Appendix 1: Laboratory guidelines version 3.0 dd 07 August 2015
Guidelines for the collection of blood samples at different time points to monitor asparaginase
- Timepoints
Serum samples will be collected at different time points of routine blood examinations during asparaginase treatment. In case of an allergic reaction, 1 additional sample will be collected.
PEG E. coli asparaginase*
D0:before (nadir)and one hour after (peak) each PEG E. coli asparaginase administration.
D7: 7 days after each PEG E. coli asparaginase administration.
D14: 14days after each PEG E. coli asparaginase administration (=in IA nadir of second.
Native Erwinia asparaginase
Nadir native Erwinia asparaginase activity is measured 2 (or3) days after native Erwinia asparaginase administration (often prior to next aparaginase application).
Native E. Coli asparaginase *
Nadir native E. coli asparaginase activity is measured 3 (or 4) days after native E. coli administration (often prior to next aparaginase application).
The collections are performed in all de novo and relapse ALL and non-hodgkin lymphoma patients. Detailed schedules on ideal monitoring timepoints for each risk group are added in the following pages (p3-p12).
* If patients are switched from PEG (or native) E.coli asparaginase to native Erwinia asparaginase asparaginase, corresponding time points are used (see schedule on following pages, p3-p12).
Monitoring of asparaginase therapy in childhood acute lymphoblastic leukemia and lymphoma1/13
Appendix 1: Laboratory guidelines version 3.0 dd 07 August 2015
- Collection, storage and transport of serum samples
- Take blood sample on the time pointsas indicated in the respective schemes (p3-p12).
- Cool the centrifuge at 4°C whenever known a sample will be taken.
- Collect at least 2 serum tubes of 4 ml blood each.
- Put the samplesimmediately on ice after collection of the blood.
- Centrifuge the sample preferably immediately. If not possible, this should be marked on appendix 3.
- Centrifuge the samplesas soon as possible for 10 minutes cooled at a temperature of 4°C at 2500g. If these conditions are not possible, a maximum temperature of 8°C and a power of 1300g is allowed.
- Pipet the serum out of the sample tubes.
- Divide the serum equally in 3 labeled cryotubes(labels and tubes provided by the UZ Gent team).Each label is provided in duplo. Stick the duplicate label on the corresponding location of appendix 2.
- Put the cryotubes on dry ice to transport it to the freezer. The samples should be stored at -80°C immediately. If this is not possible, a temporary storage at -20°C is allowed, again, mention this in appendix 2.
- Send the completed appendix 2 to . For the pick-up of samples on the next day, please notify us before 13h by email to .
- As soon as appendix 2 is mailed, shipment of the samples will be arranged by the UZ Gent team.
- An email will be send to acknowledge receipt of appendix 1 and pinpoint date and hour when pick-up of the samples will take place.
Remark :For the first asparaginase administrations in 1A, samples at D0 (low, high) and D7will be picked up together whereas all other samples will be picked up in real-time (1-2 working days after sending appendix 3).
- The courier will arrive to the place as agreed (cfr address list) with a box of dry ice to transfer the samples to UZ Gent.
- Results of the asparaginase activity will be available in 3-4 working days.
- Send the completed appendix 3 after each treatment block to .
Example: Schedules of monitoring timepoints in the EORTC 58081
VLR, AR1 PATIENTS
Phase 1A
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseTP1 = D0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir
Native Erwinia aspa 2
D3-D4
.. / .. / .... / Nadir
Native E. coli aspa 2
D4-D5
.. / .. / .... / Nadir
Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir
Native Erwinia aspa 5
D10-D11
.. / .. / .... / Nadir
Native E. coli aspa 4
D11-D12
.. / .. / .... / Nadir
Native Erwinia aspa 6
TP1 D14
.. / .. / ....
= TP 2 D0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa2 / Native E. coli aspa 5 / Native Erwinia aspa 7
+ Peak (1h after)
D16-D17
.. / .. / .... / Nadir
Native Erwinia aspa 8
D17-D18
.. / .. / .... / Nadir
Native E. coli aspa 6
D18-D19
.. / .. / .... / Nadir
Native Erwinia aspa 9
D21
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 7 / Native Erwinia aspa 10
D23-D24
.. / .. / .... / Nadir
Native Erwinia aspa 11
D24-D25
.. / .. / .... / Nadir
Native E. coli aspa 8
D25-D26
.. / .. / .... / Nadir
Native Erwinia aspa 12
D28
.. / .. / .... / Nadir / Nadir / Nadir
= minimally required samples for optimal asparaginase monitoring
+ = recommended samples for optimal asparaginase monitoring
VLR, AR1 PATIENTS
Phase 2A
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir
Native Erwinia aspa 2
D3-D4
.. / .. / .... / Nadir
Native E. coli aspa 2
D4-D5
.. / .. / .... / Nadir
Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir
Native Erwinia aspa 5
D10-D11
.. / .. / .... / Nadir
Native E. coli aspa 4
D11-D12
.. / .. / .... / Nadir
Native Erwinia aspa 6
D14
.. / .. / .... / Nadir / Nadir / Nadir
VLR, AR1 PATIENTS
TPA= sample taken as soon as possible after allergic reaction
- Type of last aspa: PEG E. coli/native E. coli/ Erwinia aspa
- Date of last aspa: .. / .. / ….
- Date of allergic reaction: .. / .. / ….
- Date of aspa sample: .. / .. / ….
= minimally required samples for optimal asparaginase monitoring
+ = recommended samples for optimal asparaginase monitoring
AR2 PATIENTS
Phase 1A/ 1A’
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseTP1 = D0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir
Native Erwinia aspa 2
D3-D4
.. / .. / .... / Nadir
Native E. coli aspa 2
D4-D5
.. / .. / .... / Nadir
Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir
Native Erwinia aspa 5
D10-D11
.. / .. / .... / Nadir
Native E. coli aspa 4
D11-D12
.. / .. / .... / Nadir
Native Erwinia aspa 6
TP1 D14
.. / .. / ....
= TP 2 D0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa2 / Native E. coli aspa 5 / Native Erwinia aspa 7
+ Peak (1h after)
D16-D17
.. / .. / .... / Nadir
Native Erwinia aspa 8
D17-D18
.. / .. / .... / Nadir
Native E. coli aspa 6
D18-D19
.. / .. / .... / Nadir
Native Erwinia aspa 9
D21
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 7 / Native Erwinia aspa 10
D23-D24
.. / .. / .... / Nadir
Native Erwinia aspa 11
D24-D25
.. / .. / .... / Nadir
Native E. coli aspa 8
D25-D26
.. / .. / .... / Nadir
Native Erwinia aspa 12
D28
.. / .. / .... / Nadir / Nadir / Nadir
= minimally required samples for optimal asparaginase monitoring
+ = recommended samples for optimal asparaginase monitoring
AR2 PATIENTS
Phase 2A
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir
Native Erwinia aspa 2
D3-D4
.. / .. / .... / Nadir
Native E. coli aspa 2
D4-D5
.. / .. / .... / Nadir
Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir
Native Erwinia aspa 5
D10-D11
.. / .. / .... / Nadir
Native E. coli aspa 4
D11-D12
.. / .. / .... / Nadir
Native Erwinia aspa 6
D14
.. / .. / .... / Nadir / Nadir / Nadir
AR2 PATIENTS
HD-MTX1 in maintenance
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2
.. / .. / .... / Nadir
Native Erwinia aspa 2
D7
.. / .. / .... / Nadir / Nadir / Nadir
D14
.. / .. / .... / Nadir / Nadir / Nadir
= minimally required samples for optimal asparaginase monitoring
+ = recommended samples for optimal asparaginase monitoring
AR2 PATIENTS
HD-MTX2 in maintenance
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2
.. / .. / .... / Nadir
Native Erwinia aspa 2
D7
.. / .. / .... / Nadir / Nadir / Nadir
D14
.. / .. / .... / Nadir / Nadir / Nadir
AR2 PATIENTS
HD-MTX3 in maintenance
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2
.. / .. / .... / Nadir
Native Erwinia aspa 2
D7
.. / .. / .... / Nadir / Nadir / Nadir
D14
.. / .. / .... / Nadir / Nadir / Nadir
AR2 PATIENTS
HD-MTX4 in maintenance
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2
.. / .. / .... / Nadir
Native Erwinia aspa 2
D7
.. / .. / .... / Nadir / Nadir / Nadir
D14
.. / .. / .... / Nadir / Nadir / Nadir
AR2 PATIENTS
HD-MTX5 in maintenance
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2
.. / .. / .... / Nadir
Native Erwinia aspa 2
D7
.. / .. / .... / Nadir / Nadir / Nadir
D14
.. / .. / .... / Nadir / Nadir / Nadir
AR2 PATIENTS
HD-MTX6 in maintenance
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2
.. / .. / .... / Nadir
Native Erwinia aspa 2
D7
.. / .. / .... / Nadir / Nadir / Nadir
D14
.. / .. / .... / Nadir / Nadir / Nadir
AR2 PATIENTS
TPA= sample taken as soon as possible after allergic reaction
- Type of last aspa: PEG E. coli/native E. coli/ Erwinia asparaginase
- Date of last aspa: .. / .. / ….
- Date of allergic reaction: .. / .. / ….
- Date of aspa sample: .. / .. / ….
= minimally required samples for optimal asparaginase monitoring
+ = recommended samples for optimal asparaginase monitoring
VHR PATIENTS
Phase 1A
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseTP1 = D0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir
Native Erwinia aspa 2
D3-D4
.. / .. / .... / Nadir
Native E. coli aspa 2
D4-D5
.. / .. / .... / Nadir
Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir
Native Erwinia aspa 5
D10-D11
.. / .. / .... / Nadir
Native E. coli aspa 4
D11-D12
.. / .. / .... / Nadir
Native Erwinia aspa 6
TP1 D14
.. / .. / ....
= TP 2 D0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa2 / Native E. coli aspa 5 / Native Erwinia aspa 7
+ Peak (1h after)
D16-D17
.. / .. / .... / Nadir
Native Erwinia aspa 8
D17-D18
.. / .. / .... / Nadir
Native E. coli aspa 6
D18-D19
.. / .. / .... / Nadir
Native Erwinia aspa 9
D21
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 7 / Native Erwinia aspa 10
D23-D24
.. / .. / .... / Nadir
Native Erwinia aspa 11
D24-D25
.. / .. / .... / Nadir
Native E. coli aspa 8
D25-D26
.. / .. / .... / Nadir
Native Erwinia aspa 12
D28
.. / .. / .... / Nadir / Nadir / Nadir
= minimally required samples for optimal asparaginase monitoring
+ = recommended samples for optimal asparaginase monitoring
VHR PATIENTS
Phase 1B augmented
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir / Nadir
Native E. coli aspa 2 / Native Erwinia aspa 2
D4-D5
.. / .. / .... / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 4 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir / Nadir
Native E. coli aspa 5 / Native Erwinia aspa 5
D11-D12
.. / .. / .... / Nadir / Nadir
Native E. coli aspa 6 / Native Erwinia aspa 6
D14
.. / .. / .... / Nadir / Nadir / Nadir
VHR PATIENTS
Phase VANDA
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir
Native Erwinia aspa 2
D3-D4
.. / .. / .... / Nadir
Native E. coli aspa 2
D4-D5
.. / .. / .... / Nadir
Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir
Native Erwinia aspa 5
D10-D11
.. / .. / .... / Nadir
Native E. coli aspa 4
D11-D12
.. / .. / .... / Nadir
Native Erwinia aspa 6
D14
.. / .. / .... / Nadir / Nadir / Nadir
VHR PATIENTS
Phase 2A
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir
Native Erwinia aspa 2
D3-D4
.. / .. / .... / Nadir
Native E. coli aspa 2
D4-D5
.. / .. / .... / Nadir
Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir
Native Erwinia aspa 5
D10-D11
.. / .. / .... / Nadir
Native E. coli aspa 4
D11-D12
.. / .. / .... / Nadir
Native Erwinia aspa 6
D14
.. / .. / .... / Nadir / Nadir / Nadir
VHR PATIENTS
Phase 2A bis
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginaseD0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir
Native Erwinia aspa 2
D3-D4
.. / .. / .... / Nadir
Native E. coli aspa 2
D4-D5
.. / .. / .... / Nadir
Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir
Native Erwinia aspa 5
D10-D11
.. / .. / .... / Nadir
Native E. coli aspa 4
D11-D12
.. / .. / .... / Nadir
Native Erwinia aspa 6
D14
.. / .. / .... / Nadir / Nadir / Nadir
VHR PATIENTS
TPA= sample taken as soon as possible after allergic reaction
- Type of last aspa: PEG E. coli/native E. coli/ Erwinia asparaginase
- Date of last aspa: .. / .. / ….
- Date of allergic reaction: .. / .. / ….
- Date of aspa sample: .. / .. / ….
= minimally required samples for optimal asparaginase monitoring
+ = recommended samples for optimal asparaginase monitoring
- For other protocols
Actual date / Pegylated E. coli asparaginase / Native E. coli asparaginase / Native Erwinia asparaginase
D0
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli-aspa1 / Native E. coli aspa 1 / Native Erwinia aspa 1
+ Peak (1h after)
D2-D3
.. / .. / .... / Nadir
Native Erwinia aspa 2
D3-D4
.. / .. / .... / Nadir
Native E. coli aspa 2
D4-D5
.. / .. / .... / Nadir
Native Erwinia aspa 3
D7
.. / .. / .... / Nadir / Nadir / Nadir
Native E. coli aspa 3 / Native Erwinia aspa 4
D9-D10
.. / .. / .... / Nadir
Native Erwinia aspa 5
D10-D11
.. / .. / .... / Nadir
Native E. coli aspa 4
D11-D12
.. / .. / .... / Nadir
Native Erwinia aspa 6
D14
.. / .. / .... / Nadir / Nadir / Nadir
PEG E. coli asparaginase *
D0:before (nadir) and one hour after (peak) each PEG E. coli asparaginase administration.
D7: 7 days after each PEG E. coli asparaginase administration.
D14: 14 days after each PEG E. coli asparaginase administration (=nadir of subsequent administration, if appropriate)
Native Erwinia asparaginase
Nadir native Erwinia asparaginase activity is measured 2 (or 3) days after native Erwinia asparaginase administration (often prior to next aparaginase application).
Native E. Coli asparaginase *
Nadir native E. coli asparaginase activity is measured 3 (or 4) days after native E. coli administration (often prior to next aparaginase application).
Monitoring of asparaginase therapy in childhood acute lymphoblastic leukemia and lymphoma1/13