Access BNP on DxI 9000 Clinical Performance Evaluation: Specimen Testing and Clinical Concordance Study
Conditions: Heart Failure
Sponsor: Beckman Coulter, Inc.

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 1 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
DOCUMENT TITLE
ACCESS BNP ON DXI 9000 CLINICAL PERFORMANCE 
EVALUATION: SPECIMEN TESTING AND CLINICAL CONCORDANCE 
STUDY
PROJECT NAME
BNP FOR DXI 9000
PROJECT NUMBER
ID9016
PROTOCOL NUMBER
BNP-02-23
DHF DELIVERABLE NUMBER
2.7.2
REVISION
SPONSOR
BECKMAN COULTER, INC.
1000 LAKE HAZELTINE DRIVE
CHASKA, MN 55318
CONFIDENTIAL STATEMENT
This document contains information that is privileged or confidential and may not be disclosed 
unless such disclosure is required by Federal or State law or regulations. The information in this 
document and the results of the study may be disclosed only to those persons involved in the 
study who have a need to know, but all such persons must be instructed not to further 
disseminate this information to others.  These restrictions on disclosure will apply equally to all 
future information supplied to you and the future results of the study. 
2.7
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 2 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
STATEMENT OF COMPLIANCE
This study will be conducted in compliance with the protocol, the ethical principles that have their origin 
in the Declaration of Helsinki, the International Conference on Harmonization (ICH) consolidated 
Guideline E6 for Good Clinical Practice (GCP), and applicable regulatory requirement(s) listed below.
•
Code of Federal Regulations 21 CFR, Part 11
•
Code of Federal Regulations, Title 21, Part 50
•
Code of Federal Regulations 21 CFR, Part 54
•
Code of Federal Regulations, Title 21, Part 56
•
Code of Federal Regulations 21 CFR, Part 812.140, Subpart G
•
Code of Federal Regulations 45 CFR, Part 164
•
Code of Federal Regulations 45, CFR, Part 160
•
ETS No. 108: Convention for the Protection of Individuals with Regard to Automatic Processing 
of Personal Data
•
ETS No.164: Convention for the Protection of Human Rights and Dignity of the Human Being 
with Regard to the Application of Biology and Medicine: Convention on Human Rights and 
Biomedicine.
•
International Compilation of Human Research Protections
•
General Data Protection Regulation (GDPR) (EU) 2016/679
•
IVDR 2017/746
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 3 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
TABLE OF CONTENTS
TABLE OF CONTENTS ............................................................................................................................3
LIST OF ABBREVIATIONS.......................................................................................................................5
1.
KEY ROLES AND CONTACT INFORMATION.................................................................................6
1.1.
SPONSOR STUDY STAFF CONTACT INFORMATION.........................................................................................6
1.2.
SITE INVESTIGATOR CONTACT INFORMATION................................................................................................6
2.
STUDY SYNOPSIS...........................................................................................................................7
3.
INTRODUCTION...............................................................................................................................9
3.1.
BACKGROUND ............................................................................................................................................9
3.2.
STUDY PURPOSE........................................................................................................................................9
3.3.
SUBJECT RISK BENEFIT ANALYSIS.............................................................................................................10
3.3.1.
POTENTIAL RISKS.................................................................................................................................10
3.3.2.
POTENTIAL BENEFITS ...........................................................................................................................10
3.4.
INVESTIGATIONAL DEVICE DESCRIPTION ....................................................................................................10
3.5
COMPARATOR DEVICE DESCRIPTION
3.6.
INTENDED USE .........................................................................................................................................11
3.7.
TRAINING AND PROFICIENCY .....................................................................................................................12
3.8.
INVESTIGATIONAL SITES............................................................................................................................12
3.9.
STUDY DURATION.....................................................................................................................................12
4.
SOURCE DOCUMENTS AND ACCESS TO SOURCE DATA/DOCUMENTS ...............................12
5.
DATA HANDLING AND RECORD KEEPING.................................................................................13
5.1.
DATA MANAGEMENT RESPONSIBILITIES .....................................................................................................13
5.2.
DATA CAPTURE METHODS ........................................................................................................................13
5.3.
TYPES OF DATA........................................................................................................................................14
5.4.
STUDY RECORDS RETENTION....................................................................................................................14
6.
ETHICS/PROTECTION OF HUMAN SUBJECTS...........................................................................14
6.1.
ETHICAL STANDARD..................................................................................................................................14
6.2.
INSTITUTIONAL REVIEW BOARD/INDEPENDENT ETHICS COMMITTEE .............................................................14
6.3.
PARTICIPANT CONFIDENTIALITY.................................................................................................................14
7.
OBJECTIVES..................................................................................................................................15
7.1.
STUDY OBJECTIVES..................................................................................................................................15
7.2.
STUDY ENDPOINTS ...................................................................................................................................15
8.
STUDY DESIGN .............................................................................................................................15
9.
STUDY REQUIREMENTS ..............................................................................................................16
9.1.
FAMILIARIZATION TESTING.........................................................................................................................16
9.2.
INSTRUMENT MAINTENANCE AND QUALIFICATION........................................................................................17
9.3.
CALIBRATION............................................................................................................................................17
9.4
COMPARATOR METHOD………………………………………………………………………..…………………17
9.5.
QUALITY CONTROL ANALYSIS....................................................................................................................17
9.6.
QUALITY CONTROL ACCEPTANCE CRITERIA ...............................................................................................18
9.7.
REAGENT/PRODUCT HANDLING AND STORAGE...........................................................................................18
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 4 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
9.8.
SAMPLE HANDLING AND STORAGE.............................................................................................................19
9.9.
INVESTIGATIONAL PRODUCT ACCOUNTABILITY............................................................................................20
9.10.
PREMATURE STUDY TERMINATION OR SUSPENSION....................................................................................21
10.
STUDY FLOW CHART ...............................................................................................................21
11.
STUDY TESTING........................................................................................................................22
11.1.
SAMPLE/ISOLATE REQUIREMENTS .............................................................................................................22
11.2
COMPARATOR METHOD…………………………………………………………………………………………..21
11.3.
INVESTIGATIONAL METHOD........................................................................................................................22
12.
ASSESSMENT OF SAFETY AND PRODUCT QUALITY ...........................................................23
13.
SAFETY WARNINGS AND PRECAUTIONS ..............................................................................23
14.
CLINICAL SITE MONITORING...................................................................................................24
15.
REGULATORY AND ADMINISTRATIVE REQUIREMENTS......................................................24
15.1.
RESPONSIBILITY OF INVESTIGATOR............................................................................................................24
15.2.
RESPONSIBILITY OF SPONSOR...................................................................................................................25
15.3.
PROTOCOL DEVIATIONS ............................................................................................................................26
15.4.
STUDY SPECIFIC OR UNIQUE NONCOMPLIANCE CONCERNS...........................................................................26
15.5.
CHANGES TO PROTOCOL ..........................................................................................................................26
15.6.
RECONCILIATION OF STUDY MATERIAL, SUPPLIES AND SAMPLES.................................................................27
16.
STATISTICAL CONSIDERATIONS ............................................................................................27
17.
STUDY MATERIALS...................................................................................................................28
18.
SUPPLEMENTAL MATERIALS PROVIDED ..............................................................................29
19.
LITERATURE REFERENCES ....................................................................................................30
APPENDIX A: INVESTIGATOR PROTOCOL SIGNATURE PAGE........................................................31
APPROVALS ..........................................................................................................................................38
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 5 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
List of Abbreviations
TERM
DEFINITION
A2
Access 2 Immunoassay System
AMR
Analytical Measuring Range
BNP
Brain natriuretic peptide
C
Celsius
CKD
Chronic Kidney Disease
CRF
Case Report Form
CLSI
Clinical and Laboratory Standards Institute
DxI 9000
DxI 9000 Access Immunoassay Analyzer
eCRF
Electronic Case Report Form
eTMF
Electronic Trial Master File
ED
Emergency Department
FDA
U.S. Food and Drug Administration
GCP
Good Clinical Practice
HIPAA
Health Insurance Portability and Accountability Act
HF
Heart Failure
ICH
International Conference on Harmonization
IFU
Instructions for Use
IRB
Institutional Review Board
IUO
Investigational Use Only
IVD
In Vitro Diagnostic
NP
Natriuretic peptide
NPA
Negative Percent Agreement
NT-proBNP
N-terminal-pro hormone BNP 
OUS
Outside of United States
PPA
Positive Percent Agreement
QC
Quality Control
RLU
Relative light units
SOP
Standard Operating Procedure
US
United States
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 6 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
1. Key Roles and Contact Information
1.1. Sponsor Study Staff Contact Information
A list of the names and titles of Sponsor representatives is to be provided to the site as a separate 
document. Sponsor is Beckman Coulter, Inc at 1000 Lake Hazeltine Drive, Chaska, MN 55318. 
Main contact: (952) 448-4848. 
1.2.  Site Investigator Contact Information
A list of the names and titles of clinical investigators and the address and telephone number of 
study sites will be maintained in the sponsor master files and provided to investigators in the 
Investigator Site File. Beckman Coulter, Inc. will maintain an updated list, providing updates as 
required to the Investigator Site Files.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 7 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
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2.  Study Synopsis
Full Title
Access BNP on DxI 9000 Clinical Performance Evaluation: Specimen Testing 
and Clinical Concordance Study
Short Title
ACCESS BNP CLINICAL PERFORMANCE EVALUATION: TESTING AND CLINICAL 
CONCORDANCE STUDY
Protocol Number
BNP-02-23
Protocol Version
Study Sponsor
Beckman Coulter, Inc. 1000 Lake Hazeltine Dr. Chaska, MN 55318
Study Purpose
The overall aim of the research is to establish clinical performance of the 
Access BNP Assay (item number D06227) on the DxI 9000 Access 
Immunoassay Analyzer (P/N C11137) on the intended use population and 
establish clinical concordance to Access BNP Assay (item 98200, original 
formulation) on the Access 2 Immunoassay system (P/N 81600N).
The purpose of the protocol is to perform specimen testing (samples collected 
under the Access BNP Clinical Enrollment Protocol) to support clinical 
validation and the IUO Access BNP Assay’s regulatory approval on the DxI 
9000 Access Immunoassay Analyzer (P/N C11137). In addition, this protocol 
tests the samples using Access BNP assay (item number 98200, original 
formulation) on the Access 2 Immunoassay System (P/N 81600N) to perform 
a clinical concordance analysis to support global submissions, product launch 
and customer support materials. 
Investigational 
Material/Product
IUO Access BNP for DxI 9000 Access Immunoassay Analyzer
Reagent Pack (item number D06227)
Current State of the 
Art
Clinical practice guidelines3,4,6 demonstrate the generally accepted state of the 
art as an aid in the diagnosis of congestive heart failure (also referred to as 
heart failure), assessment of severity of heart failure, and risk stratification of 
patients with heart failure, and in risk stratification of patients with acute 
coronary syndromes with other recommended tests.3,4,6 The assessment of 
literature demonstrates that the currently marketed Access BNP assay is a 
useful tool to accurately measure BNP levels.
Study Objectives
The objective is to validate established diagnostic cutoff (rule-out) and assess 
HF severity on the IUO Access BNP Assay using the DxI 9000 Access 
Immunoassay Analyzer (P/N C11137). 
A method concordance analysis will be conducted between the IUO Access 
BNP (item number D06227) for DxI 9000 Access Immunoassay Analyzer (P/N 
C11137) and the Access BNP assay (item number 98200; original 
formulation) for Access 2 Immunoassay System (P/N 81600N).
2.7
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 8 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Study Design
•
This is a typical diagnostic device evaluation study with a single arm and 
no active intervention. 
•
A method concordance analysis will be conducted between the IUO 
Access BNP assay (item number D06227) for DxI 9000 Access 
Immunoassay Analyzer (P/N C11137) and the Access BNP assay (Item 
number 98200, original formulation) for Access 2 Immunoassay System 
(P/N 81600N).
•
A minimum of three (3) calibrator lots and a minimum of three (3) IUO 
reagent pack lots (Item number D06227) for DxI 9000 and three (3) IVD 
reagent pack lots (item 98200, original formulation) for Access 2 will 
be used in this study.
•
Each site will be provided a minimum of one (1) unique calibrator lot and a 
minimum of three (3) individual IUO Access BNP assay reagent pack lots 
(Item number D06227) for DxI 9000 from Sponsor. The IUO reagent lots 
will be delivered at designated time points determined by Sponsor.   
•
Each site will be provided a minimum of one (1) unique calibrator lot and a 
minimum of three (3) individual IVD Access BNP assay reagent pack lots 
(item 98200, original formulation) for Access 2 from Sponsor. The 
reagent lots will be delivered at designated time points determined by 
Sponsor.   
Inclusion Criteria
•
There are no inclusion criteria. Samples tested in this protocol are collected 
under BNP-05-24.
Exclusion Criteria
•
Hemolysis observed 4+ per CLSIC56A grading scale.
•
Icterus (bilirubin) observed 4+ per CLSIC56A grading scale. 
Target Population/ 
Sample Size
Specimens collected under BNP-05-24 protocol (titled: Access BNP Clinical 
Enrollment Study) are used. 
Approximately 1100 K2 EDTA samples with concentrations that span the 
analytical measuring range (AMR) are used in this testing study.
Site Requirements
A minimum of one (1) and approximately three (3) external testing sites that 
have a DxI 9000 Access Immunoassay Analyzer (P/N C11137) and Access 2 
Immunoassay System (P/N 81600N).  
Study Duration
The study duration for the clinical performance evaluation testing study is 
estimated to last approximately 12 months for sample testing and an estimated 
3 additional months for data analysis. 
Statistical Methods
All Statistical analysis will be performed by the Sponsor in accordance with 
CLSI EP12 guideline. A separate statistical analysis plan will detail the analysis 
methods used.
Primary endpoints of the study will be diagnostic accuracy between final 
clinical site diagnosis (HF and Non-HF) and IUO BNP assay testing results and 
severity assessment utilizing NYHA classification. 
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 9 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Method concordance will be established using the valid testing data from 
the investigational device Access BNP Assay for DxI 9000 Access 
Immunoassay Analyzer (P/N C11137) against the currently marketed 
Access BNP Assay for Access 2 Immunoassay System (P/N 81600N). 
Positive percent agreement (PPA) and Negative percent agreement (NPA) 
will be calculated using the data. 
3. Introduction
3.1.
Background
Heart Failure (HF) is a complex medical condition that often exists with other comorbidities.  
Further, symptoms of HF are nonspecific making HF diagnosis difficult. B-type natriuretic peptide 
biomarkers (BNP and NT-proBNP) are standard-of-care biomarkers measured at patient 
presentation in the emergency department (ED) for patients presenting with signs and symptoms 
suggestive of heart failure. The release of natriuretic peptides (NP) appears to be in direct 
proportion to ventricular volume expansion and pressure overload. The plasma concentrations of 
both hormones are increased in patients with asymptomatic and symptomatic left ventricular 
dysfunction, permitting their use in supporting a clinical diagnosis of HF (but not used alone for 
this purpose). NPs are well established for use in diagnosis of patients with HF and are included 
in standard of care guidelines for both the American College of Cardiology Foundation/American 
Heart Association (ACCF/AHA) and European Society of Cardiology (ESC).1,2,3,4 
In the Breathing Not Properly Study, B-type natriuretic peptide test accurately diagnosed HF in 
patients presenting to the ED with dyspnea, with a sensitivity of 90% and specificity of 76% at a 
cutoff of 100 pg/mL.5 The current commercialized Access BNP assay (primary predicate) for the 
Beckman Coulter Access Family of Immunoassay Systems has similarly established that BNP 
results less than or equal to 100 pg/mL are representative of normal values in patients without heart 
failure.
The Access BNP assay is being redesigned for transference to the DxI 9000 Access Immunoassay 
Analyzer (P/N C11137).
Specimen collected under BNP-05-24 “Access BNP Clinical Subject Sample Collection Enrollment 
Study Protocol” are to be tested on the investigational-use-only (IUO) Access BNP on Beckman 
Coulter DxI 9000 Access Immunoassay Analyzer (P/N C11137). 
3.2.  Study Purpose
The overall aim of the research is to establish clinical performance of the Access BNP Assay (item 
number D06227) on the DxI 9000 Access Immunoassay Analyzer (P/N C11137) on the intended 
use population and establish clinical concordance to Access BNP Assay (item 98200, original 
formulation) on the Access 2 Immunoassay system (P/N 81600N).
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 10 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
The purpose of the protocol is to perform specimen testing (samples collected under the Access 
BNP Clinical Enrollment Protocol) to support clinical validation and the IUO Access BNP Assay’s 
regulatory approval on the DxI 9000 Access Immunoassay Analyzer (P/N C11137). In addition, this 
protocol tests the samples using Access BNP assay (item number 98200, original formulation) 
on the 
Access 2 Immunoassay System to perform a clinical concordance analysis to support global 
submissions, product launch and customer support materials.
3.3. Subject Risk Benefit Analysis
3.3.1.
Potential Risks 
There are no potential known risks to the subjects during the testing phase of the clinical study.
The testing will be performed using de-identified clinical samples that will be supplied to the 
testing sites by the Sponsor. The testing is performed on samples containing only a unique 
sample identification number. Subject information such as names or addresses are not 
included. Protected health information (PHI) will not be revealed to the sponsor and will not be a 
part of the barcode label.
3.3.2.     Potential Benefits
There is no direct benefit to the subject from participating in the study. 
Potential benefits to the subject may include knowing that their contribution could benefit 
medical science in the future and could facilitate the development of accurate blood tests for 
detecting and monitoring disease. No results obtained from any analyses will be used to 
determine or alter the patient’s treatment or care.
3.4.
 Investigational Device Description
IUO Access BNP for DxI 9000 Reagent Pack Description (item number D06227):
The Access BNP test is a two-site immunoenzymatic (“sandwich”) assay. A sample is added to a 
reaction vessel with mouse monoclonal anti-human BNP antibody-alkaline phosphatase 
conjugate and paramagnetic particles coated with mouse Omniclonal anti-human BNP antibody. 
BNP in human plasma binds to the immobilized anti-BNP on the solid phase, while the mouse 
anti-BNP conjugate reacts specifically with bound BNP.
After incubation, materials bound to the solid phase are held in a magnetic field while unbound 
materials are washed away. Then, the chemiluminescent substrate is added to the vessel and 
light generated by the reaction is measured with a luminometer. The light production is directly 
proportional to the concentration of analyte in the sample. Analyte concentration is automatically 
determined from a stored calibration. 
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 11 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
3.5.  Comparator Device Description
The comparator device(s) are:
 
Beckman Coulter
•
Assay: Access BNP (Item Number 98200, original formulation)
•
Instrument: Beckman Coulter Access 2 Immunoassay System (P/N 81600N)
Materials required for the comparator test(s) and analyzer model(s) used shall be in accordance 
with the manufacturer instructions for use.
3.6. Intended Use
Reagent
(Item Number 
D06227)
The Access BNP test is intended for use with the Beckman Coulter 
DxI Access Family of Immunoassay Systems for the In Vitro 
quantitative measurement of B-type natriuretic peptide (BNP) in 
plasma specimens using EDTA as the anticoagulant. The test is 
intended to be used for the following indications:
•
as an aid in the diagnosis of congestive heart failure (also 
referred to as heart failure)
•
as an aid in the assessment of severity of congestive heart 
failure 
•
for the risk stratification of patients with acute coronary 
syndromes
•
for the risk stratification of patients with heart failure
Reagent 
(Item number 98200, 
original formulation)
The Access BNP test is intended for use with the Beckman Coulter 
Access Family of Immunoassay Systems for the In Vitro 
quantitative measurement of B-type natriuretic peptide (BNP) in 
plasma specimens using EDTA as the anticoagulant. The test is 
intended to be used for the following indications:
•
as an aid in the diagnosis of congestive heart failure (also 
referred to as heart failure)
•
as an aid in the assessment of severity of congestive heart 
failure
•
for the risk stratification of patients with acute coronary 
syndromes
•
for the risk stratification of patients with heart failure
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 12 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Calibrators
(Item Number 98202)
The Access BNP Calibrators are intended to calibrate the Access 
BNP test for the quantitative determination of BNP levels in human 
EDTA plasma using the Beckman Coulter Access Family of 
Immunoassay Systems.
QC
(Item Number 98201)
The Access BNP QC Controls are intended for monitoring the 
performance of the Access BNP test using the Beckman Coulter 
Access Family of Immunoassay Systems.
3.7. Training and Proficiency
Study Protocol training is conducted by Sponsor with study site personnel prior to site initiation to 
develop familiarity with the investigational assay testing procedure(s) and data export/transfer 
procedure(s). This may also involve performing instrument startup, quality control, sample 
preparation, sample handling and testing. Investigation-use only (IUO) Instructions for Use (IFU) 
documents are to be reviewed and all steps of the study are to be understood prior to the study 
start. 
Sites are to run through all procedural steps to ensure understanding of the study requirements. 
Data collected during familiarization is not used in the study analysis.
Site personnel are expected to have proficiency in clinical laboratory standard operating procedure 
(SOP).
3.8. Investigational Sites
The research is expected to be at a minimum of one (1) and a target of three (3) external testing 
sites.
The participating sites shall have:
•
Ability to perform testing on the Beckman Coulter DxI 9000 Access Immunoassay Analyzer 
(P/N C11137) and Beckman Coulter Access 2 Immunoassay System (P/N 81600N).
•
Appropriately trained staff to perform study testing requirements. 
•
A refrigerator, -20°C and -70°C freezer (or colder) to store reagents, calibrators, QC 
material, and study specimens within the specific temperature requirements of each product 
instruction for use.  
3.9. Study Duration
The study duration is estimated to last approximately 12 months for sample testing. However, study 
sample testing is officially complete when the site completes testing of the sample count provided 
by Sponsor. This may be more or less than the estimated 12 months. Data analysis may take 
another estimated 3 months. 
4. Source Documents and Access to Source Data/Documents
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 13 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Study staff will maintain appropriate medical and research records for this study, in compliance with 
ICH E6, Section 4.9 and regulatory and institutional requirements for the protection of confidentiality 
of subjects.  Study staff will permit authorized representatives of Beckman Coulter, Inc. and 
regulatory agencies to examine (and when required by applicable law, to copy) research records for 
the purposes of quality assurance reviews, audits, and evaluation of the study safety, progress and 
data validity.  
Source documents may include instrument files, maintenance logs, temperature logs, etc. 
Instrument files may be captured as outlines in section 5.2 while paper-based logs may be 
maintained by the site and uploaded electronically. Study staff and sponsors staff will have 
controlled access.
5. Data Handling and Record Keeping
The investigator is responsible for ensuring the accuracy, completeness, legibility, and timeliness of 
the data reported.  All source documents should be completed in a neat, legible manner to ensure 
accurate interpretation of data.  The investigators will maintain adequate instrument data source 
documentation.  
5.1.   Data Management Responsibilities
Data collection and accurate documentation are the responsibility of the study staff under the 
supervision of the investigator.  All source documents and laboratory reports must be reviewed 
by the study team and data entry staff, which will ensure that they are accurate and complete.  
Unanticipated problems must be reviewed by the investigator or designee.  
5.2.  Data Capture Methods
Data is to be directly captured from the laboratory instruments that perform testing. Where 
possible, the site is to electronically transfer laboratory test result instrument data to Beckman 
daily or as agreed upon by Sponsor. Data extraction and transfer instructions are to be provided 
during site initiation training and retained as study-specific documentation in the Investigator 
Site File (ISF). 
An electronic data capture system is not planned for this study. If one is required because the 
testing site cannot provide an output of laboratory test results directly from the instrument, it is to 
be password protected with appropriate data quality checks for an electronic data system. If an 
additional data capture system is required (e.g., eCRF), site personnel shall receive proper 
training coordinated by the Sponsor.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 14 of 38
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Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
5.3.  Types of Data
Data collected is laboratory information on assay test performed on the Beckman Coulter DxI 
9000 Access Immunoassay Analyzer (C11137) and Beckman Coulter Access 2 Immunoassay 
System (P/N 81600N) as required by this protocol.
5.4. Study Records Retention
Study records will be maintained for at least three years post market approval. 
6. Ethics/Protection of Human Subjects
6.1.  Ethical Standard
The investigator will ensure that this study is conducted in full conformity with the principles set 
forth in The Belmont Report: Ethical Principles and Guidelines for the Protection of Human 
Subjects of Research, as drafted by the US National Commission for the Protection of Human 
Subjects of Biomedical and Behavioral Research (April 18, 1979) and codified in 45 CFR Part 
46 and/or the ICH E6. 
6.2. Institutional Review Board/Independent Ethics Committee
The protocol, and other applicable study materials will be submitted to the IRB/IEC for review 
and approval.  Approval of both the study and the protocol must be obtained before site is 
initiated. As applicable, amendments to the protocol will be reviewed for required submission to 
the IRB/IEC for review and approval before the changes are implemented in the study.  
6.3.  Participant Confidentiality
Participant confidentiality is strictly held in trust by the investigators, study staff, and the 
sponsor(s) and their agents.  This confidentiality is extended to cover testing of biological 
samples and genetic tests in addition to any study information relating to participants.
The study protocol, documentation, data, and all other information generated will be held in 
strict confidence.  No information concerning the study, or the data will be released to any 
unauthorized third party without prior written approval of the sponsor. Site personnel do not 
have access to medical standard of care test results or other subject identifying data beyond a 
sample identification (ID) number for the purposes of conducting this protocol required testing 
procedures. Only subjects that consented to this use of the collected samples have been 
included for testing.
The samples used in this testing protocol are collected under protocol BNP-05-24 Access BNP 
Assay Clinical Subject Sample Collection Enrollment Study Protocol. Samples that will be 
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
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Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
included in analysis after testing are from subjects that were appropriately consented and have 
not been withdrawn or discontinued, met study inclusion/exclusion criteria, and provided 
sufficient specimen sample volume required for testing.
Samples collected under protocol BNP-05-24 are anonymized to this protocol’s testing sites and 
in compliance with regulations on data privacy and patient confidentiality. 
7. Objectives
7.1.
Study Objectives
The objective is to validate established diagnostic cutoff (rule-out) and assess HF severity on 
the IUO Access BNP Assay using the DxI 9000 Access Immunoassay Analyzer (P/N C11137). 
A method concordance analysis will be conducted between the IUO Access BNP (item number 
D06227) for DxI 9000 Access Immunoassay Analyzer (P/N C11137) and the Access BNP assay 
(item number 98200; original formulation) for Access 2 Immunoassay System (P/N 81600N).
7.2.
Study Endpoints
Primary endpoints of the study will be diagnostic accuracy between final clinical site diagnosis 
(HF and Non-HF), IUO BNP assay testing results and severity assessment utilizing the NYHA 
class.
8. Study Design 
This is a typical diagnostic device evaluation study with a single arm and no active 
intervention. 
•
Approximately 1100 patient samples (K2 EDTA) collected under BNP-05-24 protocol 
(titled: Access BNP Clinical Enrollment Study) with concentrations that span the 
analytical measuring range will be tested. 
•
Samples are tested with the IUO Access BNP assay (item number D06227) on 
Beckman Coulter DxI 9000 Access Immunoassay Analyzer (P/N C11137) and Access 
BNP assay (item number 98200; original formulation) on Beckman Coulter Access 2 
Immunoassay System (P/N 81600N) one (1) run per day, one (1) replicate per run.
•
Testing will be performed at a minimum of one (1) and approximately three (3) external 
testing sites that have the Beckman Coulter DxI 9000 Access Immunoassay Analyzer 
(P/N C11137) and Access 2 Immunoassay System (P/N 81600N).
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 16 of 38
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Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
•
A minimum of three (3) calibrator lots and a minimum of three (3) IUO reagent 
pack lots (Item number D06227) for DxI 9000 and three (3) IVD reagent pack lots 
(item 98200, original formulation) for Access 2 will be used in this study.
•
Each site will be provided a minimum of one (1) unique calibrator lot and a minimum of 
three (3) individual IUO Access BNP assay reagent pack lots (Item number D06227) for 
DxI 9000 from Sponsor. The IUO reagent lots will be delivered at designated time points 
determined by Sponsor.   
•
Each site will be provided a minimum of one (1) unique calibrator lot and a minimum of 
three (3) individual IVD Access BNP assay reagent pack lots (Item 98200, original 
formulation) for Access 2 from Sponsor. The reagent lots will be delivered at designated 
time points determined by Sponsor.   
9. Study Requirements
9.1. Familiarization Testing
Purpose: Familiarization testing is performed by study personnel prior to starting study testing 
in order to develop familiarization with the investigational assay testing and data submission 
procedures to Sponsor. Results of this testing are used to confirm that quality control 
acceptance criteria are being met for the assay on the Access DxI 9000 Access Immunoassay 
Analyzer at the site. 
Procedure:
•
Familiarization testing includes calibrating the assay and running commercial QC on a 
minimum of one (1) lot of QC as instructed by Sponsor. 
•
Run QC in replicates of two (2) at each QC level each day, over a minimum of three 
(3) days on Beckman Coulter Access DxI 9000 Access Immunoassay Analyzer. 
•
Samples are run on minimum of one (1) calibrator lot and minimum of one (1) reagent lot 
at the site.
•
Verify that Quality Control samples are within the ranges provided by Sponsor.
•
Following completion of the familiarization testing, Sponsor is to review the data and 
notify the laboratory when to proceed with the study testing.
Familiarization results may be used to fulfill the familiarization study requirement in other 
protocol(s) that the site participates in (or vice versa) if the same instrument and personnel are 
used and the familiarization procedure requirements outlined in this protocol are met.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 17 of 38
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Confidential - Company Proprietary
9.2.
Instrument Maintenance and Qualification
If applicable, instrument installation and installation qualification/operational 
qualification/performance qualification (IQ/OQ/PQ) are to be performed for each Beckman 
Coulter DxI 9000 Access Immunoassay Analyzer (P/N C11137). 
Each day prior to study testing, all required routine maintenance must be completed and 
documented according to manufacturer’s required instructions and laboratory policies for all 
instruments and/or equipment utilized in the study. 
Per manufacturing instructions for the Beckman Coulter DxI 9000 Access Immunoassay 
Analyzer (P/N C11137) instruments:
•
The user interface notifies you when maintenance tasks are due, provides instructions for 
performing the maintenance tasks, and contains a maintenance log to record that the 
maintenance was performed.
•
Periodic maintenance includes the following types of maintenance:
o Time-Interval Maintenance—Maintenance that is scheduled to occur after a specified 
period, such as daily, weekly, monthly, or yearly.
o Test-Interval Maintenance—Maintenance that is scheduled to occur after a specified 
number of tests or operations.
•
The system keeps a maintenance log of all completed maintenance tasks. Maintenance 
records may be saved or printed. 
Additional maintenance may be required at the request of Sponsor. As applicable, to help 
maintain instrument performance and reliability, a trained service representative is to coordinate 
periodic maintenance activities for your analyzer in accordance with the terms of your service 
agreement. 
9.3.
Calibration
The commercial Access BNP Calibrators (item number 98202) are provided at six (6) levels: 
zero (0), and approximately 25, 100, 500, 2,500, and 5,000pg/mL. Refer to calibration card for 
exact concentrations.
Assay calibration data are valid up to 28 days. Run the calibrators in duplicate. 
9.4.
Comparator Method
The calibration and maintenance of the Access 2 Immunoassay System (P/N 81600N) should 
be performed per manufacturer’s instructions.
9.5.  Quality Control Analysis
•
Commercial Access BNP QC (item Number 98201) is to be run in replicates of two (2) at 
each QC level each day prior to performing study testing procedure.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
Page 18 of 38
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Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
o
The results of the QC testing are to be evaluated against ranges provided by 
Sponsor. 
 
•
Commercial QC for the Access 2 Immunoassay instrument is to be run in replicates of two 
(2) each day prior to performing study testing procedure.
9.6. Quality Control Acceptance Criteria
•
Site study staff must verify that all control levels meet acceptance criteria before analyzing 
any study samples.  
o
Do not thaw any test samples until results of quality control have met 
specifications.
o
One of the two following scenarios must be true for each level of QC:
o
Both QC replicates are within acceptable ranges provided by sponsor.
o
The mean of two replicates and at least one of the two replicates are 
within the acceptable QC ranges provided by sponsor.
•
If QC acceptance criteria is not met, notify and follow direction of Sponsor. Do not 
proceed to testing study procedure until directed by Sponsor.
o
When outside the acceptable range, the operator is to document any actions taken 
to have QC fall within range.
o
Documented information is to be provided to Sponsor. 
9.7.
 Reagent/Product Handling and Storage
IUO ACCESS BNP for DxI 9000 Reagent Pack (item Number D06227) handling and storage:
Stability 
Unopened at 2 to 10°C 
Up to stated expiration date 
After opening at 2 to 10°C 
28 days  
•
Store upright.
•
Refrigerate at 2°C to 10°C for a minimum of two hours before use on the instrument.
•
Signs of possible deterioration are a broken elastomeric layer on the pack or quality control 
values out of range.
•
If the reagent pack is damaged (e.g., a broken elastomer), discard the pack.
ACCESS BNP for Access 2 Immunoassay System Reagent Pack (Item Number 98200, original 
formulation) handling and storage:
Stability 
Unopened at 2 to 10°C 
Up to stated expiration date 
After opening at 2 to 10°C 
28 days  
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
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Confidential - Company Proprietary
•
Store upright.
•
Refrigerate at 2°C to 10°C for a minimum of two hours before use on the instrument.
•
Signs of possible deterioration are a broken elastomeric layer on the pack or quality control 
values out of range.
•
If the reagent pack is damaged (e.g., a broken elastomer), discard the pack.
•
Discard reagents if any discoloration is observed.
ACCESS BNP Calibrator (Item Number 98202) handling and storage:
•
Provided ready to use.
•
Store upright and freeze at -20°C or colder in a non-defrosting freezer away from the freezer 
door.
•
Stable until the expiration date stated on the label when stored at -20°C or colder.
•
Vial is stable at 2 to 10°C for 30 days after initial use or when removed from frozen storage.
•
Signs of possible deterioration are quality control values out of range.
•
Sponsor will provide exact concentrations.
ACCESS BNP QC (Item Number 98201) handling and storage:
•
Provided ready to use.
•
Store upright and freeze at -20°C or colder.
•
Mix contents thoroughly by gently inverting before use. Avoid bubble formation.  
•
Stable until the expiration date stated on the label when stored at -20°C or colder in a non-
defrosting freezer away from the freezer door.
•
Vial is stable at 2 to 10°C for 30 days after initial use or when removed from frozen storage.
•
Signs of possible deterioration are quality control values out of range.
•
Sponsor will provide QC mean values and standard deviations.
9.8. Sample Handling and Storage
•
Each testing site may receive shipments throughout each month from multiple sample 
collection sites. Upon receipt:
1. Remove temperature data logger (e.g. DeltaTrak®) from shipment container
2. Transfer samples upon receipt to -70 C (or colder) storage. 
3. Follow study guide for temperature data logger (e.g. DeltaTrak®) for data handling 
and upload. 
•
Frozen samples shall be stored frozen at -70°C (or colder) and remain in a frozen state until 
results of quality control specifications are met. 
▪
Record appropriate equipment temperatures on the Temperature Log.
•
Thaw samples only once, allowing to equilibrate to room temperature for a minimum 
of 30 minutes prior to testing. 
o
Complete the Daily Run Log 
o
Do not test samples that meet Exclusion Criteria:
▪
Hemolysis (hemoglobin) sample quality observation 4+ CLSI C56A 
grading scale.6
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
Page 20 of 38
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Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
▪
Icterus (bilirubin) sample quality observation 4+ CLSI C56A grading 
scale.6
•
Testing must be completed on both DxI 9000 and Access 2 instruments within 60 
days of the blood collection date and within 2 hours of removal from the -70 C (or 
colder) freezer on the same aliquot.
1.
After specimen have thawed, they should be mixed by gently inverting several 
times.
2.
Centrifuge following the manufacturer’s instructions for use. 
(Note: Standard recommended instruction is 3,000 RCF for 10 minutes).
3.
Following centrifugation, test specimens following this protocol’s testing 
procedure (See section 11.3 Investigational Method). 
4.
Transfer each sample to appropriate instrument sample cups; follow assay 
requirements for minimum assay dead volumes and assay volume requirements.
•
After testing and valid result confirmed, store aliquots with remaining thawed specimen in 
the -70 C (or colder) freezer.
o
Clearly mark partially used aliquots.
o
Sponsor will give direction on final disposition of the samples prior to the study 
close out visit.
9.9.
Investigational Product Accountability
Sponsor is to document the location of all investigational materials from shipment of
the devices to the testing sites until return or disposal. The testing site investigator is 
responsible for accountability of the investigational device at the site. Access to the 
investigational device/ product is controlled and use is to be limited to the clinical study only by 
authorized staff and in accordance with the protocol. The site is required to maintain the 
inventory of these materials provided and document the reconciliation of these materials. At the 
completion of this clinical study, all unused materials, supplies, and samples and all unused, 
used, expired, or malfunctioning investigational device (reagents or instruments) must be 
returned to Sponsor or disposed of according to instructions from Sponsor. If materials are 
disposed at the study site, the Investigator must provide Sponsor with a signed record of 
disposition. Sponsor may provide a log to the site. The investigator or designee is to keep 
records documenting the receipt, use, return and disposal of the investigational device. The 
records are to include the following (if applicable):
•
Date of receipt
•
Identification of each IVD medical device (e.g., batch number, serial number, or unique 
code)
•
Expiry date
•
Date or dates of use
•
Date on which the investigational device was returned or disposed of, when applicable.
•
Date of return of unused, expired, or malfunctioning product, when applicable.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
Page 21 of 38
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Confidential - Company Proprietary
9.10. Premature Study Termination or Suspension
This study may be suspended or prematurely terminated if there is sufficient reasonable cause.  
Written notification, documenting the reason for study suspension or termination, will be 
provided by the suspending, or terminating party.  If the study is prematurely terminated or 
suspended, the principal investigator will promptly inform the IRB/IEC and will provide the 
reason(s) for suspension or termination.
Circumstances that may warrant termination include, but are not limited to:
•
Insufficient adherence to protocol requirements.
•
Data that are not sufficiently complete and/or evaluable. 
•
Determination of futility.
10.
Study Flow Chart
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
Page 22 of 38
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Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
11. Study Testing
11.1.  Sample/Isolate Requirements
Specimen were collected under protocol BNP-05-24 Access BNP Clinical Subject Sample 
Collection Enrollment Study Protocol from subjects presenting with a clinical suspicion of new 
onset heart failure or worsening symptoms suggestive of decompensated or exacerbated heart 
failure.
•
K2 EDTA plasma samples are collected with IRB/IEC oversight and in accordance with 
local laws and regulations.  
•
Samples are expected to reasonably represent the U.S. demographic population. 
•
Samples shall be preserved in frozen state and stored frozen -70oC (or colder) until ready 
for testing (i.e., results of quality control specifications are met).
•
Sponsor will monitor for sample concentration across the Analytical Measuring Range 
(AMR) with the Access BNP assay on the DxI 9000 Immunoanalyzer (i.e., 5 – 5000 pg/mL).
11.2.
Comparator Method
The calibration and maintenance of the Access 2 Immunoassay System (P/N 81600N) should 
be performed per manufacturer’s instructions.
11.3.
 Investigational Method
•
Each site to receive K2 EDTA subject samples, collected under a separate protocol, for 
testing in approximately monthly shipments, or at other intervals determined by Sponsor, for 
the duration of the study.  
•
Testing must be completed on both the DxI 9000 Access Immunoassay Analyzer (P/N 
C11137) and Access 2 Immunoassay System (P/N 81600N) within 60 days of the blood 
collection date and within 2 hours of removal from the -70 C (or colder) freezer on the 
same aliquot.
•
Refer to section 9.8 Sample Handling and Storage for a detailed instruction.
•
Samples are tested with the IUO Access BNP assay on Beckman Coulter 
DxI 9000 Access Immunoassay Analyzer (P/N C11137) and Access 2 Immunoassay 
System (P/N 81600N).
•
Perform one (1) run, one (1) replicate for each specimen.
o
Confirm test result on DxI 9000 is valid for each study subject (i.e., valid test value 
and no instrument flags). 
o
If testing needs to be repeated on DxI 9000 or Access 2, perform on the same 
sample aliquot within the 2 hours of sample removal from -70 C freezer.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
Page 23 of 38
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Confidential - Company Proprietary
o
If testing needs to be repeated on DxI 9000 AND outside of 2 hours sample 
removal from -70 C freezer, thaw new aliquot to perform testing on BOTH DxI 
9000 and Access 2.  
o
If testing on DxI 9000 is valid, but Access 2 in invalid and outside of 2 hours 
removal from the freezer, then do not thaw a new sample and do not test on 
Access 2. Document appropriately.  
o
NOTE: Site shall electronically transfer all laboratory test data instrument file to 
Sponsor (Beckman) at a minimum daily or as agreed upon by Sponsor.
•
Results shall not be invalidated, eliminated, or discarded by the site.
o
If Sponsor determines the test result(s) cannot be included due to the type of 
protocol deviation, re-testing shall be appropriately documented and repeated as part 
of the corrective action.
12.
Assessment of Safety and Product Quality
To manage health and safety reporting and any product quality issues identified during the study, 
the clinical site is to consult and involve Sponsor. Cross functional teams that may get involved 
include but are not limited to the following: Clinical Affairs, Data Management, Service and Support, 
Supply Chain Management, Complaint Handling, Customer Technical Support, Product 
Development, and Regulatory Affairs. Reporting of issues shall follow internal Sponsor protocols 
and/or standard operating procedure(s). Methods to address product safety and quality issues is be 
documented in a log housed in Sponsor Master File and in the Investigator Site File (ISF). 
The test device is an in vitro diagnostic device that does not come in contact with patients; no 
patient adverse reactions or complications are expected. If the investigational product results in the 
injury, death, or serious medical deterioration of the study staff, or if the potential for death, injury or 
serious medical deterioration exists, the Investigator must immediately contact Sponsor study site 
manager.
In the event the Principal Investigator identifies the test device does not meet safety or product 
quality expectations, they must immediately report findings to Sponsor study site manager.
For any report of product safety and quality issues, a report documented by study site staff is 
submitted to Sponsor study site manager to review and address. 
13.
 Safety Warnings and Precautions
The Access BNP assay is FOR INVESTIGATIONAL USE ONLY on the DxI 9000 Access 
Immunoassay Analyzer (P/N C11137). The performance characteristics of this product have not 
been established.  No clinical decision or patient notification may be made based upon results using 
this investigational use system. 
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
Page 24 of 38
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Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Samples and blood-derived products may be routinely processed with minimum risk using the 
procedure described. However, handle these products as potentially infectious according to 
universal precautions and good clinical laboratory practices, regardless of their origin, treatment, or 
prior certification. Use an appropriate disinfectant for decontamination. Store and dispose of these 
materials and their containers in accordance with local regulations and guidelines.
14.
Clinical Site Monitoring
Clinical site monitoring is conducted to ensure that the rights of human subjects are protected, that 
the study is implemented in accordance with the protocol and/or other operating procedures, and 
that the quality and integrity of study data and data collection methods are maintained.  Monitoring 
for this study will be performed by Beckman Coulter®, Inc. The monitor will evaluate study 
processes and documentation based on the International Conference on Harmonization (ICH), E6: 
Good Clinical Practice guidelines (GCP).
Remote data monitoring is performed on instrument data (including test results, QC, and calibration 
data), temperature records, and service records. Remote data monitoring of instrument data is 
completed weekly at a minimum and approximately monthly for other records.  
15.
Regulatory and Administrative Requirements
15.1. Responsibility of Investigator
The obligations of the Principal Investigator(s) are as detailed in the Confidentiality Agreement, 
Clinical Studies Agreement, and the Investigator Protocol Signature Page.  
In general terms, it is the responsibility of the Principal Investigator(s) to:
•
Ensure that the clinical study is conducted in full conformance with all requirements of the 
clinical study protocol, study agreement, applicable ICH guidelines including Good Clinical 
Practice, 21 CFR 812.43 and other applicable FDA regulations, local and/or institutional 
regulations, and with all requirements of an Institutional Review Board (IRB)/ Independent 
Review Committee (IEC), if applicable.
•
Keep any agreement, contract, or register that stipulates the responsibilities, attributions, 
and functions of all those involved in the clinical performance study.
•
Maintain records of specimen/investigational product accountability and 
specimen/investigational product integrity.
•
Disclose potential conflicts of interest, including financial, that could interfere with the 
conduct of the clinical performance study or interpretation of results as required by 21 CFR 
Part 54.
•
Until either a letter of IRB/IEC approval or a letter stating IRB review and approval are not 
required has been provided to Sponsor.
•
Provide any required updates of study progress to the IRB/IEC.
•
Notify the IRB/IEC when the study is interrupted, discontinued, or completed.
•
Ensure that the study is conducted in accordance with Good Clinical Practice guidelines.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 25 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
•
Comply with all applicable privacy laws, e.g., Health Insurance Portability and Accountability 
Act (HIPAA), Safe Harbor, etc.
•
Ensure that study personnel are experienced with the Reference Method(s) used in this 
study.  
•
Ensure masking of samples, as required.
•
Maintain an adequate internal quality control program and, if requested, to provide the 
sponsor with a copy of the laboratory’s relevant calibration and maintenance procedures.
•
Document relevant study-related communications.
•
Permit the sponsor to monitor and audit study records (including source documents), if 
requested. Document review will be conducted in accordance with institutional requirements 
relating to the protection of study subject confidentiality, when applicable.
•
Permit inspection of relevant study documents by regulatory agency personnel, as 
necessary.
•
Immediately report any protocol deviation to the Sponsor and follow this initial report with 
any required documentation.
•
Ensure study personnel are available during the study to answer questions and to provide 
requested information in a timely manner.  
•
Ensure study records are stored so that they are secure from damage (e.g., protected from 
water damage or fire).  
•
Ensure study records are stored in a way that maintains confidentiality (as specified in the 
Confidentiality Agreement and the Clinical Study Agreement).
•
Ensure that all associates, colleagues, and employees assisting in the conduct of the study 
are informed about their obligations in meeting the above commitments.
Retain the clinical study records until notified by Sponsor that the records may be destroyed or 
returned, postpaid, to Sponsor (refer to Code of Federal Regulations 21 CFR, 812.140, 
Records).  If the Principal Investigator (or designate) needs to change custody of, or change the 
location of the clinical study records, the Principal Investigator (or designate) is responsible for 
contacting Sponsor to work out record retention logistics.
15.2.  Responsibility of Sponsor
It is the responsibility of the Sponsor to:
•
Provide the Investigator with a Clinical Study Protocol and any subsequent amendments.  
•
Provide the Investigator with a Clinical Study Agreement.
•
Provide the Investigator with clinical samples as required.
•
Provide the Investigator with QC values for QC lots.
•
Make payments to the Principal Investigator or to the Investigator’s institution according to 
the Clinical Study Agreement.  
•
Conduct auditing and monitoring of the site; reviewing the monitoring report(s) and following 
up any action(s) required in the monitoring report(s) taking prompt action to secure 
conformity with all clinical performance study requirements.
•
Maintain accountability of IVD medical devices under investigation throughout the clinical 
performance study
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 26 of 38
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Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
•
Document correspondence with all parties involved in the clinical study protocol including, 
when applicable, ethics committees and regulatory authorities
•
Ensure that appropriate mechanisms are in place to cover the liability of the sponsor for the 
study (e.g., insurance)
•
Provide study-specific training to the site study personnel prior to the study’s initiation as 
needed.  
•
Review the clinical study protocol with the Investigator and the study personnel prior to the 
study’s initiation.  
•
Provide the reagents and materials listed in the appropriate section of this clinical study 
protocol.  
•
Maintain the integrity of any study samples that are also required for the standard of care.
•
Take prompt action to secure conformity with all study requirements. 
•
Ensure that IRB/IEC approval or waiver is obtained prior to study initiation at the site. 
•
Provide the Investigator with a final report.
15.3. Protocol Deviations
A protocol deviation is any noncompliance with the clinical study protocol, Good Clinical 
Practice, or Manual of Procedures requirements.  The noncompliance may be on the part of the 
subject, the investigator, or study staff.  
As a result of deviations, corrective actions are to be developed by the study staff and 
implemented promptly. These practices are consistent with investigator and sponsor obligations 
in ICH E6:
•
Compliance with Protocol, Sections 4.5.1, 4.5.2, 4.5.3, and 4.5.4.
•
Quality Assurance and Quality Control, Section 5.1.1
•
Noncompliance, Sections 5.20.1 and 5.20.2.
No testing results shall be invalidated, eliminated, or discarded by the site. All deviations from 
the protocol must be addressed in source documents, reported to sponsor in a timely manner, 
and reported to the local IRB/IEC in accordance with IRB/IEC requirements. It is the 
responsibility of the Principal Investigator to determine if the IRB/IEC should be notified of this 
event. 
15.4.
Study specific or unique noncompliance concerns
Any noncompliance issue impacting the rights, welfare, and safety of human subjects are 
classified as a Major noncompliance and will be escalated following Beckman Coulter’s 
standard operating procedure(s) and or Corrective Action Preventive Action (CAPA) process.
15.5.    Changes to Protocol
In the event the study protocol is amended, Beckman Coulter will provide the Principal 
Investigator with the protocol amendment(s) and a revised protocol.  It is the responsibility of the 
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 27 of 38
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Confidential - Company Proprietary
Principal Investigator to determine if the protocol amendment(s) should be submitted for IRB 
approval.  
Protocol amendments that affect the subject rights, safety, or welfare must be submitted to and 
approved by the IRB.  The sponsor must be notified of the approval, in writing, prior to 
implementation. 
Protocol amendments that are unrelated to subject rights, safety, or welfare may be submitted 
to the IRB at the discretion of the Principal Investigator.  The Principal Investigator or designee 
must notify the sponsor, in writing, when he/she has determined that IRB approval is not 
required, prior to implementation.  
15.6. Reconciliation of Study Material, Supplies and Samples
Sponsor is to supply enough quantities of study materials, supplies, and samples to enable the 
site to complete the study.  The site is required to maintain the inventory of these materials 
provided and document the reconciliation of these materials. 
 
At the completion of this clinical study, all unused materials, supplies, and samples must be 
returned to the sponsor or disposed of according to instructions from the sponsor.  If materials 
are disposed at the study site, the Investigator must provide the sponsor with a signed record of 
disposition.
16.
Statistical Considerations
The number of K2 EDTA individual subject samples used in this testing study will be approximately 
1100. 
All valid testing results of samples collected under protocol BNP-05-24 Access BNP Clinical Subject 
Sample Collection Enrollment Study Protocol from subjects presenting with a clinical suspicion of 
new onset heart failure or worsening symptoms suggestive of decompensated or exacerbated heart 
failure will be included in analysis.
Primary endpoints of the study will be diagnostic accuracy between final clinical site diagnosis (HF 
and Non-HF), IUO BNP assay testing results and severity assessment utilizing the NYHA class.
Method concordance will be established using the valid testing data from the investigational device 
Access BNP Assay (item number D06227) for DxI 9000 Access Immunoassay Analyzer (P/N 
C11137) against the currently marketed Access BNP Assay (item number 98200, original 
formulation) for Access 2 Immunoassay System (P/N 81600N). Positive percent agreement (PPA) 
and Negative percent agreement (NPA) will be calculated using the data. 
The objective is to validate established diagnostic cutoff (rule-out) and assess HF severity on the 
IUO Access BNP Assay (item number D06227) using the DxI 9000 Access Immunoassay Analyzer 
(P/N C11137). Corresponding sensitivity and specificity at this cutoff will be established for the 
intended use population. Diagnostic parameters (i.e., sensitivity, specificity, NPV, and PPV) will be 
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
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Confidential - Company Proprietary
defined by subjects’ HF status as indicated by the IUO assay(s) relative to subjects’ clinical HF 
diagnosis.  Additionally, correlation between Access BNP assay results and severity (NYHA class) 
will be evaluated using the Jonckheere-Terpstra test. Point estimates, event counts, and a 2-sided 
95% score confidence interval will be reported for all relevant endpoints.
In addition to the endpoint results, descriptive statistics such as mean median, standard deviations 
are to be reported. The results are used to support regulatory submissions for the Access BNP 
assay in all geographic regions, including but not limited to the European Union and United States 
(US).
The following requirements are strictly followed to prevent and minimize potential biases:
•
Any DxI 9000 results deemed valid per the instruments’ instruction for use cannot be 
excluded for any reasons other than documented human errors.
•
Access 2 valid result can only be excluded from analysis in the case a retest is needed on 
DxI 9000 and outside of the 2-hour window of sample removal from the -70C (or colder) 
freezer.  In this case a new sample is thawed and tested on both DxI 9000 and Access 2.  
The first Access 2 valid result would be excluded from analysis.  
•
Discrepancy resolution information may be obtained on individual cases for learning 
purpose only. Such information is not to be reported in any forms of estimation or statistics.
17.
Study Materials
The study materials and supplies that will be provided by Beckman Coulter are: 
•
Testing Specimen/Samples
•
IUO Reagent kits for Beckman Coulter DxI 9000 Access Immunoassay Analyzer*
•
IVD Reagent kits for Beckman Coulter Access 2 Immunoassay System
•
IVD Calibrators
•
IVD Quality Controls 
•
Clinical Study Binder – eTMF Florence
•
USB Flash Drives (if applicable)
•
DxI 9000 consumables (as needed)
*  Inventory of Investigational Use Only (IUO) material must be maintained, and reconciliation 
procedures provided by Sponsor after study completion must be followed.
Basic lab equipment and supplies are not provided by sponsor such as centrifuges, pipettors, lab 
coats and gloves. Such supplies are part of site lab operation at the oversight of the Primary 
Investigator.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
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18.
Supplemental Materials Provided 
 
As applicable, these documents may be provided to the investigational site by Sponsor to support 
the conduct of the protocol, however, are not considered part of the protocol. Modifications to these 
documents may be done and do not require an amendment to the protocol.
•
Study Guide (if applicable)
•
IUO Instructions for Use document **
•
Material Safety Data Sheets 
•
Instrument Operator’s Manuals 
•
Biosafety Precautions 
•
Laboratory Handling 
** A change to IUO Instructions for Use document would prompt a protocol review to be conducted 
by Sponsor and if deemed necessary, shall require protocol amendment. 
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
Page 30 of 38
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Confidential - Company Proprietary
19.
Literature References
1.
Mueller T, Gegenhuber A, Poelz W, et al. Diagnostic accuracy of B type natriuretic peptide and 
amino terminal proBNP in the emergency diagnosis of heart failure. Heart 2005; 91:606-612. 
2.
Friedewald V, Burnett J, Januzzi J, Roberts W, et al. The Editor’s Roundtable: B-Type 
Natriuretic Peptide. The American Journal of Cardiology. www.AJConline.org 
3.
Heidenreich PA, Bozkurt B, Aguilar D, et al. 2022 AHA/ACC/HFSA Guideline for the 
Management of Heart Failure: A Report of the American College of Cardiology/American Heart 
Association Joint Committee on Clinical Practice Guidelines. J Am Coll Cardiol 2022;Apr 1.
4.
Piotr Ponikowski, Adriaan A. Voors, Stefan D. Anker, et. al. 2016 ESC Guidelines for the 
diagnosis and treatment of acute and chronic heart failure. Eur J Heart Fail. 2016 
Aug;18(8):891-975. 
5.
Maisel AS, Krishnaswamy P, Nowak RM, McCord J, Hollander JE, Duc P, et al. Breathing Not 
Properly Multinational Study Investigators. Rapid measurement of B-type natriuretic peptide in 
the emergency diagnosis of heart failure. N Engl JMed. 2002; 347:161–167.
6.
2022 AHA/ACC/HFSA Guideline for the Management of Heart Failure: A Report of the 
American College of Cardiology/American Heart Association Joint Committee on Clinical 
Practice Guidelines. J Am Coll Cardiol 2022;Apr 1.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
Page 31 of 38
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APPENDIX A: Investigator Protocol Signature Page
PROTOCOL SIGNATURE PAGE
Protocol:
Access BNP on DxI 9000 Clinical Performance Evaluation: 
Specimen Testing and Clinical Concordance Study
Protocol Number:
 
BNP-02-23
Version:
Site Name:
__________________________________________
The signature below attests that I have read and understand the contents of this protocol (or 
revisions to the protocol) and will adhere to the study protocol requirements as presented 
including all statements regarding confidentiality, and according to local legal and regulatory 
requirements and applicable U.S. Federal Regulations and ICH guidelines.
Investigator Printed Name:
_____________________________________
Investigator Signature:
_____________________________________
Date (dd/mmm/yyyy):
__________________________________
2.7
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
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Testing and Clinical Concordance Study
Protocol Version: 
Page 32 of 38
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Confidential - Company Proprietary
Content Change Abstract
Revision Level
Date
Author Name
Change Description
1.7
1/19/2024
Janicia Harris
Initial Release
1.13
6/18/2024
Polly Robar
• Cover Page: Updated DHF deliverable number 
and sponsor address. Removed “Assay” from 
the document title.
• Updated the Long Title on header and title page 
for purposes of better accuracy. Short title and 
protocol ID remains unchanged.
• List of Abbreviations: Added AMR, CKD, and 
eTMF.
• Throughout document:
-Corrected name of Access 2 from Analyzer to 
Immunoassay System
-Clarified EDTA sample type if K2 EDTA
-Changed source of samples from HF-01-19 to 
BNP-05-24.
-Updated the number of samples to approximately 
1100 per BNP-05-24.
-Updated from Original Formulation to Formulation 
as per K033383.
-Refer to the Access BNP assay as the “currently 
marketed Access BNP” for clarification.
• Section 2 Study Synopsis
-Updated the purpose statement for improved 
clarity of the research purpose to validate a 
comparison of results.
-Study Duration has been extended to 12 
months to accommodate a fresh blood draw 
sample requirement.
-Target Population updated. Samples to be 
tested will now be from a prospective blood draw 
study (BNP-05-24 protocol ID). Samples will 
continue to be frozen in the -70C or colder 
temperature requirement. Testing will be in 
parallel within the sample stability claim of 60 
days (-20C).
-Updated inclusion and exclusion criteria.
No inclusion criteria as samples come from 
BNP-05-24. Exclusion criteria is testing samples 
with HIL observation of 4+ hemolysis or icterus.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 33 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Revision Level
Date
Author Name
Change Description
-Study Design updated to a minimum of three 
(3) calibrator lot and three (3) IUO reagent pack 
lots will be used in this study.
Each site will be provided a minimum of one (1) 
unique calibrator lot and minimum of three (3) 
individual IUO reagent pack lots from Sponsor. 
The IUO reagent lots will be delivered at 
designated time points determined by Sponsor.
• Update to section 6.3 to reflect subjects are 
consented under the BNP-05-24 protocol for 
samples used in this testing study. 
• Update to section 7.2 Study End Points: 
“Results are compared against acceptance 
criteria.”   
• Update to section 8 Study Design - see above 
and section 2 Synopsis.
• Update to section 9.1 For familiarization testing, 
samples are run on a minimum of 1 calibrator lot 
and a minimum of 1 reagent lot at the site.  
• Update to section 9.5 Quality Control Analysis to 
state results of QC will be evaluated against 
ranges provided by sponsor, removed number 
of standard deviations.  
• Update to section 9.6 QC Acceptance Criteria. 
Requirement for both replicates of any QC level 
to fall within ranges provided by sponsor, or the 
mean of 2 replicates must include one replicate 
in range.
• Update to section 9.8 Sample Handling and 
Storage
Replaced section with new text to follow IFU for 
sample thaw at room temp for 30 minutes 
minimum and test within 60 days of collection 
and 2 hours of sample removal from freezer on 
both instruments on the same aliquot.  
Instructions for use of data logger.  
• Update to section 10.0 Study Flow Chart
Replaced flow diagram to reflect updated 
process steps.
• Update to section 11.1 Sample Requirements. 
To align with enrollment protocol BNP-05-24. No 
spiking, diluting, depleting, pooling or contrived 
samples.   
• Update to 11.3 Investigational Method, Method 
Comparison Study Procedure
Replaced text in this section to include samples 
to testing sites at approximately monthly 
intervals, test on both assays on same aliquot 
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 34 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Revision Level
Date
Author Name
Change Description
within 60 days of blood collection and 2 hours 
removal from -70C freezer. If re-test, must be 
within 2 hours of removal from freezer. If outside 
of 2 hours a new aliquot must be tested on both 
assays / instruments.
• Update to section 16 Statistical Considerations. 
Updated protocol from HF-01-19 to BNP-05-24. 
Method concordance is to validate against the 
currently marketed assay.   
• Update to section 17 Study Materials listing 
study materials and supplies that will be 
provided by BEC.
• Update to section 18 Supplemental Materials 
that will no longer be provided by BEC.
• Update to section 19, added sixth literature 
reference.
• Appendix A: Updated protocol name
Version 2.3
8/21/2024
Polly Robar
• Overall study changed from Method Comparison 
to a Clinical Performance Evaluation Testing 
study.
• Updated the Full and Short Title on header and 
title page to reflect the change in study design. 
• Section 2 Study Synopsis
  -Updated the purpose statement to reflect the    
research purpose to perform specimen testing to 
support clinical validation and the IUO Access 
BNP Assay’s regulatory approval on the DxI 9000 
Access Immunoassay Analyzer.
-Updated study objective to is to validate the 
established diagnostic cutoff (rule-out) and 
assess HF severity on the IUO Access BNP 
Assay using the DxI 9000 Access Immunoassay 
Analyzer. 
-Updated Study design to reflect min 1 calibrator 
lot used on this study.
-Site requirements updated to target of 3 sites 
will be used for testing.  
-Update to Statistical Methods to reflect study 
end point is diagnostic accuracy between final 
clinical site diagnosis (HF or Non-HF) and IUO 
BNP assay testing results. 
•
Update to section 3.2 Study purpose, see 
above from study synopsis.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 35 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Revision Level
Date
Author Name
Change Description
•
Section 3.5 Comparator Device Description 
removed as it is no longer needed. 
•
Update to section 3.7 Investigational Sites 
target of 3 sites will be used for testing
•
Update to section 7.1 Study Objectives to 
validate established diagnostic cutoffs (rule-
out cut-off) and ability to assess HF severity. 
•
Update to section 7.2 Study Endpoints   
Removed Method Comparison and Method 
Agreement as they are no longer applicable.  
Changed endpoint to diagnostic accuracy 
between final clinical site diagnosis (HF or 
Non-HF) and IUO BNP assay testing results.  
•
Update to section 8.0 Study Design to remove 
reference to method comparison study and 
related CLSI guideline, reflect the target of 3 
external sites instead of 1, and removed 
reference to testing on the Access 2 
instrument.  Updated to minimum 1 calibration 
lot to be used on this study.
•
Removed section 9.4 Comparator Method for 
calibration and maintenance as it is no longer 
applicable.
•
Updates to section 9.7 Sample Handling and 
Storage to remove references to testing on 
both instruments on the same aliquot and any 
reference to the predicate Access 2 
instrument.  
•
Updated section 10 Study Flow Chart to 
remove reference to testing on Access 2.  
•
Removed section 11.2 Comparator Method 
for calibration and maintenance as it is no 
longer applicable.
•
New section 11.2 Investigational Method 
updated to remove testing on the predicate 
assay. Now will be testing on DxI 9000 only, 
1 run 1 rep...
•
Updated section 12 Assessment of Safety 
and Product Quality with minor text updates.  
•
Updated section 16 Statistical Considerations 
to reflect new approach and analysis for this 
clinical performance testing study.  Primary 
endpoints of the study will be diagnostic 
accuracy between final clinical site diagnosis 
(HF and Non-HF) and IUO BNP assay testing 
results.
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 36 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Revision Level
Date
Author Name
Change Description
•
Updated section 17 Study Materials to 
remove reagent kits for Access 2 as it is no 
longer needed.  
•
Updated Appendix A protocol signature page 
to reflect the new protocol title.  
Polly Robar
• Overall change to add Clinical Concordance 
analysis of DxI 9000 results vs. Access 2.
• Updated the Full and Short Title on header and 
title page to reflect the change in study design. 
• Throughout document added part number for 
DxI 9000 (C11137) and Access 2 (81600N)
• Section 2 Study Synopsis
-Updated Study Purpose to reflect addition of 
clinical concordance analysis vs BNP assay 
testing on Access 2.
-Updated Study Design to add Method 
Concordance
-Updated Statistical Method to include Method 
concordance.
• Updated section 3.2 Study Purpose to add 
clinical concordance.
• Added section 3.5 Comparator Device 
Description for the Access BNP assay and 
Access 2 Immunoassay system
• Updated section 3.8 Investigational site to 
include the ability to test on the Beckman 
Coulter Access 2 Immunoassay System.
• Updated 5.3 Types of Data to include data 
collected from the Beckman Coulter Access 2 
Immunoassay System.
• Updated section 7.1 Study Objectives to include 
Method Concordance. 
• Updated section 7.2 Study Endpoints to include 
Method Concordance.
• Updated section 8 Study Design to include 
testing on Access 2 Immunoassay System.
• Added section 9.4 Comparator Method to note 
calibration and maintenance of Access 2 
instrument should be performed per 
manufacturer’s instructions.
• Updated section 9.5 Quality Control Analysis to 
mention how commercial QC on Access 2 is to 
be run. 
2.7
2.7
2.7
9/12/2024, 5:09:43 PM
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 37 of 38
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Confidential - Company Proprietary
Revision Level
Date
Author Name
Change Description
•  Updated section 9.8 Sample Handling and 
Storage to require that testing be completed on 
both DxI 9000 and Access 2 on the same 
aliquot.
• Updated section 10 Study Flow to include 
testing on Access 2. A valid result on Access 2 
is not required.  
• Added section 11.2 Comparator Method to note 
calibration and maintenance of Access 2 
instrument should be performed per 
manufacturer’s instructions
• Updated 11.3 Investigational Method to mention 
repeat testing on Access 2 is not required if valid 
result on DxI 9000.  
• Updated section 16 Statistical Considerations: -
Any DxI 9000 results deemed valid per the 
instruments’ instruction for use cannot be 
excluded for any reasons other than 
documented human errors.
• Access 2 valid result can only be excluded from 
analysis in the case a retest is needed on DxI 
9000 and outside of the 2-hour window of 
sample removal from the -70C (or colder) 
freezer.  In this case a new sample is thawed 
and tested on both DxI 9000 and Access 2.  The 
first Access 2 valid result would be excluded 
from analysis. 
• Updated section 17 Study Materials to include 
Access 2 materials.
• Updated Appendix A PI Protocol signature page 
with the correct protocol title.  
• Updated Sr Manager approver name
2.7
2.7
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86

Document BNP-02-23
Access BNP Clinical Performance Evaluation: 
Testing and Clinical Concordance Study
Protocol Version: 
Page 38 of 38
Beckman Coulter, Inc. Confidential
Created using GLB-CA-TMP-0033 rev 2.7
Confidential - Company Proprietary
Approvals
The Project Specific TOC defines the approvers for this document (minimum approvers) and is based 
on GLB-QS-PCD-0046.
Role
Name
Title
Development (R&D)
Mo Quin
Staff Development Scientist 
Development (R&D)
Ryan Saffert
Director, Development Science
Quality
Dominique Davidow
Staff Quality Assurance Scientist 
Regulatory
Stephanie Garth
Principal Regulatory Affairs
Regulatory
Neha Desai
Staff Quality and Regulatory Affairs 
Selling and Marketing
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APPROVALS:
APPROVED ON: (mm-dd-yyyy)
2.7
2.7
Robar, Polly
09/11/2024 19:22:15 (UTC)
Davidow, Dominique
09/11/2024 19:27:15 (UTC)
Garth, Stephanie
09/11/2024 19:47:25 (UTC)
Medina, Sompong J
09/11/2024 20:33:32 (UTC)
Quin, Maureen
09/11/2024 20:56:48 (UTC)
Desai, Neha H
09/12/2024 00:40:42 (UTC)
Sanchez, Yolanda
09/12/2024 11:25:29 (UTC)
Sun, Xuan L
09/12/2024 14:01:31 (UTC)
Bettes, Nicole
09/12/2024 14:02:42 (UTC)
Saffert, Ryan
09/12/2024 16:17:34 (UTC)
Object Name: Access BNP Clinical Performance Evaluation: Testing and Clinical Concordance Study
Doc Title: 2.7.2 Clinical Study Protocols and Statistical Plans (External Clinical)
Effective: 09/12/2024
Status: Released
0900324387c96f86