1. JACOB Central Laboratory Procedures and Sample Management

2. Where tumor tissue and biosamples are sent
To Targos
Tumor tissue for HER2 testing to ascertain subject eligibility for study
(Ambient)
To Covance
All ‘on study’ mandatory and optional samples
(Frozen)

3. First Communication with Sites (Except Japan)
Confirmation of site’s contact details
Correct mistakes and enter any missing information
Fax the completed form back within 24 hours
Dry ice provider and courier information in Europe
Contact Covance CLS with changes

4. Initial Supplies Review packing list within initial shipment
Visit-specific collection kits
Visit
Cycle 1 Day 1, Cycle 2 Day 1
RCR
Tumor Tissue Kit
All Sites
5 kits each
3 kits
10 kits
To obtain highest quality specimens possible, store collection supplies at recommended 4ᵒC to 25ᵒC (39ᵒF to 77ᵒF)

5. Initial Supplies Covance CLS Investigator Manual
Provided in multiple languages
Requisition forms, Licenses, CVs, and Reference Ranges remain in English
In China, Requisitions are provided in Mandarin
Collection and packaging instructions
Laboratory certifications
Covance CLS contact information
Courier contact information
Provided by fax in Europe
Please review manual prior to first patient visit

6. Initial Supplies Quick Reference Guide Laminated Synopsis
Provided in most local languages
Kit information tips
Requisition completion tips
Resupply
Inventory control tips
Laminated Synopsis
Collection instructions per visit
Packaging
Material resupply
Covance CLS contact information

7. Initial Supplies Shipping supplies Bulk supplies
Shipping boxes (except Japan)
Site specific shipping documents
Air waybills, Commercial Invoices, Customs Declaration template, return address labels (except China and Japan)
Shipping supplies are provided by courier or Covance CLS, depending on courier arrangements
Refer to Covance CLS Investigator Manual and courier arrangements fax for country specific information
Bulk supplies
Positively charged slides

8. Ambient Specimens/Tumor Tissue Kit supplied by Covance and shipped to Targos (Germany)
Tumor Blocks
Tumor Slides (within 48 hours of sectioning)

9. Covance Initial Supplies: Tissue Collection Review packing list within initial shipment
HER2 screening collection kit:
Tumor Tissue Kit
All Sites
10 kits
To obtain highest quality specimens possible, store collection supplies at recommended 4ᵒC to 25ᵒC (39ᵒF to 77ᵒF)

10. Tissue Collection: Key Points (1/4)
Archival Tumor Specimens
Study sites should send patients’ tumor specimens to Targos as early as possible during screening period to avoid delay
Specimens must be FFPE tumor blocks (10% neutral buffered formalin)
If FFPE block (or partial block) unavailable because of country or site regulations, then slides may be accepted
Slides should only be submitted if no possibility of submitting blocks
Minimum 15 unstained slides requested for central review of HER2 status, MET expression, and exploratory biomarker research

11. Tissue Collection: Key Points (2/4)
Archival Tumor Specimens
Failure to follow these instructions could delay results reporting:
Fill out requisition form provided in tissue kit
De-identify associated pathology report
Black-out patient identifiers and replace with subject screening number
Ensure ID on path report and block match
Insert white copy of completed requisition form and copy of de-identified pathology report in outer pocket of specimen biohazard bag
Retain pink copy of requisition form and copy of pathology report in study binder
Write screening ID on specimen label on sample container
Ship samples at ambient temperature to Targos
HER2 testing results will be faxed to you within 3 working days of sample receipt at Targos (for samples delivered before noon)

12. Example of result report form

13. Tissue Collection: Key Points (3/4)
Archival Tumor Block
Ensure block is labelled with block ID
If block IDs not used, please record patient screening number on block
Place block in plastic bubble wrap padding cut to size
Place wrapped block in 60 mL plastic orange top container
Place container inside largest compartment of specimen biohazard bag and seal
Place specimen biohazard bag into ambient kit box and close
Block will be returned to site address on file unless you specify otherwise on Requisition form

14. Tissue Collection: Key Points (4/4)
Archival Tumor UNSTAINED Slides
Label slides in a solvent resistant manner (e.g. pencil or diamond etching) with study number BO25114, patient screening number and block ID (do not use adhesive labels)
Record date slides were sectioned on requisition form
Section the paraffin block as follows:
Sections should be 4 microns thick
Mount 15 serially-cut (consecutive) sections onto the SuperFrost charged slides provided in the tissue kit
SAMPLES SUBMITTED ON INCORRECT SLIDE TYPE WILL NOT BE TESTED!!!
Allow slides to dry completely
Place slides in slide container provided in tissue kit
Wrap slide container in bubble wrap and place inside the specimen biohazard bag and seal
Place specimen biohazard bag into ambient kit box and ship slides within 48 hours of sectioning

15. Tumor Tissue: Requisition Completion
Samples are submitted to Targos
Full page example

16. Covance: Kit Inventory Review
Kits are automatically resupplied, except:
Tissue kits
Lost, damaged or expired kits
Latin America, Russia, Thailand
Review for expired kits in preparation of patient visits
Use shorter expiration date first
Destroy expired kits or kits/supplies in excess at the end of study
Do not return to Covance CLS
Inform Covance CLS of lost, damaged, discarded or expired kits

17. Covance CLS Resupply Requests (1/2)
Request following supplies from Covance CLS
Replacement of damaged, lost or expired kits
All kits in Latin America, Russia, Thailand
Shipping supplies (except when provided by courier)
Tissue kits
Order supplies online:
Secure website
Confirmatory  sent providing work order reference number

18. Covance CLS Resupply Requests (2/2)
Allow minimum of 7-9 working days for the delivery of your resupply order
Exceptions:
12 in China
24 in Japan + additional days for customs clearance to receive your resupply order 18

19. Initial Supplies: Blood sample collections Review packing list within initial shipment
Visit-specific collection kits
Visit
Cycle 1 Day 1, Cycle 2 Day 1
RCR*
All Sites
5 kits each
3 kits
* Optional RCR consent required for collection
To obtain highest quality specimens possible, store collection supplies at recommended 4ᵒC to 25ᵒC (39ᵒF to 77ᵒF)

20. Blood Sample Collection Kit Content
All collection items for specified visit
Tubes pre-labelled with test names
Write Screening/Subject Number on tube labels
2 extra bar coded labels
If tube replacement required
Serum PCR
Serum PCR

21. Accession Number A unique identifier assigned to each kit
Do not interchange kits between sites or protocols!
Do not interchange tubes between kits!

22. Laboratory Requisitions
Completion is mandatory for sample processing
Provided in English (except China)
All information completed in English
Complete all required information and answer questions
Collection instructions and shipping condition
Ship white copies to Covance CLS with samples
Retain pink copies for your records

23. Requisition Completion Completion is mandatory for sample processing
METGASTRIC SHOW LUNCH TO END
23-Apr-17
Requisition Completion Completion is mandatory for sample processing
Subject Number 1
To identify your patient
Birthdate
Complete month field in English
(Example: 02 JUN 1968)
Day
Month
Year 1 J U L 1 9 8
Male
Female
Sex X
Collection Date
Complete month field in English
(Example: 01 JAN 2001)
Day
Month
Year 1 J A N 2 1 3
To calculate
specimen stability
24 Hours Clock
Collection Time
(Record Midnight as 23:59) 08 : 00
Requisition Completed by
Full name in capital letters
DR JOHN SMITH
To assist in site
communication
Samples Collected by
Full name in capital letters
DR JOHN SMITH
Phone Number
Clear + Consistent + Complete = Query-Free Requisitions

24. Requisition Completion Provide information using check boxes or written entries
Provide collection date (DD-MMM-YYYY) and collection time (24 hr) for each time point
Respond  Sample not collected if the sample was not collected
Incomplete information triggers queries

25. Frozen Specimens sent to Covance CLS
Frozen (ship day of collection or next day)
Serum
SM1 Trastuzumab PK Pre-Dose
SM2 Trastuzumab PK Post-Dose
SM3 Pertuzumab PK Pre-Dose
SM4 Pertuzumab PK Post-Dose
SM5 ATA Pertuzumab
SM6 Serum RCR
SM9 HER2 ECD/Ligands
Plasma
SM7 Plasma RCR
Whole Blood
SM8 Whole Blood RCR
SM = Specimen Management
Covance CLS forward samples to specified laboratory for analysis as directed by sponsor
Testing not performed and not reported by Covance CLS

26. Sample Collection Key Points
Follow detailed collection instructions provided
In Covance CLS Investigator Manual
On requisitions within the kits
On Laminated Synopsis
Ship samples day of collection or next day
Maintain at frozen until shipment
Good Quality DATA Begins with Good Quality Samples

27. Sample Collection Key Points
Before sample collection
Serum and plasma processing
Check tube’s expiry date
Centrifuge 15 minutes, 1500 – x g, 4ᵒC, within 60 minutes of blood collection (same conditions apply if no cooled centrifuge available)
Do not use expired tubes
Ensure you chose correct tubes labelled with correct sample name, patient and sample number
Transfer to corresponding transfer tube and invert to mix serum/plasma
Sample collection
Draw blood sample and invert times
Aliquot into one tube to obtain ~ 3 mL
Clot serum sample at room temperature for at least 30 minutes or until clotted
Aliquot within 20 min after centrifugation is finished

28. Sample Collection Key Points
Prepare specimens for shipment
Freeze preferably at -70ᵒC
If not possible, at -20ᵒC
Record sample processing, storage and shipping in Sample log
Discard used blood collection and transfer tubes according to local regulations
Ship to Covance as soon as possible after collection (either same day or next morning)
Special Instructions
RCR samples can only be taken if separate RCR informed consent has been given

29. Sample Collection Key Points
Sample tubes vary per specimen type
Collect
Invert
Clot Serum only
Centrifuge Except whole blood
Aliquot Except whole blood
Freeze
6 mL blood sample
8-10 times
Clot sample at room temperature for at least 30 minutes or until dotted
Serum only
15 minutes, 1500 – 2000 x g, 4ᵒC, within 60 minutes of blood collection (same conditions apply if no cooled centrifuge available)
Except whole blood
Transfer to 5 mL transfer tube and invert to homogenize the plasma. Then aliquot into one tube to obtain ~1.5 mL. Aliquot within 20 min after centrifugation is finished
Preferably at -70ᵒC, if not possible: at -20ᵒC =
15-25ᵒC
= 1 inversion

30. General Packaging Reminders
Store samples at appropriate temperature
Ambient
Frozen -70ᵒC, -20ᵒC
Use Covance CLS/courier provided boxes (except Japan)
In Japan, no shipping boxes are used
Plan ahead for pickup and dry ice requests
May require 48 hour advance notice
Cancelling your request
Consider holiday shipping schedules
Consolidate according to shipping temperature
One accession number per Specimen Collection Bag and per shipping temperature
Complete and send required shipping documents
Avoid delays

31. Shipping Documents (Except Japan)
Pre-Printed provided by Covance CLS or courier
Air waybills, Commercial Invoices and Customs Declaration template
Complete required information
Send/attach to appropriate shipping box
Ensure Commercial Invoice/Customs Declaration is included
Customs Declaration
Commercial Invoice

32. Examples of Queries 1 1 Subject Number
Missing, incomplete or inconsistent data on requisitions 1 1
Subject Number
Incorrect identifiers on tubes
Pat ID____
HER2 ECD
11 K-M
Multiple accessions used for 1 visit

33. Specimen Pending Arrival Report
Faxed to sites once per month
List samples that are:
> 30 days old
Any frozen samples cancelled as not received since last report
Samples are considered lost when 62 days have passed
Reminds sites to send samples to Covance CLS

34. Synergy with MetGastric study
JACOB study accepts HER2 test result from Targos and vice versa
In those MetGastric and JACOB overlapping sites (25%), screen for MetGastric study first
Where patient screened in MetGastric study has HER2 positive status:
Obtain Informed Consent for JACOB
Investigator/study nurse registers MetGastric study patient screen number in the JACOB’s IXRS screening module
Send results transfer form with detailed patient information to Targos
→ Tissue sample will be transferred within Targos
If site is not a JACOB/MetGastric overlapping site, a referral network is recommended

35. Jacob/MetGastric Sample Sharing
ICF signed for MetGastric
Screening Call in IXRS for MetGastric
Dispatch FFPE tumor block to Targos (or at least 15 slides)
HER2 testing performed at Targos
Continue with remaining screening activities until HER2 result from Targos is available
HER2-
Randomization call in IXRS
HER2+
Screen Failed Call in IXRS for MetGastric
ICF signed for JACOB
Screening for JACOB in IXRS; enter MetGastric screening number as reference
Results Transfer form completed by site and sent to Targos
Randomization for JACOB in IXRS

36. Recruitment Challenge: High Screen Failure Rate
It is assumed that screening ineligibility rate may be ~85%
Primarily driven by requirement for HER2 positivity
~ 22% of patients with advanced gastric cancer are HER2+
But… it’s not impossible: ToGA trial recruited successfully with a ~80% screen failure rate
Improve overall screening rates by implementing process of pre-identifying and pre-screening patients
This will help reduce screening failure rate not related to HER2 positivity
If local pre-screening HER2 result is borderline or equivocal, please still send for central HER2 testing
Reviewing pre-screening information will help identify why potentially eligible patients are not considered for screening