Evaluation of the Performance of the Glycoscore Biomarkers for the Detection of Clinically Significant Prostate Cancer
Conditions: Cancer of Prostate
Sponsor: Medtechtomarket Consulting Ltd

 
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STUDY NUMBER: M2M-GlycoScore-1 
 
EVALUATION OF THE PERFORMANCE OF THE GLYCOSCORE BIOMARKERS FOR 
THE DETECTION OF CLINICALLY SIGNIFICANT PROSTATE CANCER 
 
SPONSORED BY 
 
Medtechtomarket Consulting Ltd 
 Magazine House  
6 Magazine Point  
Riverbank Rd 
Bromborough  
CH62 3JP 
 UK 
 www.medtechtomarket.com. 
 
 
The information in this document is confidential and is the property of Medtechtomarket 
Consulting Ltd.  The information may be provided only to Investigators, sub-Investigators, 
appropriate trial personnel, members of the Institutional Review Board/Independent Ethics 
Committee and personnel of the authorities to whom it is submitted.  This protocol may not be 
photocopied or discussed with third parties not involved in the trial without prior written 
authorisation from Medtechtomarket Consulting Ltd.   
 
 
RESEARCH ETHICS COMMITTEE APPROVAL DATE: 20/09/2024 
 
 
 
 
 
 
 
 
 
 
 
 

 
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PROTOCOL APPROVAL  
 
This protocol has been read and approved by: 
 
Principal Investigator: 
 
Signature: 
 
 
 
Date: 
  
 
 
 
This protocol has been read and approved by: 
 
Sponsor representative: 
 
 
 
Signature: 
 
 
 
 
Date: 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
INVESTIGATOR PROTOCOL APPROVAL PAGE 

 
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EVALUATION OF THE PERFORMANCE OF THE GLYCOSCORE 
BIOMARKERS FOR THE DETECTION OF CLINICALLY SIGNIFICANT 
PROSTATE CANCER 
 
 
I, the undersigned, have read and understood the protocol and am aware of my responsibilities 
as an Investigator. I agree to conduct the study in accordance with this protocol and any 
subsequent amendments, the Declaration of Helsinki and the laws and regulations of the country 
in which the study is being conducted. 
 
Principal Investigator Name: 
Mr Henry Lazarowicz 
 
Investigator Title: 
Consultant Urological Surgeon 
Investigator Address: 
Royal Liverpool University Hospital 
Mount Vernon Street 
Liverpool 
L7 8XP 
 
 
 
 
 
Investigator Signature: 
Date: 
 
 
 
 
 
 
 
 
 
 
 
 

 
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CONTACT NAMES AND ADDRESSES 
 
PRINCIPAL INVESTIGATOR: 
 
Name: 
Mr Henry Lazarowicz 
Address:  
 
Royal Liverpool University Hospital 
Mount Vernon Street 
Liverpool 
L7 8XP 
 
 
Phone: 
0151 706 6807 
Email: 
henry.lazarowicz@liverpoolft.nhs.uk 
 
SPONSOR: 
 
Name of Company: Medtechtomarket Consulting Ltd 
 
Address: 
Magazine House,  
6 Magazine Point,  
Riverbank Rd,  
Bromborough  
CH62 3JP UK 
 
 
 
 
Email:  
Rebecca.newman@medtechtomarket.com  
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
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TABLE OF CONTENTS 
1 PROTOCOL SYNOPSIS .......................................................................................................................8 
2 REVISION HISTORY .......................................................................................................................... 12 
3 BACKGROUND .................................................................................................................................. 13 
3.1 PSA ............................................................................................................................................. 14 
3.2 MPMRI......................................................................................................................................... 14 
2.3 BIOPSY ......................................................................................................................................... 14 
4 STUDY OBJECTIVES ........................................................................................................................ 15 
3.1 PRIMARY OBJECTIVE ................................................................................................................. 15 
3.2 SECONDARY OBJECTIVES ........................................................................................................ 15 
3.3 PRIMARY ENDPOINT .................................................................................................................. 15 
3.4 SECONDARY ENDPOINTS ......................................................................................................... 15 
3.5 STUDY RISKS AND BENEFITS ................................................................................................... 16 
3.5.1 RISKS ................................................................................................................................. 16 
3.5.2 BENEFITS .......................................................................................................................... 16 
5 CLINICAL STUDY DESIGN ................................................................................................................ 16 
5.1 STUDY PURPOSE ...................................................................................................................... 16 
5.2 STUDY DESIGN ......................................................................................................................... 17 
5.3 JUSTIFICATION FOR SPECIMEN COLLECTION ..................................................................... 17 
5.4 METHODS TO AVOID BIAS ....................................................................................................... 18 
6 DIAGNOSTIC METHOD ..................................................................................................................... 18 
6.1 THE GLYCOSCORE PROSTATE CANCER BIOMARKERS ..................................................... 18 
6.2 DIAGNOSTIC METHODS USED FOR COMPARISON ............................................................. 18 
7 SPECIMENS AND SUBJECTS PROVIDING SPECIMENS ............................................................... 18 
7.1 PATIENT PARTICIPANTS .......................................................................................................... 18 
7.1.1 PATIENT INCLUSION CRITERIA ...................................................................................... 18 
7.1.2 PATIENT EXCLUSION CRITERIA ..................................................................................... 19 
7.2 SPECIMEN COLLECTION.......................................................................................................... 19 
7.3 EXPECTED DURATION OF SUBJECT PARTICIPATION ......................................................... 19 
7.4 EXPECTED DURATION OF STUDY .......................................................................................... 19 
7.5 END OF STUDY .......................................................................................................................... 19 
7.6 SPECIMEN HANDLING AND PROCESSING ............................................................................ 19 
7.6.1 GLYCOSCORE STUDY BLOOD SAMPLES (ONE EDTA TUBE COLLECTED FOR 
MEDTECHTOMARKET) ............................................................................................................. 20 
7.6.2 LIVERPOOL UNIVERSITY BIOBANK URINE SAMPLE (STORAGE AT LUB FOR FUTURE 
RESEARCH) ............................................................................................................................... 20 
7.6.3 LIVERPOOL UNIVERSITY BIOBANK BLOOD SAMPLE (ONE EDTA TUBE STORAGE AT 
LUB FOR FUTURE RESEARCH) ............................................................................................... 20 
7.7 WITHDRAWAL OF PARTICIPANTS .......................................................................................... 21 
8 STUDY OPERATOR SELECTION AND TRAINING .......................................................................... 21 

 
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9 STUDY PROCEDURES ..................................................................................................................... 21 
9.1 PARTICIPANT RECRUITMENT ................................................................................................. 21 
9.2 SAMPLE COLLECTION .............................................................................................................. 22 
9.3 QUALITY CONTROL PROCEDURES ........................................................................................ 22 
9.4 PSA TEST QUALITY CONTROL PROCEDURES ..................................................................... 22 
10 STUDY MONITORING ....................................................................................................................... 22 
10.1 SOURCE DATA VERIFICATION ................................................................................................ 23 
10.2 DEFINITION OF SOURCE DATA ............................................................................................... 23 
10.3 AUDITING ................................................................................................................................... 23 
11 DATA MANAGEMENT ........................................................................................................................ 23 
11.1 DATA HANDLING ....................................................................................................................... 23 
11.2 CASE REPORT FORM ............................................................................................................... 23 
11.3 RECORD KEEPING .................................................................................................................... 24 
12 STATISTICAL CONSIDERATIONS .................................................................................................... 24 
12.1 DATA QUALITY CONTROL ........................................................................................................ 24 
12.2 STATISTICAL ANALYSIS OF PRIMARY OBJECTIVE .............................................................. 24 
12.3 STATISTICAL ANALYSIS OF SECONDARY OBJECTIVES ..................................................... 25 
12.4 DETERMINATION OF SAMPLE SIZE ........................................................................................ 25 
12.4.1 
SAMPLE SIZE CALCULATIONS ................................................................................ 25 
12.5 PROCEDURE FOR ACCOUNTING FOR MISSING, UNUSED OR SPURIOUS DATA ............ 26 
13 STUDY PROTOCOL AMENDMENTS ................................................................................................ 26 
14 DEVIATIONS FROM THE CLINICAL STUDY PROTOCOL............................................................... 26 
15 STATEMENTS OF CONFORMITY ..................................................................................................... 27 
16 INSURANCE ....................................................................................................................................... 27 
17 ADVERSE EVENTS ........................................................................................................................... 27 
17.1 ADVERSE EVENT DEFINITION AND CATEGORISATION ....................................................... 27 
17.2 REVIEWING AND REPORTING ADVERSE EVENTS ............................................................... 28 
18 INFORMED CONSENT PROCESS.................................................................................................... 28 
18.1 VULNERABLE POPULATION - PATIENTS LACKING CAPACITY TO CONSENT................... 28 
19 SUSPENSION OR PREMATURE TERMINATION OF THE STUDY ................................................. 28 
20 PUBLICATION AND COMMUNICATION POLICY ............................................................................. 29 
21 REFERENCES.................................................................................................................................... 29 
22 APPENDICES ..................................................................................................................................... 30 
22.1 APPENDIX A – STUDY PROCEDURES FLOWCHART ............................................................ 30 
22.2 APPENDIX B – SCHEDULE OF EVENTS .................................................................................. 31 
 
 
 
 

 
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LIST OF ABBREVIATIONS/GLOSSARY 
 
 
 
 
 
 
PSA 
Prostate specific antigen 
PCa 
Prostate cancer 
mpMRI 
Multi-parametric magnetic resonance imaging 
CPG 
Cambridge Prognostic Group 
DRE 
Digital rectal examination 
TRUS 
Transrectal ultrasound guided biopsy 
RUA 
Research Use Only 
ELISA 
Enzyme linked immunosorbent assay 
BPH 
Benign prostatic hyperplasia 
CRF 
Case Report Form 
GCP 
Good Clinical Practice 
ICH 
International Conference of Harmonisation 
AE 
Adverse Event 
SAE 
Serious Adverse Event 
SDV 
Source Data Verification 
PPA 
Positive Percent Agreement 
NPA 
Negative Percent Agreement 
ADT 
Androgen Deprivation Therapy 
LUB 
Liverpool University Biobank 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
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1 
 PROTOCOL SYNOPSIS 
STUDY TITLE 
Evaluation of the performance of the GlycoScore biomarkers for 
the detection of clinically significant prostate cancer 
PURPOSE OF THE 
STUDY  
 
GlycoScore Dx Limited, a UK-based diagnostics company have 
identified three markers, ST6GAL1, GCNT1 & GALNT7, that 
showed promise as biomarkers of prostate cancer in initial marker 
validation studies. The purpose of this study is to further evaluate 
the sensitivity and specificity of the GlycoScore biomarkers for the 
detection of clinically significant prostate cancer. 
Sensitivity and specificity will be determined for each marker, 
combinations of the three markers and combinations of the 
GlycoScore biomarkers with PSA. The results from this study will 
be used to identify the most suitable biomarker/biomarkers for use 
in the GlycoScore test.  
 
SPONSOR 
 
Medtechtomarket Consulting Ltd 
Magazine House  
6 Magazine Point  
Riverbank Rd 
Bromborough  
CH62 3JP 
UK 
 
STUDY DESIGN 
 
This is a prospective, non-interventional study using venous blood 
samples taken from patients with suspected prostate cancer or on 
active surveillance, attending the hospital Urology department for a 
transperineal biopsy of prostate.  
 
Following informed consent, venous blood samples from at least 
137 patients will be collected and sent to Medtechtomarket 
Laboratories for testing. The concentrations of ST6GAL1, GCNT1, 
GALNT7 in each sample will be measured by ELISA assays, and 
the sample PSA will be measured using a CE-marked PSA assay. 
Results from testing of the samples will not be used in the clinical 
management of enrolled patients. 
 
The Investigator will provide Medtechtomarket with all prostate 
cancer related results from each participant including PSA, mpMRI 
(Likert or PI-RADS® score) and biopsy result (if carried out).   
Where prostate cancer is diagnosed, an indication of severity 
(according to the hospital’s preferred method) such as Gleason 
score or CPG classification1 will also be provided. 
 
As part of the consenting process, patients will be asked 
permission for two additional blood samples to be taken at the 
same time as the study sample, alongside a urine sample which 

 
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will be transferred to the Liverpool University Biobank to be frozen 
and archived for use in future prostate cancer research. Patients’ 
will also be asked permission for utilisation of archived biopsy 
tissue samples for potential future research studies. 
 
The Urology Department at Liverpool University Hospitals NHS 
Foundation Trust has a track record of clinical and translational 
research, including studies that have looked at detecting prostate 
and bladder cancer using patient’s urine. These additional samples 
would provide an opportunity for further validation samples that 
would complement the current work and would provide a valuable 
bio-resource for future research studies, both in-house and in 
collaboration with other centres. 
STUDY PRIMARY 
OBJECTIVES 
To evaluate the diagnostic performance of the GlycoScore 
biomarkers for detection of clinically significant prostate cancer 
(Gleason score of ≥7)2.  
SECONDARY 
OBJECTIVES 
• To establish an algorithm for calculation of a ‘GlycoScore’ that 
converts the assay results from the optimal combination of 
biomarkers into a score that indicates the likelihood that a 
patient has clinically significant prostate cancer  
• To evaluate the ability of the ‘GlycoScore’ to distinguish 
between individual Gleason scores and Gleason grade groups 
• To evaluate the ability of the ‘GlycoScore’ to distinguish 
between CPG groups 1 to 5 (Cambridge Prognostic Group 
classification) 
• To compare the ability of the GlycoScore biomarkers to detect 
clinically significant prostate cancer (with and without PSA) to 
PSA alone 
PRIMARY 
ENDPOINT 
The primary endpoint will be a comparison of calculated sensitivity, 
specificity and positive and negative predictive values for detection 
of clinically significant prostate cancer for the following: 
• 
Each individual GlycoScore biomarker, alone and in 
combination with PSA (measured at Medtechtechtomarket 
using a CE-marked assay) 
• 
Each possible combination of GlycoScore biomarkers, 
alone and in combination with PSA (measured at 
Medtechtechtomarket using a CE-marked assay) 

 
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The reference method for diagnosis of prostate cancer will be 
biopsy, and the reference method for identifying patients who do 
not have cancer will be by a combination of mpMRI and/or biopsy.  
SECONDARY 
ENDPOINT 
Secondary endpoints will be: 
• Establishment of an updated ‘GlycoScore’ algorithm 
• Assessment of the correlation between the ‘GlycoScore’ for 
each sample and Gleason score at biopsy 
• Assessment of the correlation between the ‘GlycoScore’ and 
the Cambridge Prognostic Group classification 
• A comparison of the diagnostic performance of the 
GlycoScore biomarkers (with and without PSA) for the 
detection of clinically significant prostate cancer against the 
PSA test (hospital PSA result) alone 
 
SAMPLE SIZE  
A minimum of 137 male adult patients aged 18 and over.  
SUBJECT 
POPULATION 
 
The subject population will consist of male patients who are 
attending the Urology Department for a transperineal prostate 
biopsy for suspected prostate cancer or are on active surveillance 
(patients diagnosed with low-grade, slow-growing localised 
prostate cancer). 
 
Patient Group 
 
Inclusion Criteria 
• Male patients aged 18 years and over 
• The patient is being investigated for suspected prostate 
cancer or is on active surveillance 
• The patient is able to give informed consent to take part in the 
study 
 
Exclusion Criteria 
• A patient who has already taken part in the study 
• A patient with an active urinary tract infection 
• The patient has a prior diagnosis of any other cancer or 
receiving any cancer treatment (including ADT) 
• The patient is currently or within the last 4 months enrolled on 
another study involving an investigational medicinal product. 
 
 
 
STUDY DURATION 
 
The study will run for 12 months.  The maximum duration of 
subject participation in this study is 1 hospital visit (informed 

 
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consent, blood and urine sample collection will be carried out 
during the same visit). 
 
STATISTICAL 
METHODS AND 
ANALYSIS 
SUMMARY 
Data from the study will be analysed using the results of prostate 
biopsy or a combination of mpMRI and biopsy as the reference 
method for diagnosis of PCa. 
ROC curves will be plotted for: 
• 
Each individual GlycoScore biomarker, alone and in 
combination with PSA (measured at Medtechtomarket)  
• 
Each possible combination of GlycoScore biomarkers, 
alone and in combination with PSA (measured at 
Medtechtomarket) 
The ROC curves will be used to calculate sensitivity, specificity, 
likelihood ratio, and positive and negative predictive values for 
that method.  This will also be repeated using the most recent 
PSA result provided by the hospital to compare the diagnostic 
performance of the current PSA test method with the GlycoScore 
biomarkers. 
Suitable correlation analysis will be used to evaluate the 
agreement between the ‘GlycoScore’ and  
• Gleason score & Gleason Grade Group 
• Cambridge Prognostic Group score 
FOLLOW-UP 
EVALUATIONS 
No planned follow up evaluations required. 

 
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2 
REVISION HISTORY  
Amendment 
No. 
Protocol 
Version No. 
Date issued 
Author(s) of 
changes 
Details of Changes made 
N/A 
1.0 
18/06/2024 
N/A 
Original Version submitted to 
IRAS 
1 
2.0 
24/07/2024 
Lindsay 
Traynor 
Addition of ‘End of Study’ 
definition.  
Update to clarify that potential 
participants may be given the 
patient information sheet prior to 
attending the consent and 
donation visit. 
Updated specimen handling 
section to include processing of 
blood samples by LUB (also 
added to glossary) 
Study Procedures Flowchart 
updated. 
Clarification of participant 
information collected following 
consent. 
2 
3.0 
13/09/2024 
Rebecca 
Newman 
Added consent for utilisation of 
archived tissue biopsy samples 
for future research purposes 
3 
4.0 
20/09/2024 
Lindsay 
Traynor 
Added additional Paxgene tube 
to be collected for LUB.  In 
addition to the trust, LUB to hold 
patient identifiable details for 
biobanking purposes only. 
4 
5.0 
23/12/2024 
Lindsay 
Traynor 
Correction to state urine to be 
aliquoted by LUB and 
clarification of LUB process and 
collection kit 

 
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3 
BACKGROUND  
Prostate cancer (PCa) is the most common cancer in men in the UK and the second most 
common cancer in men worldwide. Affecting one in every eight men, more people live with a 
diagnosis of prostate cancer than any other type of cancer. It develops slowly and can remain 
undetected for many years. However, in some cases the cancer may be aggressive and, if 
undetected and unchecked, may spread to other areas of the body. 
 
An urgent and unmet clinical need exists to develop more accurate diagnostic tests which can be 
applied at an earlier stage in the clinical pathway, and can help clinicians differentiate between 
clinically significant prostate cancers and slow-growing cancers that may never cause problems 
or shorten life. 
 
Prostate-specific antigen (PSA) is a protein secreted by the prostate gland that can be detected 
in a patient’s blood and can be used to help identify patients likely to have prostate cancer.  
 
After presenting either with lower urinary tract symptoms or requesting screening for prostate 
cancer, patients typically undergo a digital rectal exam (DRE) and a PSA blood test.  The current 
recommended diagnosis pathway for prostate cancer is outlined in Fig.1. 
 
Fig. 1 NICE diagnostic pathway for suspected prostate cancer,3 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
*Not offered to patients who are unsuitable for radical treatment. 
Symptoms/DRE/PSA 
Urology referral 
*multiparametric MRI as the first-line investigation for 
people with suspected clinically localised prostate 
cancer. Report the results using Likert/PI-RADS® scale  
multiparametric MRI-influenced prostate biopsy 
recommended if Likert or 
 PI-RADS® score is ≥3. 

 
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3.1 PSA 
One of the best-known biomarkers in cancer biology is PSA4,5.  It is a proteolytic glycoprotein 
enzyme belonging to the kallikrein family of serine proteases produced by normal prostate 
epithelium.  PSA is organ-specific but not disease-specific and is detected in both normal 
prostate and benign prostatic hyperplasia, as well as primary and metastatic prostate cancer 
cells meaning levels can be elevated for reasons other than cancer (leading to false positive 
results).  
 
Two thirds of men with a raised PSA level will not have prostate cancer, meaning many men 
undergo unnecessary invasive investigations including prostate biopsy. 
 
Similarly, 1 in 7 men with prostate cancer do not have elevated PSA levels, so a significant 
number of patients with prostate cancer are not diagnosed by PSA also (leading to false negative 
results). 
 
Thus, PSA alone has a poor predictive value for diagnosing or excluding clinically significant 
prostate cancer.  
 
3.2 mpMRI  
Multi-parametric magnetic resonance imaging (mpMRI) creates detailed images of the prostate 
that enables the clinician to better detect suspected prostate cancer.  mpMRI combines the 
morphological assessment of T2-weighted imaging (T2WI) with diffusion-weighted imaging 
(DWI), dynamic contrast-enhanced (DCE) perfusion imaging and spectroscopic imaging (MRSI), 
which prior to biopsy can improve the detection rate of clinically significant PCa by allowing 
targeted samples to be taken. MpMRI can provide valuable clues about the histopathological 
aggressiveness of a PCa lesion and tumour stage.  However, approximately 40% of patients 
have an inconclusive mpMRI result, which can lead to further examinations and repeat biopsies, 
with the associated anxiety, discomfort, risk and cost for both the patient and the NHS6.  
 
 
2.3 BIOPSY 
Prostate biopsies provide a definitive method of confirming prostate cancer, and determining its 
severity by taking samples of tissue from the prostate gland, either by transperineal biopsy (TPB) 
or transrectal ultrasound-guided biopsy (TRUS). 
These samples are collected and analysed by a pathologist who assigns a Gleason Score to the 
detected prostate cancer, which is based upon its morphological appearances under a 
microscope. The Gleason score helps indicate the clinical significance of the prostate cancer, 
with high-grade (potentially aggressive) prostate cancers having Gleason scores greater than 7. 
Prostate biopsy however, is an invasive and uncomfortable procedure associated with significant 
risks of bleeding, infection, sepsis and occasional death. Of those patients undergoing prostate 
biopsy, a significant proportion will have a negative result, meaning they have undergone an 
unnecessary procedure and incurred risks and costs to the NHS that could have been avoided. 
The availability of PCa-specific biomarkers could potentially facilitate a more accurate diagnosis, 
prognostication, risk stratification for treatment and reduce the need for unnecessary biopsies 
and their associated risks. 

 
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4 
STUDY OBJECTIVES  
3.1 PRIMARY OBJECTIVE 
The primary objective of this study is to evaluate the diagnostic performance of the GlycoScore 
biomarkers for clinically significant prostate cancer in blood samples taken from 137 male 
participants with suspected prostate cancer or on active surveillance. In this context, clinical 
significance would be taken as a Gleason score of ≥72.  
 
3.2 SECONDARY OBJECTIVES 
The secondary objectives of the study are: 
• 
To establish an algorithm for calculating a ‘GlycoScore’ that converts the assay results 
from the optimal combination of biomarkers into a score that indicates the likelihood that a 
patient has clinically significant prostate cancer) 
• 
To evaluate the ability of the ‘GlycoScore’ to distinguish between individual Gleason 
scores and Gleason grade groups 
• 
To evaluate the ability of the ‘GlycoScore’ to distinguish between CPG groups 1 to 5 
(Cambridge Prognostic Group classification)1 
• 
To evaluate the ability of the GlycoScore biomarkers (with and without PSA) to detect 
clinically significant PCa against that of PSA alone  
 
3.3 PRIMARY ENDPOINT 
The primary endpoint will be a comparison of calculated sensitivity, specificity and positive and 
negative predictive values for clinically significant prostate cancer for the following: 
 
• 
Each individual GlycoScore biomarker, alone and in combination with PSA 
• 
Each possible combination of GlycoScore biomarkers, alone and in combination with 
PSA 
 
The reference method for diagnosis of prostate cancer will be biopsy, and the reference method 
for identifying patients who do not have cancer will be by a combination of mpMRI and/or biopsy. 
 
 
3.4 SECONDARY ENDPOINTS 
Secondary endpoints will be the following: 
• 
Assessment of the correlation between the ‘GlycoScore’ and the Cambridge Prognostic 
Group classification 
• 
Comparison of the sensitivity, specificity and positive and negative predictive value of 
the GlycoScore biomarkers (with and without PSA measured at Medtechtomarket) 
against that of PSA alone 
• 
Establishment of an updated ‘GlycoScore’ algorithm 
• 
Assessment of the correlation between the ‘GlycoScore’ for each sample and Gleason 
score and Gleason Grade Group at biopsy 

 
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The reference method for diagnosis of prostate cancer will be biopsy, and the reference method 
for identifying patients who do not have cancer will be mpMRI or biopsy. 
 
3.5 STUDY RISKS AND BENEFITS  
3.5.1 Risks 
No anticipated adverse device effects related to this study have been identified. 
As this is an observational study, participant risks are limited to those relating to the 
collection of the blood samples. Participants in this study will be subject to risks that are no 
greater than, or are equivalent to the risks associated with undergoing a peripheral venous 
blood test.  Anticipated whole blood volume sampling should be ≤22.5ml (2 x 10ml and 1 x 
2.5ml tubes).  The use of standard precautions will minimise risk. Since blood sampling is a 
normal part of routine patient care, participating in this study should not cause any additional 
emotional stress or cause additional risks. 
 
Possible side effects from blood drawing include faintness, inflammation of the vein, pain, 
bruising, haematoma or bleeding at the site of the puncture. A peripheral site infection may 
occur in rare circumstances.  
 
Risks to study staff are those relating to contact with blood products. The use of standard 
precautions will minimise this risk. 
 
Urine collection is a routine part of the procedure prior to the transperineal biopsy and 
providing a urine sample for use in future ethically approved research projects should result 
in no additional risk to the participant.  Collection of biopsy tissue samples is the purpose of 
the transperineal biopsy, no additional tissue samples will be required therefore there should 
be no additional risk to the participant. There are no other residual risks associated with the 
study.  
 
3.5.2 Benefits 
The clinical benefit of the GlycoScore Prostate Cancer Test is being able to detect prostate 
cancer and to be able to distinguish between clinically significant and non-significant prostate 
cancer using a blood sample. This could reduce the number of unnecessary biopsies carried 
out and allow faster diagnosis of prostate cancer than the current clinical pathway. There are 
no direct benefits to patients participating in the study and results from the GlycoScore Test 
will not be used to influence patient management or decision making in any way. However, if 
the study leads to the clinical availability of the GlycoScore Prostate Cancer Test, then there 
may be a potential significant benefit to patients in the future.  
 
For patients that agree for their samples to be used in future research, the storage of urine, 
blood and use of archived tissue samples would provide the hosting trust a valuable clinical 
resource for future ethically approved clinical research and material for validating current 
research studies. 
 
5 
CLINICAL STUDY DESIGN 
5.1 STUDY PURPOSE 
Initial studies carried out by Newcastle University using RUO ELISA kits showed significant 
correlation between PCa and the three biomarkers ST6GAL1, GCNT1 and GALNT7.  A further 
cohort of plasma samples from a range of low and high Gleason grade samples as well as BPH 

 
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(benign prostatic hyperplasia) and healthy patients (supplied by York biobank) was used to 
generate a ‘GlycoScore’ algorithm7.   
The purpose of this study is to evaluate the three biomarkers ST6GAL1, GCNT1 and GALNT7 
for their ability to identify clinically significant prostate cancer, either alone, as a combination of 
the biomarkers or in combination with the PSA (measurement obtained at Medtechtomarket). 
ST6GAL1, GCNT1 and GALNT7 concentrations from each plasma sample will be measured 
using the GlycoScore ELISA assay developed by Medtechtomarket.    
The diagnostic presence of prostate cancer will be determined by comparison with biopsy 
results, and controls determined by a combination of mpMRI and negative biopsy results. 
Assessment of the correlation between GlycoScore results with the full range of biopsy Gleason 
scores/Gleason Grade group/CPG grade will allow an assessment of the prognostic value of the 
GlycoScore test and its potential use in the clinical management of prostate cancer. 
 
5.2 STUDY DESIGN 
This is a prospective, non-interventional study using venous blood samples taken from patients 
with suspected prostate cancer or on active surveillance. A venous blood sample for use in the 
study will be obtained from a minimum of 137 patients referred to the Urology Clinic for 
transperineal biopsy for the investigation of suspected PCa or patients undergoing active 
surveillance.  The venous blood and urine samples will be taken during the patient’s routine 
hospital visit, prior to the biopsy procedure taking place.  The blood samples will be collected 
according to the study protocol and sent to Medtechtomarket Laboratories where they will be 
tested using the GlycoScore ELISA assays for all three biomarkers and a PSA measurement 
recorded using a CE marked PSA test.   
 
All blood and urine samples will be transferred to Liverpool University Biobank for processing. 
The GlycoScore study samples will then be transferred to Medtechtomarket for testing and the 
additional blood and urine samples will be frozen and archived at LUB.  Tissue samples will be 
stored in the pathology lab as standard.  
 
The following test results will be supplied by the hospital for each study participant: 
 
• 
Diagnosis or otherwise of PCa           
• 
Hospital PSA test result and date 
• 
mpMRI report including Likert or PI-RADS® score 
• 
Gleason score 
• 
Gleason grade group 
• 
CPG group (if available) 
 
Results obtained at Medtechtomarket from testing of the samples will not be used in the clinical 
management of enrolled study participants. 
 
5.3 JUSTIFICATION FOR SPECIMEN COLLECTION 
The ELISAs used to measure the ST6GAL1, GCNT1 and GALNT7 biomarker concentrations 
require a venous whole blood sample, and therefore whole blood samples are essential for the 
completion of this study. Whenever possible this will be obtained if any blood is required as part 
of the patient's journey.  In cases where blood samples have already been taken prior to study 
recruitment, a separate venous blood sample will be required for the purpose of the study. 

 
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5.4 METHODS TO AVOID BIAS 
All patients referred to secondary care with suspected PCa or on active surveillance and meeting 
the study criteria will be invited to join the study.  No further selection or grouping of participants 
will take place during the study. 
 
Samples and clinical data will be pseudonymised, GlycoScore results from each participant will 
be blinded to the clinician managing their healthcare, so as not to influence their diagnosis and/or 
treatment. 
 
There is unlikely to be any operator bias when performing the GlycoScore assays as each test 
incorporates a calibration curve.  However, a minimum of 2 study staff (laboratory staff) will be 
fully trained to run the GlycoScore immunoassays to minimise further any potential risk of 
Operator bias. All Medtechtomarket study staff will be blinded to patient diagnosis until after the 
samples have been analysed for biomarker concentration. 
 
6 
DIAGNOSTIC METHOD  
6.1 THE GLYCOSCORE PROSTATE CANCER BIOMARKERS   
The aim of the study is to identify whether the three prostate cancer markers identified by 
Glycoscore can be used individually, or in combination, to diagnose significant prostate cancer 
(The GlycoScore Prostate Cancer biomarkers will be measured using quantitative 
immunoassays, with labelled antibodies against the biomarkers ST6GAL1, GCNT1 & GALNT7).  
The immunoassays will measure and report the concentration of the selected biomarkers (or 
combination of) in plasma. Once the most suitable biomarkers have been identified the aim will 
be to generate an algorithm that will convert the assay results into a ‘GlycoScore’ which can be 
used to give an indication of disease severity.   
 
The GlycoScore biomarkers are intended to be used for the detection of clinically significant 
prostate cancer and could potentially be used to distinguish between different grades of PCa. 
The test is intended to be used by trained professionals in a pathology laboratory setting.  
 
6.2 DIAGNOSTIC METHODS USED FOR COMPARISON 
The methods used for comparison in this study will be mpMRI and biopsy. 
 
7 
SPECIMENS AND SUBJECTS PROVIDING SPECIMENS 
7.1 PATIENT PARTICIPANTS 
A minimum of 137 adult male patients will be recruited for the study. 
7.1.1 Patient Inclusion Criteria 
A patient is considered eligible for participation in this part of the study if all the following 
inclusion criteria are satisfied before enrolment: 
 
• Male patients aged 18 years and over 
• Patient is being investigated for suspected prostate cancer or is on active surveillance 
• The patient is able to give consent to take part in the study 
 
 

 
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7.1.2 
Patient Exclusion Criteria 
A patient is considered ineligible for participation in this part of the trial if any  
           of the following exclusion criteria are met prior to enrolment: 
 
• A patient who has already taken part in the study 
• A patient with an active urinary tract infection 
• The patient has a prior diagnosis of any other cancer or is receiving any cancer treatment 
(including ADT) 
• The patient is currently or within the last 4 months enrolled on another study involving an 
investigational medicinal product. 
 
7.2 SPECIMEN COLLECTION  
All specimen collection will be carried out by trained study staff using standard site procedures 
(Appendix A – Study Procedures Flowchart).  Any study blood (and urine) samples must be 
drawn/taken prior to any invasive procedure, such as prostate biopsy.  
 
Two (≤10ml) K2 EDTA and one (≤2.5ml) Paxgene tube of venous blood will be taken on the 
patient’s 1st study visit (this will be during their pre-biopsy check prior to their biopsy); one EDTA 
tube for the study and the other EDTA and Paxgene tube for the University of Liverpool Biobank.  
Study samples will only be collected on a week day. 
 
The 20ml sample of urine obtained from clinic, pre-biopsy and pre-dipstick testing will be 
decanted then transferred, processed and stored at -80°C at the University of Liverpool Biobank. 
 
Following routine processing biopsy tissue samples will be stored in the pathology lab as 
standard practice.  
 
7.3 EXPECTED DURATION OF SUBJECT PARTICIPATION  
The maximum duration of subject participation in this trial is one hospital visit, with screening, 
consent and sample collection taking place on the same visit.  Potential participants may be 
given the patient information sheet prior to attending the consent and donation visit. 
 
7.4 EXPECTED DURATION OF STUDY  
The study is expected to take approximately 12 months to fulfil recruitment targets. 
 
7.5 END OF STUDY  
The end of the study, will be the date that the associated clinical data is collected from the last 
participant recruited onto the trial. 
 
7.6 SPECIMEN HANDLING AND PROCESSING 
Blood samples will be handled, transported and disposed of according to site-specific guidelines.  
Liverpool University Biobank will provide Sample Collection Kits with a specific kit ID number, 
Pathoseal and sample transfer bag, pre-labelled blood and urine tubes, a Sample Information 
Sheet, GlycoScore Consent Form, to be completed for each donor.  The Sample Collection Kits 
will be transferred to LUB for processing and archiving. Whole blood will be collected into the 
pre-labelled K2 EDTA vacutainer/Paxgene blood tubes and processed as follows;8,9 

 
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7.6.1 GlycoScore Study Blood Samples (One EDTA tube collected for 
Medtechtomarket)  
For the first 5 samples, Medtechtomarket will be notified and fresh blood samples will be 
collected from LUB on the same day and transported back to Medtechtomarket. 
Medtechtomarket will process and run a fresh versus frozen comparison study.  It is 
anticipated that following the comparison study all remaining blood samples will then be 
processed by Liverpool University Biobank and stored frozen for collection by 
Medtechtomarket once a suitable number of samples have been collected.  The blood 
samples will be processed using the following sample procedure (either at Medtechtomarket 
or Liverpool Biobank):  
 
• After collection, the blood will be gently mixed by inverting the tube 8 to 10 times  
 
• The pre-labelled tubes will be stored at 2-8ºC   
 
• Liverpool biobank / Medtechtomarket will be notified, and the samples will be collected 
and processed the same day 
 
• The blood sample will be centrifuged for 10 minutes at 2,000 x g using a refrigerated 
centrifuge (samples should be centrifuged on the same day as sample collection) 
 
• The plasma layer will be carefully collected with an appropriate transfer pipette  
 
• Aliquots (0.2ml) of plasma will be pipetted into appropriately sized pre-labelled cryovials. 
This process will be completed within 1 hour of centrifugation 
 
• Aliquots will be stored in a specimen box or rack in a -80ºC freezer  
 
• Plasma samples will be tested once sufficient samples have been collected 
 
 
7.6.2 Liverpool University Biobank Urine Sample (storage at LUB for future 
research) 
• After collection, half of the urine sample will be decanted and given to the research team 
 
• The urine will then be decanted into the pre-labelled urine tube supplied in the Sample 
Collection Kit by the research nurse 
 
Samples will be collected by a member of the LUB team for processing and storage.  
 
7.6.3 Liverpool University Biobank Blood Sample (one EDTA and one Paxgene 
tube storage at LUB for future research) 
• After collection, the blood will be gently mixed by inverting the tube 8 to 10 times.  
 
• The pre-labelled tubes will be stored in the sample bag provided by LUB until collection.  
 

 
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• Samples will be collected by the Liverpool University Biobank Team who will spin down 
the samples and aliquot the plasma into 1ml cryovials, alongside the cell pellet, prior to 
freezing at -80°C. 
 
• The Paxgene blood sample will be processed according to Biobank site procedures 
 
Plasma samples will be destroyed at the end of the study unless the participant has indicated 
they would be happy for samples to be used in future ethically approved research, in which case 
the remaining samples will be stored at Medtechtomarket laboratories and the additional blood 
samples at the Liverpool University Biobank alongside their urine samples. 
 
 
7.7 WITHDRAWAL OF PARTICIPANTS 
Participants will be advised that they have the right to withdraw from the study at any time 
without prejudice, and may be withdrawn at the Investigator’s discretion at any time. In the event 
that a patient drops out of the study or is withdrawn, the withdrawal section of the 
eCRF/database should be completed. The Investigator should record the date of the withdrawal, 
who initiated the withdrawal, and the reason for the withdrawal (if the patient provided a reason) 
within the additional information.  The Investigator should inform Medtechtomarket of the 
participant study number of any withdrawn patients. Any data obtained from a withdrawn 
participant will be excluded from the study and any stored plasma or urine samples will be 
destroyed. Additional participants will be recruited to replace any participants who withdraw from 
the study. 
 
8 
STUDY OPERATOR SELECTION AND TRAINING 
Study Operators will be Medtechtomarket laboratory staff trained in how to run the PSA test (Dialab 
PSA ELISA Z00338) and the GlycoScore immunoassays.  All testing will be carried out at 
Medtechtomarket Laboratories. A minimum of 2 study operators will be required to conduct the 
study to ensure data is available from several operators.   
 
9 
STUDY PROCEDURES 
9.1 PARTICIPANT RECRUITMENT 
Potential participants will be sent a letter about the study with their biopsy appointment letter. 
Potential participants will be identified by the NHS staff member looking after the patients 
attending for their biopsy. Potential participants will then be introduced to the research/study 
nurse to discuss the study and whether they wish to participate. They will also be provided with a 
patient information sheet which explains the study. Potential participants will be asked if they are 
willing to donate three tubes of venous blood, a urine sample and asked permission for utilisation 
of archived biopsy tissue samples for future research purposes. A nurse associated with the 
study and trained to obtain informed consent for research projects will be available to answer any 
questions a potential participant might have and will be present to administer and witness 
consent before blood is taken.  A copy of the signed and dated written informed consent form 
and any other written information will be provided to the participant prior to donating (see Section 
18 for further details).  Bloods will be taken by the research nurse or by clinic staff trained in 
phlebotomy. 
Patients that meet the eligibility criteria will be enrolled onto the study. 

 
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The following information shall be recorded for all patients on the eCRF/database:  
 
• Demographics, to include race, ethnicity, age. 
• Dietary preference (required for patients consenting for future research) 
• Concomitant medications and dosages 
• Date of PSA test (if possible) and result 
• Date of mpMRI, mpMRI report and Likert or PI-RADS® score (if available) 
• Gleason grade and score (when available) 
• CPG grade (if available) 
• Additional relevant scan information, e.g. Bone Scan, CT scan or PSMA scan. 
 
9.2 SAMPLE COLLECTION 
Following informed consent, three tubes of venous blood will be collected from the participant. 
The procedure for collecting and processing of the plasma samples is detailed in Section 7.6.  
Once sufficient plasma samples have been received by Medtechtomarket they will be tested on 
the GlycoScore immunoassays and the PSA test (Dialab PSA ELISA Z00338).  
 
If consent for sample use in future research has taken place for the urine and biopsy tissue 
samples, the sample of urine obtained from the clinic, pre-biopsy and pre-dipstick testing will be 
transferred to LUB with the additional blood samples for processing and storage. The biopsy 
tissue sample will be processed according to standard procedures following the biopsy and 
stored in the pathology lab.  
A flowchart summarising each step of the study procedures can be seen in Appendix A. 
 
 
9.3  QUALITY CONTROL PROCEDURES 
A 5-point standard curve will be run with each biomarker immunoassay test according to the 
Instructions for Use. 3 replicates per sample will be run.  Data will be analysed for outliers using 
Grubbs test for quantitative data. All outliers will be investigated to determine the cause. The 
investigation and its conclusions will be documented in the report. 
 
 
9.4 PSA TEST QUALITY CONTROL PROCEDURES 
Quality controls will be run as per the Instructions for Use accompanying the kit. 
 
10 STUDY MONITORING 
Medtechtomarket Consulting Ltd will monitor the trial in accordance with ICH-GCP guidelines, ISO 
20916:2019 and applicable regulatory requirements, and ensure that trial initiation, conduct, and 
closure are adequate. The Investigator and site staff are expected to cooperate with 
Medtechtomarket Consulting Ltd personnel or agents of Medtechtomarket Consulting Ltd and to be 
available during the monitoring visits to answer questions and to provide any missing information. 
The Investigator(s)/institution(s) will permit direct access to source data/documents for trial-related 
source data verification, audits, Ethics Committee review, and regulatory inspection(s).  
 
 A detailed study monitoring plan will be provided separately to this study protocol. 

 
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10.1  SOURCE DATA VERIFICATION 
Source data verification (SDV) ensures accuracy and credibility of the data obtained. During 
monitoring visits, reported data will be reviewed with regard to being accurate, complete and 
verifiable from source documents (e. g., electronic clinical databases/applications including case 
notes (PENS), blood / pathology / scan results (ICE results server) alongside clinic letters 
(EPRO) and GP referral forms (available via eXchange). All data reported on the eCRF/database 
should be supported by source documents. 
 
10.2  DEFINITION OF SOURCE DATA  
Source data includes all information in source documents (original records and certified copies of 
original records, data, laboratory records) and includes all clinical findings or observations 
relevant to the study – as outlined within this protocol.  In this study the following information will 
be supplied on the eCRF/database and therefore may require verifying from source documents: 
 
• 
Diagnosis (or otherwise) of prostate cancer 
• 
Hospital derived PSA test results (recorded on referral) 
• 
mpMRI report and Likert or PI-RADS® score 
• 
Gleason grade and score 
 
10.3  AUDITING 
As a quality assurance measure, Medtechtomarket Consulting Ltd or regulatory authorities may 
undertake audits before, during and after the trial. Regulatory authorities and representatives of 
the relevant Ethics Committee will be allowed to conduct inspections at the site. The Investigator 
will notify Medtechtomarket Consulting Ltd if regulatory authorities contact them to schedule an 
inspection. 
 
11 DATA MANAGEMENT 
11.1 DATA HANDLING 
The Investigator is responsible for prompt reporting of accurate, complete and legible data in the 
eCRF/database and in all reports. Whenever possible, clinical data will be entered directly into an 
eCRF/database held locally on the host trust site. Pseudonymised data will be transferred to 
Medtechtomarket with appropriate linkage to the study sample ID. The eCRF/database is 
designed to provide a fully paperless solution to data entry, but will permit printed versions of the 
eCRF in real time, in a readable format, should a paper archive be required or necessary, The 
Investigator should maintain a list of personnel authorised to enter data into the eCRF.  The 
Investigator will keep a list containing all participants approached and enrolled into the trial. This 
list remains with the Investigator and is used for unambiguous identification of each subject. The 
list contains the participants’ study ID, full name, date of birth, date of enrolment in the trial, and 
the hospital number and National Health Service number, if applicable. 
 
The subject’s consent and enrolment in the trial must be recorded in the participant’s medical 
record. This data should identify the trial and document the dates of the participant’s trial 
participation.  
 
11.2 CASE REPORT FORM 
CRF data will be entered onto a bespoke backend password protected MS Access database 
which allows data to be entered into clearly defined portals.  The data is backed up by the Trust 

 
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on a twice-daily basis.  Screening CRF data which will hold the patient identifiable details is 
stored on a separate database. The eCRF/database will contain the following information: 
• 
The study participants’ identification number 
• 
Most recent PSA measurement obtained 
• 
Details of any current medication the patient is taking including medication name and 
dosage 
• 
Dietary preference (required for patients consenting for future research)  
• 
mpMRI result (Likert or PI-RADS® score) 
• 
Gleason grade 
• 
Gleason score 
• 
Diagnosis/outcome 
 
11.3 RECORD KEEPING 
To comply with international regulations, the Investigator will arrange for the retention of essential 
trial documents for a minimum period of 10 years after the end of the study.  
 
The Investigator should take measures to prevent accidental or premature destruction of records. 
If archiving can no longer be maintained at the site, the Investigator should notify 
Medtechtomarket Consulting Ltd. 
 
Data from the eCRF/database will be exported to Medtechtomarket which will contain all of the 
clinical information (see Section 11.2) using the participant study ID to link to the sample. 
participant's name and any other identifying detail will NOT be included in any study data 
electronic file. Study data from the electronic file will be manually inputted into a clinical data 
management system by double data entry cross-checking for accuracy. Post verification, the 
database will be locked. 
 
12 STATISTICAL CONSIDERATIONS 
12.1 DATA QUALITY CONTROL  
Direct entry of data into a clinical database will be the preferred mode of data entry. All data 
accepted as valid will be included.  
 
All procedural errors, instrument malfunctions and control failures will be investigated to 
determine the cause. The investigation and its conclusions will be documented in the report. 
 
Data will be analysed for outliers using Grubbs Test. All outliers will be investigated to determine 
the cause. The investigation and its conclusions will be documented in the report. 
 
12.2 STATISTICAL ANALYSIS OF PRIMARY OBJECTIVE  
The diagnostic performance of the individual biomarkers will be analysed by the calculation and 
comparison of the sensitivity and specificity, likelihood ratio, PPV and NPV and ROC analysis 
along with confidence intervals and significance levels.  Analysis will be carried out for: 
 
Each biomarker in combination with PSA  

 
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Each possible combination of biomarker with and without PSA 
 
12.3 STATISTICAL ANALYSIS OF SECONDARY OBJECTIVES 
Graphical plots and correlation statistics will be used to assess the correlation between the 
‘GlycoScore’ and Gleason score, Gleason grade and CPG grade.  
 
12.4 DETERMINATION OF SAMPLE SIZE 
12.4.1 Sample Size Calculations  
Calculations were performed in order to calculate the sample numbers required to achieve 
a statistical power of 80% for the primary objective of evaluating the diagnostic 
performance of the GlycoScore biomarkers for detection of clinically significant prostate 
cancer (Gleason score of ≥7). The main measures of diagnostic performance used in the 
power calculations were sensitivity and specificity. 
 
The reference method for diagnosis of prostate cancer will be biopsy, and the reference 
method for identifying patients who do not have cancer will be by a combination of mpMRI 
and biopsy.  
 
A pilot study involving two cohorts (York & Exeter) showed the following performance 
characteristics:  
Statistic 
Training set 
Test set 
Sensitivity 
85% 
67% 
Sensitivity confidence interval 
6% 
25% 
Specificity 
67% 
100% 
Specificity confidence interval 
10% 
28% 
 
From this pilot data, the following sensitivity and specificity estimates have been proposed 
for the calculation of sample size required to power a prospective study at 80%:  
 
  
Estimated Performance 
Statistic  
Estimate 
95% CI (lower) 
95% CI (upper) 
Sensitivity  
0.8 
 0.65 
0.95 
Specificity  
0.8 
 0.65 
0.95 
 
The study population has been confirmed to contain 356 clinically significant cases, and 
430 non-clinically significant/PCa negative controls, giving a disease prevalence of 45.3%. 
 
Sample size calculations were undertaken in R (version 4.1.2) using the MKmisc package 
(version 1.8) with formulae derived from Machin10 and Flahault et al11, and the previously 
outlined target values for sensitivity, specificity, and prevalence, along with listed 
confidence intervals, to derive the following sample size guidelines: 
 
Sample Size for Sensitivity Optimised cut-offs 
alpha 
0.05 Level of significance (one-sided) 
1-beta 
0.8 
Power 
Diseased patient cohort 
size 
62 
Estimated minimum number of subjects with 
disease condition for sensitivity 
Subjects without disease 
75 
Estimated minimum number of subjects without 
disease condition for sensitivity 

 
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Sample size for SN 
137 
Total sample size for confirming sensitivity  
Total subjects (N) 
137 
Recommended minimum sample size  
 
 
 
 
Sample Size for Specificity Optimised cut-offs 
alpha 
0.05 Level of significance (one-sided) 
1-beta 
0.8 
Power 
Diseased patient cohort 
size 
52 
Estimated minimum number of subjects with 
disease condition for specificity 
Subjects without disease 
62 
Estimated minimum number of subjects without 
disease condition for specificity 
Sample size for SP 
114 
Total sample size for confirming specificity 
Total subjects (N) 
137 
Recommended minimum sample size  
A minimum of 62 cases are required to provide 80% power at a 1-sided significance level 
of 0.05, and at a prevalence of 45.3%, it is predicted a minimum of 137 subjects in total 
would be required. 
 
Statistical analysis of secondary objectives will be performed but not necessarily with a 
defined statistical power. 
 
12.5 PROCEDURE FOR ACCOUNTING FOR MISSING, UNUSED OR SPURIOUS DATA 
All missing, unused and spurious data will be documented in the study report with an 
explanation.  
 
Data from participants who withdraw or drop out of the study will be excluded from the study and 
replaced.  
 
Only complete data sets (PSA, mpMRI and/or biopsy results) will be used in the data analysis.  
 
 
13 STUDY PROTOCOL AMENDMENTS 
Protocol changes may affect the legal and ethical status of the study and may also affect the 
statistical evaluations of sample size and the likelihood of the study fulfilling its primary objective. 
Protocol amendment(s) are distributed to Investigators with instructions on how to implement them.  
 
All protocol amendments, except non-substantial amendments i.e., logistical/administrative 
implications (e.g., change of monitor(s), change of telephone number) are submitted to the ethics 
committee for review and approval prior to implementation.  
 
 
14 DEVIATIONS FROM THE CLINICAL STUDY PROTOCOL 
The Investigator will not implement any deviation from, or changes of, the protocol without 
agreement by the Medtechtomarket Consulting Ltd and prior review and documented 
approval/favourable opinion from the ethics committee of an amendment, except where necessary 
to protect study participants rights safety and well-being, or the scientific integrity of the clinical 
performance study. 

 
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In the event that an emergency or other medical event occurs that requires a protocol deviation, the 
Investigator or designee will inform Medtechtomarket Consulting Ltd as soon as is possible. The 
protocol deviation will be documented in the eCRF/database. 
Any deviation from the protocol will be recorded and assessed as minor or major by 
Medtechtomarket Consulting Ltd.  For repeated and/or major deviations corrective and preventative 
actions will be developed by the site and Medtechtomarket which will be implemented promptly. 
It is the responsibility of the site to use continuous vigilance to identify and report protocol 
deviations. 
Medtechtomarket Consulting Ltd has the right to discontinue the study at any time.  In the event of 
premature discontinuation of the study, all study materials must be returned to Medtechtomarket 
Consulting Ltd.  
15 STATEMENTS OF CONFORMITY 
This protocol will be conducted under the relevant ICH guidelines commonly known as Good 
Clinical Practices, the applicable national requirements, and ethical principles that have their origins 
in the Declaration of Helsinki. 
 
The work described in this protocol will be undertaken following Research and Ethics Committee 
review. The study will not be initiated until written and dated full approval is obtained. 
 
16 INSURANCE 
Insurance coverage will be handled according to local requirements. 
Finance, insurance, and publication rights are addressed in separate agreements, as appropriate. 
 
17 ADVERSE EVENTS 
17.1  ADVERSE EVENT DEFINITION AND CATEGORISATION 
The GlycoScore immunoassays will be run at Medtechtomarket laboratories therefore there will 
be no risk to patients or site study site staff of adverse device effects or device deficiencies.   
 
The term adverse event (AE) describes events related to study activities. It refers to undesirable, 
negative events such as injuries or diseases observed over the course of the study.  
 
Participants in this study are subject to risks that are no greater than, or are similar to the risks 
associated with undergoing a blood draw from a peripheral vein. There is a very small risk of 
infection with standard peripheral venepuncture. To minimize the risk, standard precautions will 
be taken and where possible additional blood tubes will be added to a routine blood draw as part 
of the standard procedure for investigating suspected prostate cancer. Since blood sampling is a 
normal part of routine patient care, participating in this study should not cause any emotional 
stress or additional risks to either the study staff or participants.   
 
Urine collection is part of the pre-biopsy checks and not considered to pose any risk to the 
participants.  Obtaining biopsy tissue samples is the purpose of the transperineal biopsy 
procedure.  Patients are only consenting to their archived biopsy tissue samples being utilised for 
future research purposes and therefore there is no further to risk to the patient other than those 
outlined during consent for the biopsy procedure. 

 
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17.2 REVIEWING AND REPORTING ADVERSE EVENTS  
For this study the duration of patient participation is one visit therefore adverse event reporting 
would only be relevant once the patient has been registered onto the study and during 
blood/urine collection. Any adverse events that could result in a serious adverse event should be 
recorded on the SAE forms held in the Study Site File. Adverse events must be reviewed and 
categorised by the Investigator as soon as possible. Any serious adverse events (SAE) should 
be reported by email to the Medtechtomarket immediately, if possible, but no later than 3 
calendar days after site study staff awareness of the event. Any serious adverse events should 
be fully investigated with the findings recorded in a written report. 
 
SAE must be reported to the REC in accordance with the clinical site’s Research Governance 
policy and also reported to the Research and Development Manager at Medtechtomarket by 
email using the contact details below: 
 
 
Hannah Martin  
hannah.martin@medtechtomarket.com 
 
18 INFORMED CONSENT PROCESS 
A participant's informed consent must be obtained and documented in accordance with local 
regulations, ICH-GCP requirements, and the ethical principles that have their origin in the 
Declaration of Helsinki.  
 
Prior to obtaining informed consent, a complete explanation (both verbally and in written format) of 
the nature and the purpose of the study will be provided to the participant in a language and at a 
level of complexity understandable to them.  This shall take place under conditions where the 
participant has adequate time to consider the risks associated with their participation in the study.  
 
Prior to participation in the study the participant and the person who conducted the informed 
consent discussion must sign and date the Ethics Committee approved informed consent form. As 
part of the consent process, each subject must consent to direct access to their medical records for 
study-related monitoring, auditing, Ethics Committee review and regulatory inspection.  The 
participant will receive a copy of the signed and dated informed consent form. 
 
18.1 VULNERABLE POPULATION - PATIENTS LACKING CAPACITY TO CONSENT 
Patients lacking capacity to consent will not be recruited onto the study. 
  
19 SUSPENSION OR PREMATURE TERMINATION OF THE STUDY 
A principal Investigator, ethics committee, or regulatory authority can suspend or prematurely 
terminate participation in a clinical performance study at the study sites for which they are 
responsible.  
 
Medtechtomarket may also suspend or discontinue the study at any time if, in the sponsor’s opinion, 
there is a significant safety concern, relevant technical problems with the device or when monitoring 
or auditing identifies serious or repeated deviations from the protocol.  
 

 
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A justification in writing should be supplied by the terminating party and Medtechtomarket is 
responsible for informing the ethics committee and regulatory authority where necessary.  All close-
out activities shall be performed on termination of the study. 
 
If there is suspicion of an unacceptable risk to participants during the study, recruitment can be 
suspended whilst the risk is being assessed. 
20 PUBLICATION AND COMMUNICATION POLICY 
Details of publication rights are addressed in a separate contract, as appropriate. 
 
21 
REFERENCES 
1. 
https://www.nice.org.uk/guidance/ng131. (15 December 2021)  
 
2. 
https://www.nice.org.uk/guidance/ng131/evidence/d-diagnosing-and-identifying-clinically-
significant-prostate-cancer-pdf-6779081777. Page 17 
 
3. 
https://www.cancerresearchuk.org/health-professional/cancer-statistics/statistics-by-cancer-
type/prostate-cancer/incidence 
 
4. 
https://www.nhs.uk/conditions/prostate-cancer/diagnosis/ 
 
5. 
Roddam AW, Duffy MJ, Hamdy FC, et al. Use of prostate-specific antigen (PSA) isoforms for 
the detection of prostate cancer in men with a PSA level of 2-10 ng/ml: systematic review 
and meta-analysis. Eur Urol. 2005;48:386–99. doi:10.1016/j.eururo.2005.04.015. discussion 
398–9. 
 
6. 
Catalona WJ, Hudson MA, Scardino PT, et al. Selection of optimal prostate specific antigen 
cutoffs for early detection of prostate cancer: receiver operating characteristic curves. J Urol. 
1994;152:2037–42. 
 
7. 
Johnston E, Pye H, Bonet-Carne E, Panagiotaki E, Patel D, Galazi M, Heavey S, Carmona 
L, Freeman A, Trevisan G, Allen C, Kirkham A, Burling K, Stevens N, Hawkes D, Emberton 
M, Moore C, Ahmed HU, Atkinson D, Rodriguez-Justo M, Ng T, Alexander D, Whitaker H, 
Punwani S. INNOVATE: A prospective cohort study combining serum and urinary 
biomarkers with novel diffusion-weighted magnetic resonance imaging for the prediction and 
characterization of prostate cancer. BMC Cancer. 2016 Oct 21;16(1):816. doi: 
10.1186/s12885-016-2856-2. PMID: 27769214; PMCID: PMC5073433. 
 
8. 
Henry, JB (1979) Clinical Diagnosis and Management by Laboratory Methods, Volume 1, 
W.B Saunders Company, Philadelphia, PA, p 60. 
 
9. 
J. Proteome Res. 2009, 8, 1, 113–117 Publication Date:December 12, 2008 
  
https://doi.org/10.1021/pr800545q - Supporting Info 
 
10. Machin D, Wiley I. Sample size tables for clinical studies [electronic resource] / David Machin 
... [et al.]. 3rd ed. Chichester: Chichester: Wiley-Blackwell; 2009.  
 
11. Flahault A, Cadilhac M, Thomas G. Sample size calculation should be performed for design 
accuracy in diagnostic test studies. Journal of Clinical Epidemiology. 2005;58(8):859-62. 

 
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22 APPENDICES 
22.1 APPENDIX A – STUDY PROCEDURES FLOWCHART  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
STEP 6 - Blood samples collected by Medtechtomarket 
from LUB on the day of blood draw (for 1st 5 samples 
only) then by arrangement between LUB and 
Medtechmarket for the remaining frozen samples. 
Medtechtomarket to test blood samples for PSA and 
biomarker concentration, samples with consent for future 
use will then be stored at -80C at Medtechtomarket 
 
STEP 7 - Once hospital results become available study site 
staff to record PSA, mpMRI result (Likert or PI-RADS® 
Score 1-5) and biopsy result (Gleason score & grade 
group), on eCRF/database and send pseudonymised data 
to Medtechtomarket 
SITE/INVESTIGATOR 
RESPONSIBILITY 
MEDTECHTOMARKET  
RESPONSIBILITY 
 
STEP 8 - Medtechtomarket to ensure all results have been 
recorded for each patient 
 
STEP 3 (Visit 1) Once eligibility confirmed, patient is 
invited to clinic room with research team member, given 
written and verbal information regarding study, and asked 
if they consent to take 3 tubes of venous blood and a urine 
sample, in addition to future utilisation of biopsy tissue, if 
so the Consent Form is signed and patient data is entered 
onto the eCRF/Access database.  If patient does not 
consent urine is disposed of and the biopsy samples will 
not be available for future use.  
STEP 2 (Visit 1) – Patient provides urine sample as part of 
routine pre-biopsy check. Half of the urine sample is 
decanted pre-dipstick testing and stored until consent 
given.  A clinic staff member invites the patient to join the 
study and informs the research team (research 
nurse/research fellow).  Research team checks if patient is 
eligible for inclusion (only UTI negative can be enrolled).  
 
Pre-visit 1 
STEP 1 – Patients will be sent a letter explaining the study 
with their biopsy appointment letter 
STEP 5 (Visit 1) - Arrange for blood and urine samples to 
be collected as follows: 
 
GlycoScore Study blood samples  
LUB to collect the blood samples for processing on day of 
blood draw 
Blood, and urine samples for future research 
LUB to collect for processing/storage 
  
 
STEP 4 (Visit 1) – Research nurse/clinic phlebotomist 
collects one EDTA tube of venous blood sample for 
Medtechtomarket and one EDTA + 1 Paxgene tube for 
LUB using the pre-labelled tubes from the sample 
collection kit.  The urine sample from consenting patients 
is decanted into the pre-labelled urine pot in the sample 
collection kit and the Sample Information Sheet 
completed.  Inform biopsy staff where consent has been 
given for samples to be used for future use 
 
 

 
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22.2 APPENDIX B – SCHEDULE OF EVENTS 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Procedures 
Total number of participant visits - 1 
Pre-visit 1 
Visit 1 
Send out introduction 
letter/information leaflet to 
all transperineal biopsy 
patients with their biopsy 
appointment letter 
✓ 
 
Informed consent (+ core 
questions) 
 
✓ 
Medical history (case note 
review) 
 
✓ 
Concomitant medication 
check (at screening) 
 
✓ 
Blood sample collection  
 
✓ 
Urine collection processing 
for Liverpool Biobank 
 
✓ 
Blood sample collection 
processing – 1 tube for 
GlycoScore study, 2 tubes for 
Liverpool Biobank (1 x EDTA, 
1 x Paxgene tube) 
 
✓ 
CRF/eCRF completion 
including data transfer and 
query resolution 
 
When results are 
available