EBF Focus Meeting, Brussels, March 2015

Robert Nelson, Ph.D. MRQA

Head of Bioanalytical Assay Laboratory

Transfer and cross-validation of a PK assay

across 3 different sites

(Switzerland, UK and USA)(Ligand Binding Assay)

NovImmune SA

Geneva, Switzerland

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 20152

NovImmune – Company Profile

� Focused on advancing targeted medicines that address the causes rather

than symptoms of disease

� Proprietary next-generation antibody drug platform

� Bench-to-bedside & pilot manufacturing capabilities

� Pipeline of differentiated antibody-based products

� Scientific excellence

� Ten patent families

� 60+ peer-review journal

publications

� 30+ collaborations with leading

academic institutes

� Roche/Genentech Alliance

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 20153

� What drives Biotechs to outsource assays

� Case study: transfer and cross-validation of a PK

assay at 3 sites

� Lessons learned & conclusions

Presentation Outline

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 20154

What drives Biotechs to outsource assays?

� Expertise

� Limited internal resources

� Regulatory compliance (e.g. GLP)

� Logistical challenges

� Clinical study requirements

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 20155

� Challenging patient population

� Rare and life threatening disease indication

� Low sample volume

� Rapid turnaroundof data

� Multi-site global clinical trial

� Sample analysis at the right time, at the right

place to satisfy data delivery requirements

Case study: cross- validation of a PK assay at 3 sitesClinical study requirements

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 20156

� Gyrolab

� Fast data turnaround capacity

� Suitable with low sample volume

� Microfluids sensitive to sample/reagent quality

� Bridging assay

� Sequential format

Case study: cross-validation of a PK assay at 3 sitesAssay format

Drug (therapeutic mAb)

Biotinylated mouse

anti-idiotype mAb

Alexa Fluor mouse

anti-idiotype mAb

Gyrolab CD 200 nL

(With streptavidin coated beads)

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 20157

Case study: cross-validation of a PK assay at 3 sitesCRO requirements for the study

Sample receipt

Sample analysis

Data QC check

Data delivery

<24 hours

� Gyrolab technology availability

� Expertise

� Location (EU + USA)

� Short data delivery turnaround time

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 20158

� Assay developed at NovImmune (CH)

� Transfer + Validation at CRO 1 (UK)

� Validation at NovImmune (CH)

� Transfer + Validation at CRO 2 (USA)

� Cross-Validation at 3 sites (CH, UK, USA)

⇒ Ruggedness assessment

Case study: cross-validation of a PK assay at 3 sitesAssay transfer + validation strategy

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 20159

Case study: Transfer of a PK assay to 3 sitesAssay development at NovImmune (CH)

� Critical parameters defined

� Critical reagents

� Matrix effect/minimum requireddilution

� Working range & QC levels

� Selectivity & dilutional linearity assessed

� Transfer to CRO 1 (UK)

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 201510

Case study: Transfer of a PK assay to 3 sitesTransfer and validation at CRO 1 (UK)

Challenges

� New Gyrolab system

� Microfluidics present different challenges to ELISA/MSD

� Time to become familiar with the system and special

requirements

� Assay optimisation/troubleshooting required

� NovImmune analyst visit to CRO 1

� Technical support from Gyros

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Validation results

From 62.50 to 8000.00

Overall Intra- and Inter- accuracy and precision within criteria*

Normal matrices

Lipaemic and

Haemolysed matrices

Up to 1 in 2000 (in addition to 1 in 3 MRD)

No hook effect at 500 µg/mL

Room temperature 24 hours

Freeze/Thaw Cycle 5 cycles

18 months stability at both -20°C and -80°C

100% of naïve individual <LLOQ

Between 80% to 100% of individuals at the LoQC and HiQC

levels within criteria*

System suitability

Matrix Effect

Dilutional Linearity

Hook effect

Short term stability

Long Term Stability

Parameter

Working range of the method (ng/mL)

* %RE ≤± 20% and %CV ≤ 20%, except for LLOQ and ULOQ for which %RE ≤± 25% and %CV ≤ 25%,

Case study: Transfer of a PK assay to 3 sitesValidation summary

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BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 201512

Case study: Transfer of a PK assay to 3 sites

Validation completed successfully

� Transfer to CRO 2 (USA)

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 201513

Case study: Transfer of a PK assay to 3 sitesTransfer and validation at CRO 2 (USA)

Challenges

� New Gyrolab system

� Microfluidics present different challenges to ELISA/MSD

� Time to become familiar with the system and special

requirements

� Assay troubleshooting required

� NovImmune analyst visit to CRO 2

� Technical support from Gyros

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 201514

Case study: Transfer of a PK assay to 3 sites

Validation completed successfully

� Cross-validation at 3 sites

� NovImmune, Switzerland

� CRO 1,UK

� CRO 2, USA

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 201515

Transition

Ruggedness assessmentObjective and principle

� Establish inter-laboratory reliability

NovImmune

(Reference site)

CRO

in the UK

CRO

in the USA

Prepare and test 30 samples

Samples shipped

blinded to the

CRO sites

Testing at the CRO sites

Reference result

Observed result Observed result

Within criteria?

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� Analysis at NovImmune to set the reference concentration

� 2 assays

� Acceptance criteria:

� Assay valid (criteria met for calibration curve and QCs)

� Sample precision ≤ 20%

� Relative % difference between the 2 assays within ± 25%

Ruggedness assessmentSet up of the reference concentration

Mean Conc. from the two data sets = Reference concentration for other sites

≠ Theoretical concentration

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1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

Ruggedness assessmentNovImmune data

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

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� Shipped blinded and analysed at CRO site

Ruggedness assessmentShipment to the CRO sites

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Sample 2

Sample 3

…

…

Sample 29

Sample 30

Sample 25

Sample 3

Sample 16

…

…

Sample 1

Sample 28con

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n

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� Results calculated as follows:

� Target criteria:

Ruggedness assessmentData analysis

Relative Percentage Difference (%) = ((B-A)/A)*100

A = Reference result (Mean result from NovImmune)

B = Observed result (at the CRO site)

CV ≤ 20% and relative percentage difference within ± 25% of the reference

value for at least 80% of the samples tested

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Ruggedness assessmentData - 10 samples

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(Reference result)

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Ruggedness assessmentData – Relative percentage difference

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

87% of the samples within criteria ruggedness assessment successfully passed

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 201522

Lessons learned

� State expectations clearly from the start

� Kick off meetings to get to know the teams on both sides

� Ideally face-to-face meetings

� Share the background to get buy-in to the project

� Share “know-how” and provide support and advice

� Let the scientists talk to each other, share ideas and

troubleshoot

� Be open to client analyst working onsite at CRO

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 201523

Lessons learned

� Good communication at all stages of the relationship

� Expectations and timelines

� Problems and possible solutions

� Regular follow-up (TC, email, phone calls)

� Honest feedback from both sides

� What worked well

� What could be done better at CRO and Client side

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 201524

Conclusion

� Worked with CRO partners to meet challenging clinical

study requirements

� Assay validated at 3 different geographical locations

� Successfully supported data delivery for clinical trial at

the different sites

BOARD MEETING MARCH 2012EBF Focus Meeting, Brussels, March 201525

Contact information:

Robert Nelson

rnelson@novimmune.com

Thank you for your attention!