
. CLSI EP09c-ED3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples, June 2018
. CLSI EP17-A2, Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures, June 2012
. CLSI EP28-A3c, Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory, October 2010

Test Principle

The EliA tests are fluorescence immunoassays for the detection and measurement of human antibodies based on EliA solid-phase components, which contain specific antigens for the antibodies to be measured.

The specific antigen for the antibodies to be detected is bound to the EliA solid phase component (EliA Well). The EliA wells are molded cups comparable to excised wells from a microtiter plate. They are made of polystyrene and are coated with the respective antigen. The wells are at the same time a holder of the coupled antigen for convenient automation and a reaction chamber with reaction/washing solution handling based on pipetting to add and aspiration to remove liquids. If present in the patient's specimen, antibodies to these proteins bind to their specific antigen. After washing away non-bound antibodies, enzyme-labeled antibodies against human IgG antibodies (EliA IgG Conjugate) are added to form an antibody-conjugate complex. After incubation, non-bound conjuqate is washed away, and the bound complex is incubated with a Development Solution. After stopping the reaction, the fluorescence in the reaction mixture is measured. The assay directly measures the amount of antibody of interest bound to the antigen coating the EliA well, therefore the higher the value of fluorescent signal detected by the instrument, the higher the amount of antibody bound and detected in the sample tested. To evaluate test results, the response for patient samples is compared directly to the response for calibrators.

Phadia AB, Rapsgatan 7P, P.O. Box 6460, 751 37 Uppsala, Sweden

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Performance Characteristics

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

To determine the precision of the assay on the Phadia 250 instrument and the Phadia 2500 and Phadia 5000 instrument series, the variability was assessed on 5 samples.

Three lots were used to determine the precision of the assay on Phadia 250 (totaling 252 replicate determinations per sample).

One lot was used to determine the precision of the assay on Phadia 2500E, which is a representative of the Phadia 2500 and Phadia 5000 instrument series.

EliA Rib-P on Phadia 250:

To determine the precision of the assay on the Phadia 250 instrument, the variability was assessed in a study with 21 runs by examining the samples in 252 replicates across 3 lots and 3 Phadia 250 instruments over 7 days. The data was calculated against the calibration curve from Day 1. The statistical evaluation was performed by Analysis of Variance. The results are given in the table below.

MeanEliAU/mL	Within-RunSD	Within-Run%CV	Between-RunSD	Between-Run%CV	Between-InstrumentSD	Between-Instrument%CV	Lot-to-LotSD	Lot-to-Lot%CV	TotalImprecisionSD	TotalImprecision%CV
2.8	0.2	5.9	0.2	6.6	0.2	8.5	0.1	5.0	0.4	13.3
7.8	0.3	3.2	0.3	4.1	0.3	4.2	0.0	0.0	0.5	6.7
9.5	0.4	3.8	0.3	2.9	0.5	5.3	0.4	3.9	0.8	8.1
56.8	1.8	3.2	2.9	5.1	1.0	1.8	0.6	1.0	3.6	6.4
316.4	9.3	2.9	7.5	2.4	2.6	0.8	3.0	1.0	12.5	4.0

Within-lab Imprecision

To determine the within-lab precision of the assay, the variability was assessed in a study with 40 runs by examining the samples in 80 replicates on 1 instrument over 20 davs. The data was calculated against the calibration curve from Day 1. The statistical evaluation was performed by Analysis of Variance. The results are given in the table below.

Mean(EliA U/mL)	Within-Run		Between-Run		Between-Day		Within-LabImprecision
	SD	%CV	SD	%CV	SD	%CV	SD	%CV
3.0	0.2	7.2	0.4	11.8	0.1	3.6	0.4	13.8
7.6	0.3	3.9	0.4	5.9	0.0	0.0	0.5	7.0
11.0	0.2	2.2	0.5	4.2	0.3	2.3	0.5	4.7
87.6	2.0	2.2	2.3	2.7	1.6	1.8	3.0	3.5

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EliA Rib-P on the Phadia 2500 and Phadia 5000 instrument series: To determine the precision of the assay on the of the Phadia 2500 and Phadia 5000 instrument series (E-module), the variability was assessed in a study with 21 runs by examining the samples in 84 replicates on 3 Phadia 2500E instruments over 7 days. The data was calculated against the calibration curve from Day 1. The statistical evaluation was performed by Analysis of Variance.

The results are given in the table below.

Mean		Within-Run		Between-Run		Between-Instrument		Total Imprecision
EliAU/mL	n	SD	%CV	SD	%CV	SD	%CV	SD	%CV
2.1	83*	0.2	8.1	0.4	16.5	0.0	1.0	0.4	18.4
7.1	84	0.4	5.4	0.5	6.6	0.1	1.9	0.6	8.7
8.8	84	0.5	5.5	0.8	8.7	0.4	4.3	1.0	11.2
48.7	84	1.5	3.1	1.7	3.4	1.7	3.4	2.8	5.7
362.1	84	31.6	8.7	20.9	5.8	0.0	0.0	37.9	10.5

One sample result is missing due to an instrument error.

b) Linearity/Assay Reportable Range:

3 serum samples were diluted in EliA Sample Diluent and tested on Phadia 250 and Phadia 2500E. The ratios of observed/expected values were calculated. The results are summarized below.

Phadia 250

Dilution Range(EliA U/mL)	Slope	Intercept	R²
45.1 - 452.6	0.95	9.01	1.00
2.7 - 122.9	1.00	1.28	1.00
0.5 - 32.2	1.00	0.24	1.00

Phadia 2500E

Dilution Range(EliA U/mL)	Slope	Intercept	R2
8.5 - 427.6	1.03	2.96	0.99
0.5 - 34.3	1.00	0.44	1.00
0.7 - 22.6	1.02	0.30	0.99

Linearity was shown for the entire measuring range.

Hook Effect/Over the Range Results:

Not applicable. Results above the upper limit of the measuring range are reported as ">403". No recommendations are made for dilution of samples outside measuring range in the Directions For Use.

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c) Traceability, Stability, Expected Values (Controls, Calibrators, or Methods): Traceability:

The IgG calibrators are traceable (via unbroken chain of calibrations) to the International Reference Preparation (IRP) 67/86 of Human Serum Immunoglobulins A, G and M from WHO. New batches of IgG calibrators are compared to a secondary standard (standardized with the IRP) or the IRP directly and adjusted accordingly to meet the correct concentration.

The instrument measures specific IgG concentrations in ug/L. By using a conversion factor given by the lot-specific code of the EliA test well, the results are automatically converted to EliA U/mL.

Stability:

Data for open and closed real-time stability and on-board stability of EliA IgG reagents and general EliA reagents on Phadia 250 as well as on the Emodule of the Phadia 2500 and Phadia 5000 series were already cleared with several other EliA tests, e.q. under K141375 (EliA M2 on Phadia 250). For the Phadia 2500 and Phadia 5000 instrument series, they were already cleared under K061165/A003 (EliA CCP).

Shelf-life:

The stability of EliA Rib-P Wells was evaluated with a real-time study. The results support stability of the test under the recommended storage of 2 – 8°C for up to 36 months.

On-board stability:

The on-board stability EliA Rib-P carriers (containing the antigen coated wells) was tested over 8 weeks using 3 positive and 2 negative samples only on the Phadia 250 instrument. As the storage conditions in the Emodule of the Phadia 2500 and Phadia 5000 series are similar to the Phadia 250, the results can also be used for stability claims for these instruments. The on-board stability for the Phadia 250 was determined to be 28 days at 2-8°C.

Open Stability:

Stability after first opening of the foil bag containing the EliA Rib-P wells was tested with a real-time study. According to this study, a shelf-life of 9 months at 2-8°C after first opening can be assigned to EliA Rib-P wells.

d) Detection Limit:

Four blank and four low level samples were measured with two different reagent sets (two lots of antigen wells). The four blank samples were created from depleted IqG sera, each diluted with EliA Sample Diluent. The blank samples and the low-level samples were assayed in three runs using two different sets of EliA Rib-P Well lots over three different days on a Phadia 250 and Phadia 2500E each in 5-fold determination.

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For each instrument type, the total number of combined observations for blank and low-level samples is 120 (60 per reagent set, 15 per sample and reagent set).

The results are summarized in the table below:

Instrument	LoB	LoD	LoQ
	EliA U/mL	EliA U/mL	EliA U/mL
Phadia 250	0.0	0.5	1.9
E-module of the Phadia 2500and Phadia 5000 series	0.1	0.4	1.1

A harmonized LoB of 0.1 EliA U/mL, LoD of 0.5 EliA U/mL, and LoQ of 1.9 EliA U/mL for the immunoassay was used.

The LoD for EliA Rib-P is 0.5 EliA U/mL, determined consistent with the quidelines in CLSI document EP17-A2 and with proportions of false positives (α) less than 5% and false negatives (β) less than 5%: based on 240 determinations with 120 blank and 120 low-level replicates per instrument type; and LoB of 0.1 EliA U/mL.

e) Analytical specificity:

Endogenous and Exogenous Interference:

A study was run to investigate whether high concentrations of potentially interfering substances in serum, like bilirubin, hemoglobin, lipemic factor, rheumatoid factor, Ibuprofen, Losartan, Hydroxychloroquine, Azathioprine, Prednisone, Rituximab and Infliximab adversely affect the results of the new device.

Three serum samples (one negative sample, one sample with a concentration within the equivocal range, and one high positive sample) were prediluted in EliA Sample Diluent and spiked with the different interfering substances or blank solution. The samples were tested in triplicates. A calibration curve was run in duplicate. The runs were repeated twice. One batch of EliA antigen wells and one batch of system reagents were used throughout the studies.

The ratio of blank/spiked sample ranged from 0.90 – 1.10 for EliA Rib-P. No interference was observed up to the concentrations listed in the table below:

Potential InterferingCompound	Concentration inundiluted sample
Bilirubin F	40 mg/dL
Bilirubin C	40 mg/dL
Hemoglobin	1000 mg/dL
Lipemic factor	2000 mg/dL
Rheumatoid factor	550 IU/mL

Phadia AB, Rapsgatan 7P, P.O. Box 6460, 751 37 Uppsala, Sweden

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Ibuprofen	21.9 mg/dL
Losartan	1.14 mg/dL
Hydroxychloroquine	0.23 mg/dL
Azathioprine	0.26 mg/dL
Prednisone	0.01 mg/dL
Rituximab	109 mg/dL
Infliximab	26.4 mg/dL

Reference Sera:

Externally defined sera should be measured according to their target values as mentioned by the institution CDC. Using EliA Rib-P, all 12 CDC samples were found according to their target.

f) Assay Cut-Off:

To define the cut-off, a study was performed using a cohort consisting of 70 apparently healthy blood donors and 30 samples from SLE patients. The samples were measured on a Phadia 250 instrument.

The cut-off was set as follows for EliA Rib-P:

<7 EliA U/mL	Negative
7-10 EliA U/mL	Equivocal
>10 EliA U/mL	Positive

In case of equivocal results, it is recommended to retest the patient after 8-12 weeks.

1. Comparison Studies:
- a) Method Comparison with Predicate Device:

A total of 323 patient samples with concentrations covering the measuring range were tested.

The samples were analyzed with the EliA Rib-P and Quanta Lite Ribosome P ELISA assay. The test was run in single determination and evaluated according to the Directions for Use. The results are summarized in the tables below:

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N = 323	Quanta LiteRibosome Ppositive:> 20 Units	Quanta LiteRibosome Pnegative:< 20 Units	Total
EliA Rib-P positive:> 10 EliA U/mL	36	4	40
EliA Rib-P negative:< 10 EliA U/mL	2	281	283
Total	38	285	323
	Calculation	Agreement (%)	95% CI
Positive PercentAgreement	$100% x 36 / 38$	94.7	82.3 - 99.4
Negative PercentAgreement	$100% x 281 / 285$	98.6	96.4 - 99.6
Total Agreement	$100% x (36 + 281) / 323$	98.1	96.0 - 99.3

EliA Rib-P: equivocal results considered negative

EliA Rib-P: equivocal results considered positive

N = 323	Quanta LiteRibosome Ppositive:> 20 Units	Quanta LiteRibosome Pnegative:< 20 Units	Total
EliA Rib-P positive:>7 EliA U/mL	38	34	72
EliA Rib-P negative:< 7 EliA U/mL	0	251	251
Total	38	285	323
	Calculation	Agreement (%)	95% CI
Positive PercentAgreement	$100% x 38 / 38$	100	90.7 - 100
Negative PercentAgreement	$100% x 251 / 285$	88.1	83.7 - 91.6
Total Agreement	$100% x (38 + 251) / 323$	89.5	85.6 - 92.6

b) Instrument Comparison:
Performance of EliA Rib-P was evaluated on the Phadia 250 and Phadia 2500E instrument using 47 positive, 10 equivocal and 28 negative samples. The samples were analyzed in single determination on one Phadia 250 and one Phadia 2500E instrument each. The regression analysis results are summarized as follows:

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	Intercept	Slope
Estimate	-0.76	0.94
95% Cl	-1.20 - -0.48	0.93 - 0.98

1. Clinical Studies:
- a) Clinical Sensitivity and Specificity:

560 clinically defined serum samples with a diagnosis from patients with systemic lupus erythematosus (n = 146), Celiac disease (n = 13), Crohn's disease (n = 12), CTD overlap Non-MCTD (n = 10), Dermatomyositis (n = 4), Polymyositis (n = 6), Graves' disease (n = 12), Primary antiphospholipid syndrome (n = 12), Primary biliary cirrhosis (n = 21), Sjögren's syndrome (n = 23), Type 1 Diabetes (n = 12), Ulcerative colitis (n = 11) cancer (n = 10), Mixed connective tissue disease (n = 10), Rheumatoid arthritis (n = 30), bacterial infections (n = 36), viral infections (n = 56), Hashimoto's disease (n = 10), Granulomatosis with Polyanqiitis (n = 4), Autoimmune Hepatitis (n = 16), Polymyalgia Rheumatica (n = 25), Systemic sclerosis, diffuse (n = 48) and Systemic sclerosis, limited (n = 33) were used to determine sensitivity and specificity of the assay.

The results are summarized in the tables below.

n=560	DiagnosticGroup	DiseaseControls	Total
Positive test≥ 7 EliA U/mL	51	3	54
Negative test< 7 EliA U/mL	95	411	506
Total	146	414	560

EliA Rib-P - equivocal results evaluated as positive:

Sensitivity (95% CI): 34.9% (27.2% - 43.3%) Specificity (95% CI): 99.3% (97.9% - 99.9%)

EliA Rib-P - equivocal results evaluated as neqative:

n=560	DiagnosticGroup	DiseaseControls	Total
Positive test> 10 EliA U/mL	41	1	42
Negative test≤ 10 EliA U/mL	105	413	518
Total	146	414	560

Sensitivity (95% CI): 28.1% (21.0 % - 36.1%)

Phadia AB, Rapsgatan 7P, P.O. Box 6460, 751 37 Uppsala, Sweden

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Specificity (95% CI): 99.8% (98.7% - 100%)

The table below shows the results for each clinical subgroup:

	total	positive	positive
Diagnostic Groups	n	n	%
Systemic lupus erythematosus	136	38	28%
Systemic lupus erythematosus withsecondary antiphospholipid syndrome	10	3	30%
Target disease (Total)	146	41
Celiac disease	13	0	0%
Crohn's disease	12	0	0%
CTD overlap Non-MCTD	10	0	0%
Dermatomyositis	4	0	0%
Polymyositis	6	0	0%
Graves' disease	12	0	0%
Primary antiphospholipid syndrome	12	0	0%
Primary Biliary Cholangitis	21	0	0%
Sjögren's syndrome	23	0	0%
Type 1 Diabetes	12	0	0%
Ulcerative colitis	11	0	0%
Varied Cancer	10	0	0%
Mixed connective tissue disease	10	0	0%
Rheumatoid arthritis	30	0	0%
Bacterial infections	36	0	0%
Viral infections	56	0	0%
Hashimoto's disease	10	0	0%
Granulomatosis with Polyangiitis	4	0	0%
Systemic sclerosis, diffuse	48	1	2%
Autoimmune Hepatitis	16	0	0%
Polymyalgia Rheumatica	25	0	0%
Systemic sclerosis, limited	33	0	0%
Disease Controls (Total)	414	1
Total	560

b) Other Clinical Supportive Data: Not applicable.

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1. Clinical Cut-Off: Same as assay cut-off.
1. Expected Values/Reference Range:

Antibody prevalence in autoimmune patients varies widely depending on disease area. The proportion of sera from a normal population found positive for the antinuclear antibodies covered by the EliA Rib-P test is below 1%. Expected values may vary depending on the population tested.

The frequency distribution for antinuclear antibodies was investigated in a group of apparently healthy subjects equally distributed by age and gender, using sera from Caucasian, African American, Hispanic and Asian population obtained from a blood bank.

The results are given in the table below:

Test	n	MedianEliA U/mL	95th percentileEliA U/mL	99th percentileEliA U/mL
EliA Rib-P	638	1.6	3.4	5.0

Proposed Labeling

The labeling is drafted in accordance with the requirements of 21 CFR Part 809.10.

Conclusion

All available data support that both immunoassays, the new device EliA Rib-P and its predicate device Quanta Lite Ribosome P ELISA perform substantially equivalent.

Regulation Number and Section

§ 866.5100 Antinuclear antibody immunological test system.

(a)
Identification. An antinuclear antibody immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoimmune antibodies in serum, other body fluids, and tissues that react with cellular nuclear constituents (molecules present in the nucleus of a cell, such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a liver disease), rheumatoid arthritis, Sjögren's syndrome (arthritis with inflammation of the eye, eyelid, and salivary glands), and systemic sclerosis (chronic hardening and shrinking of many body tissues).(b)
Classification. Class II (performance standards).