JACOB Central Laboratory Procedures and Sample Management.

JACOB CentralLaboratory Procedures

and Sample Management

2

Where tumor tissue and biosamples are sent

To TargosTumor 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 informationFax 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 ManualProvided in multiple languages• Requisition forms, Licenses, CVs, and

Reference Ranges remain in English• In China, Requisitions are provided in

Mandarin

Collection and packaging instructionsLaboratory certifications Covance CLS contact informationCourier contact information • Provided by fax in Europe

Please review manual prior to first patient visit

6

Quick Reference GuideProvided in most local languagesKit information tipsRequisition completion tipsResupplyInventory control tips

Laminated SynopsisProvided in most local languagesCollection instructions per visitPackaging Material resupplyCovance CLS contact information

Initial Supplies

7

Initial Supplies

Shipping suppliesShipping 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 suppliesPositively charged slides

8

Ambient Specimens/Tumor Tissue Kit supplied by Covance and shipped to Targos (Germany)

Ambient

Tissue Tumor Blocks Tumor Slides (within 48 hours of sectioning)

9

Covance Initial Supplies: Tissue CollectionReview 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 SpecimensStudy 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 kitDe-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)

1212

Example of result report form

13

Tissue Collection: Key Points (3/4)

Archival Tumor BlockEnsure 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 sizePlace wrapped block in 60 mL plastic orange top containerPlace container inside largest compartment of specimen biohazard bag and sealPlace 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 sealPlace specimen biohazard bag into ambient kit box and ship slides within 48 hours of sectioning

15

Tumor Tissue:Requisition Completion

Samples are submitted to TargosFull page example

16

Covance: Kit Inventory Review

Kits are automatically resupplied, except:

Tissue kitsLost, damaged or expired kitsLatin 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,

ThailandShipping supplies (except when provided by courier)Tissue kits

Order supplies online: www.covance.com/kitordering

Secure websiteConfirmatory e-mail sent providing work order reference number

18

Covance CLS Resupply Requests (2/2)

Exceptions:

12 in China

24 in Japan + additional days for customs clearanceto receive your resupply order

Allow minimum of 7-9 working days for the delivery of your resupply order

19

Initial Supplies: Blood sample collectionsReview 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

To obtain highest quality specimens possible,store collection supplies at recommended 4ᵒC to 25ᵒC

(39ᵒF to 77ᵒF)

* Optional RCR consent required for collection

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

1 2 3 4 5Serum PCR

1 2 3 4 5Serum PCR

21

Accession NumberA 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

To identify your patient

Phone Number

Subject Number

BirthdateComplete month field in English

(Example: 02 JUN 1968)

Sex

Collection DateComplete month field in English

(Example: 01 JAN 2001)

Collection Time(Record Midnight as 23:59)

Requisition Completed byFull name in capital letters

Samples Collected byFull name in capital letters

0 1 J U L 1 9 8 0

0 0 0 0 1

X

0 1 J A N 2 0 1 3

08 00

DR JOHN SMITH

DR JOHN SMITH

+44 0131 732 787

24 Hours Clock

:

Day Month Year

Male Female

Day Month Year

To calculate specimen stability

To assist in site communication

Clear + Consistent + Complete = Query-Free Requisitions

24

Incomplete information triggers queries

Requisition Completion Provide information using check boxes or written entries

Provide collection date (DD-MMM-YYYY) and collection time (24 hr) for each time pointRespond Sample not collected if the sample was not collected

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 ManagementCovance CLS forward samples to specified laboratory for analysis as directed by sponsorTesting 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 DATABegins with

Good Quality Samples

27

Sample Collection Key Points

Before sample collectionCheck tube’s expiry date• Do not use expired tubes

Ensure you chose correct tubes labelled with correct sample name, patient and sample number

Sample collectionDraw blood sample and invert 8-10 times• Clot serum sample at room

temperature for at least 30 minutes or until clotted

Serum and plasma processingCentrifuge 15 minutes, 1500 – 2000 x g, 4ᵒC, within 60 minutes of blood collection (same conditions apply if no cooled centrifuge available)Transfer to corresponding transfer tube and invert to mix serum/plasmaAliquot into one tube to obtain~ 3 mL• Aliquot within 20 min after

centrifugation is finished

28

Sample Collection Key Points

Prepare specimens for shipmentFreeze preferably at -70ᵒC• If not possible, at -20ᵒC

Record sample processing, storage and shipping in Sample logDiscard used blood collection and transfer tubes according to local regulationsShip to Covance as soon as possible after collection (either same day or next morning)

Special InstructionsRCR 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 onlyCentrifuge

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

finishedExcept whole blood

Preferably at -70ᵒC, if not possible: at -20ᵒC

= 1 inversion

15-25ᵒC

=

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 temperatureOne accession number per Specimen Collection Bag and per shipping temperature

Complete and send required shipping documents• Avoid delays

31

Commercial InvoiceCustoms Declaration

Shipping Documents (Except Japan)

Pre-Printed provided by Covance CLS or courierAir waybills, Commercial Invoices and Customs Declaration template

• Complete required information• Send/attach to appropriate shipping box• Ensure Commercial Invoice/Customs Declaration is included

32

Examples of Queries

Subject Number 1 1Missing, incomplete or

inconsistent data on requisitions

Incorrect identifiers on tubes

Pat ID____HER2 ECD

Multiple accessions used for 1 visit

11 K-M

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 SharingICF signed for MetGastric

Screening Call in IXRS for MetGastric

Dispatch FFPE tumor block to Targos (or at least 15 slides)

HER2 testing performed at TargosContinue 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