Impact of Sample Handling and Processing on Bioanalycial Outcome
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Transcript of Impact of Sample Handling and Processing on Bioanalycial Outcome
IMPACT OF SAMPLE HANDLING AND
PROCESSING ON BIOANALYTICAL
OUTCOME
Haiko Pillu
Head Technical Operations
CPU Antwerpen
SAFETY & EFFICACY CLINICAL TRIAL SOLUTIONS SGS Life Science Services Biopharm Day Seminar – Antwerp, October 29, 2015
2 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
INTRODUCTION
Data from clinical assays (biomarkers, PK, PD, and
immunogenicity) are often key outcomes from clinical trials
Implementing these endpoints in clinical trials is very :
Costly
time - and resource-consuming
Ensuring appropriate measures are taken from the sample
collection until the completion of laboratory testing is
paramount.
3 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
SAMPLES WITHIN CLINICAL TRIALS
Past Clinical studies sample
treatments
PK samples
Low amount of PD samples
Sample treatment
1 step handling
Amount of time per sample
= limited
Techniques
Centrifugation
Aliquoting
Freezing
Occasional sample preparations
with larger sample
treatments/procedures
Present Clinical studies larger/ expanded sample
treatments
PK assesments
PD assesments : more specific
treatments on sample preparation
Sample treatments
Multiple steps during treatments
sample treatments up to 5h for each
sample /batch of samples
Large amount of samples/aliquots
up to 10 different assays on 1 time
point
Specific conditions
Time consuming treatment schedules
New processes new techniques
Implementation new techniques
Need for training, specific qualifications
Running pilot studies
evolution
4 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
SAMPLES WITHIN CLINICAL TRIALS
Past Clinical studies sample
treatments
PK samples
Low amount of PD samples
Sample treatment
1 step handling
Amount of time per sample
= limited
Techniques
Centrifugation
Aliquoting
Freezing
Occasional sample preparations
with larger sample
treatments/procedures
Present Clinical studies larger/ expanded sample
treatments
PK assesments
PD assesments : more specific
treatments on sample preparation
Sample treatments
Multiple steps during treatments
sample treatments up to 5h for each
sample /batch of samples
Large amount of samples/aliquots
up to 10 different assays on 1 time
point
Specific conditions Time consuming treatment schedules
New processes new techniques
Implementation new techniques
Need for training, specific qualifications
Running pilot studies
evolution
5 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
SAMPLES WITHIN CLINICAL TRIALS
Lab manuals getting more specific/demanding
Specific conditions/requests
Timelines to centrifugation
Timelines after centrifugation
Timelines to storage
Storage conditions
Matrix used
Additional handling steps
….
All this makes it more challeging to maintain good
sample quality and to plan resources
6 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
WHAT’S ON THE OTHER END?
Why are all these parameters set?
How do we get to these (more complex) sample treatments?
What are the thoughts/reasoning behind it?
How is it proven to be effective?
How will information passed by from lab to site?
7 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
SETTING THE RIGHT
PARAMETERS
Source :” The effects of anticoagulant choice and sample
processing time on hematologic values of juvenile
whooping cranes” (Joan Maurer,Betsy Reichenberg, Cristin Kelly,Barry K. Hartup)
Case study describes collection blood and the outcome
with following factors
2 anti- Coagulants used (K3 EDTA and LiHE)
Slides made immediatly versus 4-6h delay
Questions
Does the anti coagulatant has an impact on results?
Does the processing time have an impact on results?
Is there any correlation of factors between anti-Coalgulant
and/or sample processing time?
1
8 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
SETTING THE RIGHT PARAMETERS
The total granulocyte concentration(heterophils and
eosinophils; H/E concentration) of each of the divided
samples
9 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
SETTING THE RIGHT PARAMETERS
The relative (%) leukocyte counts of each of the divided
samples
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SETTING THE RIGHT PARAMETERS
In this specific case study no effect has been seen on
results depending on your anti-coagulant if an immediate
sample processing was feasible.
However when having a time delay (sample processing) this
have an impact
Validation of techniques on specific parameters is key to
check if :
• goals are reached
• final results provided are fully “appropriate and correct” to
analysis demands and results
11 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
VALIDATION –
TIME & RESOURCE
Part 1 : Method Implementation and qualification
Set up and testing of PBMC protocol for preparation and
testing
Testing of PBMC stimulation procedure
Testing labelling procedures
Lysis, fixation, permeabilization buffer selection
Acquisition and analysis templates/gating strategy
Preliminary testing of method reproducibility
Bio
analy
tical Lab
1 m
onth
2
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Part 2 : In-vitro feasibility testing of the effect of
product on CD4 Th1/Th2 and Treg response
Set-up of culture conditions and stimulation (reagents,
stimulus, duration of culture, of stimulation)
Dose- and time effect of product (n= up to 6 subjects)
Test in-vitro effect on both CD4 Th1/ Th2 and Treg responses
Culture conditions in duplicate
FACS testing in duplicate
Data processing and statistical analysis
Bio
analy
tical Lab
1 m
onth
VALIDATION –
TIME & RESOURCE
13 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
Part 3 : Validation of a FACS method for analysis of
CD4 Th1/ Th2 subset (CD3, CD4, IFN-Gamma, IL-4) and
T Regulatory (CD3, CD4, CD25, FoxP3, CD127)
Sensitivity
Reproducibility:
• Between replicates;
• Between runs ;
• Between analysts;
• Between donors;
• Between two FACS systems.
Stability testing (e.g. storage of PBMC and effect of
cryopreservation, stability of reagents)
Robustness
Bio
analy
tical Lab
1 m
onth
VALIDATION –
TIME & RESOURCE
14 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
Part 4 : transition of the method to the clinical site
Providing procedure
Providing training
• Handling steps
• Conditions
• Go’s don’t go’s
Running pilot study qualification staff
• Between replicates Analyst evaluation
• Between analysts method/training validation
Reviewing results
Running clinical trial
Clin
ical S
ite
1 m
onth
VALIDATION –
TIME & RESOURCE
15 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
VALIDATION –
TIME & RESOURCE
Part 5 : Testing of CD4 Th1/ Th2 subset (CD3, CD4,
IFN-g, IL-4) and T Regulatory (CD3, CD4, CD25, FoxP3,
CD127) in clinical study samples
PBMC stimulation / 230 samples
CD4 Th1/ Th2 (CD3, CD4, IFN-Gamma, IL-4) FACS analysis/
230 samples
T Regulatory (CD3, CD4, CD25, FoxP3, CD127) FACS
analysis/ 230 samples
Bio
analy
tical Lab
X m
onth
s
16 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
OTHER EXAMPLES
Urine collections : adding of additional products as Tween
or BSA to avoid interference on tubes
Determination of Cytokines : use of a non standard blood
collection tube such as Tru Culture tubes
Sputum induction: effect of using Sputolysin during
handling on end parameters
Use of matrix : effect of presence of binding factors on
specific compounds/ parameters
Storage conditions : use of snapfreezing samples, storage
at -20°C or -70°C
…
17 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
CONCLUSION
Choose wise on parameters such as
Sampling tubes (anti coagulant)
Conditions
Sample processing times
Methods
…..
Validation of techniques is crucial
Implementation time consuming
Communication/training between bioanalytical lab and site
is key to :
get good final results
understand potential pitt falls for bioanalytical outcome
18 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
HOW DO WE ASSIST YOU IN THIS PROCESS?
Validation and choosing the right method is key for good
results
How do we achieve a good technique to deliver very good
Data when running a clinical trial?
Specialists on the bioanalytical part, Set up/validation
techniques
Specialists on the site part excecution
Running pilot studies between site and the BAN lab
• Handling of samples?
• Conditions?
• Experience
• Communication flow
19 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
PHARMACOLOGY SYNERGIES
- FROM LAB TO CLINICAL (AND VICE VERSA) -
A Clinical Pharmacology Unit with GMP pharmacy
Class II GCP laboratory
Fahmp & mec agreement
Mass Spectrometry & Immunoassays Experts
4 GLP/GMP Bioanalytical Laboratories 25 years experience
700 methods validated
31 LC-MS/MS
Services for small and large molecule testing in TK, PK and PD Online clinical samples dosing with CPUs
Discovery biomarkers translated in clinical research
Immune function testing
• Immunogenicity, flow cytometry, cytokine multiplexed ELISA
Biopharmaceuticals - Cell characterization
Metabolite profiling and mass balance studies (14C-labelled drug)
20 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
Life Science Services Pillu Haiko
Head Technical Operations
SGS Belgium NV Phone: + 32 3 217 25 77
Clinical Pharmacology Unit Fax: +32 (0) 3 217 25 81
Lange Beeldekensstraat 267 E-mail : [email protected]
2060 Antwerpen
Belgium Web : www.sgs.com/lifescience
THANK YOU FOR YOUR ATTENTION
+ 41 22 739 9548
+ 1 866 SGS 5003
+ 65 637 90 111
+ 33 1 53 78 18 79
+ 1 877 677 2667
+ 33 1 41 24 87 87
21 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015
QUESTIONS ?
22 SGS LIFE SCIENCE SERVICES BIOPHARMA DAY – OCTOBER, 29 2015